Nimba (Azadirachta indica)

Recent Research on Nimba (Azadirachta indica)

Anti-Fertility Activity – 

• Neem oil has been reported to possess strong spermicidal action against rhesus monkey and human spermatozoa within 30 seconds of mixing with semen. (Sinha et al, 1984).
• The lethal concentration for humans, spermatozoa was 1000 mg and 250 mg per 100 in 5 and 30 minutes, respectively. (Sharma & Saxena, 1959).
• An injection of neem oil (1001) into the uterine horn created an immunological response and prevented pregnancy for five months. (Upadhyay et al, 1990)

Anti-Inflammatory & Anti-Arthritic Activity – 

• Nimbidin (40 mg/ kg body weight) significantly reduced acute paw edema in rats induced by phlogistic agents, carrageenin, and kaolin. Its action is compared to phenytoin (100 mg/ kg). (Pillai & Santha Kumar. 1981).
• Bark extract produced significant anti-inflammatory activity. (Tidjani et al; 1989, Lorenz, 1976 & Kakai, 1976).

Cardiovascular Activity – Extract of neem leaves showed profound hypotension and a minimal negative chronotropic effect. It also exhibited a weak antiarrhythmic activity in rabbits against oculain-induced dysrhythmia. (Thompson & Anderson, 1978).

Anti-Diabetic Activity – 

• Aqueous extract of leaves decreased the blood sugar levels in dogs (on i.v. admn.) and prevented adrenaline as well as glucose hyperglycaemia. The onset of action was at 30 min- 4 hours. (Murthy et al, 1958).
• Neem oil does not produce hypoglycemia in normal animals, but it does improve glucose tolerance in partially pancreatectomised dogs. The hypoglycemic effect is about 24.77 % to 26.38 % in five hours (Pillai & Santha Kumari, 1981).

Skin Disorders & Anti-Microbial Activity – 

• It is found to be effective in case of acute eczema, ringworm infestation, and scabies (Singh et al, 1978). 
• The antimicrobial activity of different neem extracts and neem products is well documented. (Shankara Narayanan, 1965; Rao et al, 1986; Singh et al, 1988; Patel & Trivedi, 1962).

CNS Activity –

Leaf extract showed varying degrees of CNS depressant activity in mice. (Singh et al; 1987; Singh et al; 1980)

Immunostimulant Activity –

• The aqueous extract of bark showed anti-complementary activity, acting both on alternative and classical pathway activation (Vander et al, 1987 & 1989).
• The aqueous extract of the leaf possessed potent immunostimulant activity as evidenced by both humoral and cell-mediated responses (Sen et al, 1992).

• Kumar, Sanjay & Vandana, Udaya & Agrawal, divya & Hansa, Jagadish. (2015). Analgesic, Anti-inflammatory, and Anti-Pyretic Effects of Azadirachta indica (Neem) Leaf Extract in Albino Rats. International Journal of Science and Research (IJSR). 4. 713- 721. Neem (Azadirachta indica), popularly known as ‘Wonder tree’, is an evergreen tropical tree, which has been reported to have beneficial effects on various disease pathologies from ancient times. Such health benefits are the sum of molecular actions of the natural ingredients present in it. Among all ingredients, azadirachtin, nimbidin, flavonoids, and triterpenoids are notable for their beneficial effects against disease pathology. Neem has been reported to have anti-allergenic, anti-dermatic, anti-feedant, anti-fungal, anti-inflammatory, anti-pyorrhoeic, and anti-scabic modes of action. Besides these, neem is also beneficial for cardiac, diuretic, insecticidal, larvicidal, nematicidal, spermicidal, and other biological activities. Though here in the present study, our focus was only on three parameters, i.e., whether neem leaf extracts (NLE) can contribute to anti-analgesic, anti-inflammatory, and antipyretic action in albino rats, which is a novel one. Our experimental observations revealed that NLE has analgesic, anti-inflammatory, and antipyretic effects in Albino rats.
• Baghel, Shalu & Singh, Charanjeet. (2019). ANTIHYPERLIPIDEMIC ACTIVITY OF METHANOLIC EXTRACT OF AZADIRACHTA INDICA. Journal of Biomedical and Pharmaceutical Research. 8. 10. 32553/ jbpr. v8i6. 678. Hyperlipidemia often results from delayed or defective clearance or overproduction of VLDL by the liver, which is subsequently transformed into LDL. The whole plant of Azadirachta indica was collected from the local areas of the Jaipur district of Rajasthan. The plant is authenticated by the University of Rajasthan by Dr. Manju Sharma, dept of Botany. Selected medicinal plants were cut into small pieces, cleaned, and shade-dried at room temperature, then subjected to physical evaluation with different parameters. Macerated 5 g of air-dried drug coarsely powdered with 100 ml of ethanol of the specified strength in a closed flask for twenty-four hours, shaking frequently during six hours, and allowed to stand for 18 h. About 2-3 g of powder is accurately weighed in a china dish and kept in a hot air oven maintained at 110±1 °C for four hours. After cooling in a desiccator, the loss in weight was recorded. This procedure was repeated till a constant weight was obtained. For insoluble ash value, boil the ash for 5- 10 minutes with 25 ml of diluted hydrochloric acid, collect the insoluble matter in a Gooch crucible, wash with hot water, ignite, and weigh. The whole plant of Azadirachta indica was reduced to fine powder and was subjected to maceration in a 1-liter conical flask for 7 days at room temperature with alcohol (70 %) and water (30 %). In the present study, we aimed to screen various extracts of selected plants on the cholesterol-induced hyperlipidemic rat model. Thus, the results of the present investigation clearly indicated that the selected medicinal plants possess good antihyperlipidemic activity in atherogenic diet-induced hyperlipidemic rats and led to the development of a new Herbal formulation possessing antihyperlipidemic and antiatherosclerotic activities. This is the first study that investigates hypolipidemic activity. 
• SALAZAR, Daniel & Hoyos, Rodrigo & Orozco-Sánchez, Fernando & Arango, Myrtha & Gomez Londoño, Luisa. (2015). Antifungal activity of neem (Azadirachta indica: Meliaceae) extracts against dermatophytes. Acta Biológica Colombiana. 20. 201- 207. 10. 15446/ abc. v20n3. 45225. To assess the antifungal activity of methanolic extracts from neem tree (Azadirachta indica A. Juss.), several bioassays were conducted following the M38-A2 broth microdilution method on 14 isolates of the dermatophytes Trichophyton mentagrophytes, Trichophyton rubrum, Microsporum canis, and Epidermophyton floccosum. Neem extracts were obtained through methanol-hexane partitioning of mature green leaves and seed oil. Furthermore, high-performance liquid chromatography (HPLC) analyses were carried out to relate the chemical profile to their content of terpenoids, widely known for their antifungal activity. The antimycotic Terbinafine served as a positive control. Results showed that there was total growth inhibition of the dermatophytes isolates at minimal inhibitory concentrations (MIC) between 50 μg/ mL and 200 μg/ mL for leaf extract, and between 625 μg/ mL and 2500 μg/ mL for seed oil extract. The MIC of positive control (Terbinafine) ranged between 0.0019 μg/ mL and 0.0313 μg/ mL. Both neem leaves and seed oil methanol extracts exhibited different chromatographic profiles by HPLC, which could explain the differences observed in their antifungal activity. This analysis revealed the possible presence of terpenoids in both extracts, which are known to have biological activity. The results of this research are a new report on the therapeutic potential of neem for the control of dermatophytosis.
• Mostofa, Mahbub & Choudhury, Mohammed & Hossain, Md.Monir & Islam, Md & Islam, Muh & Sumon, Md Mahmudul Hasan. (2008). Antidiabetic effects of Catharanthus roseus, Azadirachta indica, Allium sativum, and glimepiride in experimentally diabetic induced rats. Bangladesh Journal of Veterinary Medicine. 5. 10. 3329/ bjvm. v5i1.1324. Catharanthus roseus (Nyantara), Azadirachta indica (Neem), and Allium sativum (Garlic) are medicinal plants used in Ayurveda for treating various diseases, one of which is diabetes mellitus. In the present study of 12 12-month period from January to December 2007, aqueous extract of these plants was prepared, and the blood glucose-lowering effect and improvement of body weight gain in Streptozotocin (50 mg/ kg bwt i.p.) induced diabetic rats were measured and compared with that of the patent drug glimepride in the Department of Pharmacology, Bangladesh Agricultural University, Mymensingh. Rats were administered Catharanthus roseus, Azadirachta indica, and Allium sativum extracts at the dose rates of 1g/ kg, 500 mg/ kg, and 1g/ kg bwt orally for 14 days, respectively. Blood glucose level and body weight were measured by the Glucotrend kit and electronic balance, and compared with a patent drug, Glimepride, at a dose rate of 100 mg/ kg bwt. The data were compared statistically by using Student’s unpaired t-test. The herbal preparations of these plants significantly increased body weight gain and decreased blood glucose as compared with the patent drug. The present study clearly indicated the significant antidiabetic activity of Catharanthus Roseus, Azadirachta indica, and Allium sativum and supports the traditional usage of the herbal preparations by Ayurvedic physicians for the therapy of diabetics.
• Santra, Soumya & Naik, M.R. & Behera, R. & Agrawal, Divya & Kumar, Sanjay & Patnaik, S. (2014). Antipyretic effect of Azadirachta indica leaf extract (Neem Leaf Extract) on albino rats. Research Journal of Pharmaceutical, Biological and Chemical Sciences. 5. 669- 673. Evaluation of antipyretic activity of Neem Leaf Extract (NLE) by using Brewer’s yeast-induced pyrexia model. In this study, the albino rats were randomly divided into groups of six, each consisting of 6 rats. Group I: Control (distilled water 0.5 ml/ rat); Group II: Standard (Paracetamol 100 mg/ kg intraperitoneally); Group III, IV, V, VI (NLE 62.5, 125, 250, 500 mg/ kg body weight intraperitoneally, respectively). It is a randomized controlled experimental study. Fever was induced by injecting 20 ml/ kg body weight of 20 % suspension of brewer’s yeast subcutaneously below the nape of the neck. Those animals that had a constant rectal temperature or a variation of less than 1°C were included in the study. Drugs were given after the development of pyrexia, and temperatures were recorded. Paracetamol at 100 mg/ kg orally was taken as the standard drug. NLE 62.5 mg/ kg body weight showed no significant fall in temperature, whereas NLE 125 mg/ kg body weight decreased the temperature significantly from the 4th hour of its injection, along with NLE 250 mg and 500 mg/ kg body weight. NLE caused a significant fall in temperature from the 1st hour onwards. Distilled water did not show any antipyretic effect on Yeast-induced pyrexia. Thus,  NLE exhibited significant (p< 0.05) antipyretic activity with the progressive increase in dose.
• Raut, Ranjit R., & Sawant, Ajit, & Jamge, Bhagyshree. (2014). Antimicrobial activity of Azadirachta indica (Neem) against Pathogenic Microorganisms. Journal of Academia and Industrial Research. 3. 327- 329. Plants have been a source of herbal remedies throughout the history of mankind. Various medicinal plants have been used for years in daily life to treat diseases all over the world. In the present study, antimicrobial activity of Azadirachta indica was evaluated against Gram-negative pathogenic bacteria (Escherichia coli, Salmonella typhi, and Vibrio cholerae) and Gram-positive bacteria (Bacillus subtilis). Azadirachta indica leaves and barks were collected from the fields of Botanical Garden in Padmashri Vikhe Patil College, Pravaranagar, and pure cultures of the test organisms used for antimicrobial study were obtained from the Department of Microbiology, Pravara Institute of Medical Sciences, Loni. All the test organisms were screened for their antibacterial activity against the leaf and bark extract of A. indica by the agar well diffusion method. Leaf and bark extract of Azadirachta indica showed more inhibition zone against Vibrio cholerae and Bacillus subtilis, while E. coli and S. typhi are less susceptible to neem extract.
• Adamu, Muhammad & Lawan, Ibrahim & Abubakar, Ahmed & Dangora, Ibrahim. (2019). Antimicrobial Activity of Azadirachta indica (Neem) Leaf Extract Against Some Clinical Isolates. The present study was carried out at the Microbiology and Chemistry Laboratories, Federal University Dutse, Jigawa State. To investigate the activity of neem leaf extract against some clinical isolates. The test organism includes Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus. The phytochemical compounds present in the neem were extracted using ethanol as a solvent. The Soxhlet extractor was used to get the extract. Phytochemical screening was carried out using the standard methods. Some of the examined phytochemicals are Alkaloids, saponins, phenols, Tannins, and Flavonoids. The extract is more effective against Gram-negative E. coli and K. pneumoniae as compared to S. aureus, a Gram-positive bacterium. The zone of inhibition of the extract at 125 mg/ ml is 21 and 14. For Gram-negative and Gram-positive bacteria, respectively. The mean of the minimum inhibitory concentration of A. indica leaf extract, the minimum inhibitory concentration (MIC) of the extract in vitro revealed to be at much lower concentration 50mg/ml against E. coli as compared to 100mg/ml against S. aureus the minimum bactericidal concentration (MBC) of A. indica leaf extract shows that the extract have a bacteriostatic effect against gram negative bacteria with static effect against gram positive bacteria.
• Prabagar, Subramaniam & Nandakumar, Jeyarani & Prabagar, Jasotha & Thuraisingam, Suganja. (2020). SPC Journal of Plant Sciences Antimicrobial activity of Azadirachta Indica (neem) leaves and stem bark aqueous extracts. Background: Azadirachta indica is a multipurpose tree called Neem, which is used in Ayurvedic medicine for skin infections and as a household pesticide.  Methods: Aqueous extraction of plant material, such as leaf and stem bark, was investigated for anti-microbial activity without altering concentration.  Results: The Results of the study revealed that all extracts had inhibitory activity against Staphylococcus aureus with a clear zone of 2.6 cm, which is higher than the streptomycin control. The inhibition zone was found in descending order against Klebsiella, Pseudomonas, and Bacillus species. Antifungal activity of bark extract of Azadirachta indica against Aspergillus and Yeast showed the inhibitory growth of 64.2 % and 64 % whereas the leaf extract showed less inhibition than the bark extract.  Conclusion: The findings reveal that bark and leaf aqueous extracts have wide-ranging activity.
• Patil, Prabhakar & Patil, Sudha & Mane, Abhay & Verma, Sushilkumar. (2013). Antidiabetic activity of alcoholic extract of Neem Azadirachta Indica root bark. National Journal of Physiology, Pharmacy and Pharmacology. 3. 142- 146. 10. 5455/ njppp. 2013. 3. 134- 138. Background: Most of the parts of the neem tree were studied in detail by many researchers, but the neem root bark was not screened for antidiabetic effect. To evaluate the antidiabetic effect of neem root bark, this study was conducted. Aims & Objective: The present study was undertaken to evaluate the 70% alcoholic neem root bark extract (NRE) in diabetes. Materials and Methods: Basal blood sugar levels were estimated in overnight fasted Wistar albino rats of either sex. To evaluate the antihyperglycemic action of Neem root bark extract (NRE), an OGTT was done by giving glucose orally 60 minutes after giving the standard drug (Glibenclamide),  the Test drug (NRE) in the doses of 200, 400, and 800 mg/ kg, and blood sugar levels were estimated every half hour up to 4 hours. To evaluate hypoglycemic activity in alloxan-induced diabetic rats, similar doses were administered once daily for 15 days. Blood sugar levels were estimated by using a glucometer. Results: For the glucose tolerance test, Glibenclamide showed a significant (p< 0.01) reduction in blood sugar levels. The NRE showed statistically significant results in only the 800 mg/ kg dose. In comparison to glibenclamide, it did not show. In alloxan-induced diabetes, Glibenclamide showed a significant (p< 0.01) reduction in blood sugar levels. The NRE showed statistically significant results in only the 800 mg/ kg dose. Conclusion: Neem root has antihyperglycemic and hypoglycemic activity. It is not as significant as glibenclamide.
• Khan, Shaista. (2015). In Vitro Analysis of Antiallergic Activity of Neem (Azadirachta Indica) for Reduction of Wheat Allergens. Sindh Univ. Res. Jour. (Sci. Ser..). 47. 485- 488. Abstract: Plants are the origin of life on earth and have been essential for people’s livelihood since ancient times. Plants have been an excellent source of medicine. Survival of mankind is unfeasible without the plant kingdom. The name of the Neem tree in Sanskrit is ‘Arishtha’, which means that it is a reliever of ‘Sickness’. According to the healing system of Ayurveda, the Neem tree will strengthen individual resistance to disease, and Neem efficiently regulates the immune system and other functions. Neem is used for a wide range of disorders, including diarrhea, inflammatory diseases, antimicrobials, skin diseases, urinary disorders, antiseptics, bronchitis, cough, asthma, fever, hypertension, and infected burn skin diseases, malaria, and many more ailments. In the present study, characterization of wheat protein has been done, and possible allergens having molecular weights of 7 kDa, 9 kDa, 15 kDa, 12 kDa, 17 kDa, 36 kDa,, 45 kDa, and 77 kDa have been found. But after the treatment of wheat protein with Neem (Azadirachta indica) extract showed a high reduction of allergenic proteins. There are no allergic bands that have been seen after treatment with Neem. 
• Mankala, Dr. Santhosh Kumar. (2011). In vivo Antidiabetic evaluation of Neem leaf extract in alloxan-induced rats. Journal of Applied Pharmaceutical Science. 1. 100-105. Azadirachta indica has been used medicinally throughout history by many different cultures. Many compounds have been found in the exudates of the Azadirachta indica plant that have been used medically by humans. We have examined the pharmacological hypoglycemic action of Azadirachta indica in diabetic rats. After treatment for 24 hrs, Azadirachta indica 250 mg/ kg (single dose study) reduced glucose (18 %), cholesterol (15 %), triglycerides (32 %), urea (13 %), creatinine (23 %), and lipids (15 %). Multiple dose study for 15days also reduced creatinine, urea, lipids, triglycerides, and glucose. In a glucose tolerance test in diabetic rats with neem extract 250 mg/kg, glucose levels were significantly less compared to the control group. Azadirachta indica significantly reduces glucose levels on the 15th day in diabetic rats. Azadirachta indica serves as an important alternative source in the management of diabetes mellitus, involved in reducing increased blood glucose during diabetes, which should be examined further by oral hypoglycemic therapy.
• Thiagarajan, Kalaivani & Meignanam, E. & Premkumar, N. & Siva, Ramamoorthy & Vijayakumar, Vijayaparthasarathi & Chandrasekaran, Rajasekaran & Subramanian, Ramya & Jayakumararaj, R. (2009). Studies on Hepatoprotective Properties of Leaf Extracts of Azadirachta indica A. Juss (Meliaceae). Ethnobotanical Leaflets. 13. 165- 170. The present study was carried out to evaluate the hepatoprotective role of leaf extracts of Azadirachta indica A. Juss. Hepatoprotective activities of ethanolic and aqueous extracts of A. indica were examined against carbon tetrachloride-induced liver damage in mice using silymarin as a control. Enzyme activities of Serum Glutamate Oxaloacetate Transaminase (SGOT), Serum Glutamate Pyruvate Transaminase (SGPT), and Alkaline Phosphatase (ALP) were analyzed. Phytochemical leaf extracts of A. indica exhibited significant hepatoprotective activity. Ethanolic and aqueous leaf extracts of A. indica exhibited moderate activity over carbon tetrachloride-treated animals. Results confirm the traditional ethnomedicinal use of A. indica as a potential source of hepatoprotective agents.
• Chattopadhyay, Rabiranjan & Bandyopadhyay, M. (2006). Effect of Azadirachta indica leaf extract on serum lipid profile changes in normal and streptozotocin-induced diabetic rats. African Journal of Biomedical Research. 8. 10. 4314/ ajbr. v8i2. 35769. Effects of Azadirachta indica leaf extract on serum lipid profile changes in normal and streptozotocin-induced diabetic rats have been studied with a view to elucidating its possible effect on cardiovascular disease induced by hyperglycemia. It was observed that A. indica leaf extract significantly reduced the total cholesterol, LDL- and VLDL-cholesterol, triglycerides, and total lipids of serum in streptozotocin-induced diabetic rats, but HDL-cholesterol levels remained unchanged when compared with streptozotocin-induced diabetic control animals. African Journal of Biomedical Research Vol. 8 (2) 2005: 101- 104
• Shahbaz, Benish & Akhtar, Shabana & Muddassir, Maria & Yasmin, Shagufta & Akram, Ayesha & Majeed, Muhammad & Ahmad, Mukhtiar & Shafiq, Muhammad & Arshad, Shafia & Hadi, Faheem & Tahir Maqbool, Dr & Hayat, Sikander & Muhammad, Tahir. (2023). Anti-diabetic activity of a novel Phytoformulation of Tribulus terrestris, Prunus dulcis, Cicer arietinum, and Azadirachta indica in streptozotocin-induced diabetic rats. 19. 360- 373. Background and Objective: Selected medicinal plants have been investigated as an alternative source for the treatment of Streptozotocin (Stz) induced diabetes. The aim of the current study was to assess the anti-diabetic effects of selected plant formulations, Tribulus terrestris, Prunus dulcis, Cicer arietinum, and Azadirachta indica in diabetic rats induced by streptozotocin. Plants included in this phytoformulation have shown antioxidant, anti-diabetic, and anti-inflammatory properties evident in previous research.
• Basha, Saleem & Baruah, Mriganka & Shaker, A. & Kondaveeti, Dr.Suresh & Narayana, S. (2012). Hypoglycemic and hypolipidemic effect of Azadirachta indica seed oil and mehani (polyherbal formulation) on alloxan-induced diabetic albino rats. Research Journal of Pharmaceutical, Biological and Chemical Sciences. 3. 239- 246. The present study aims to investigate the hypoglycaemic and hypolipidemic effect of Azadirachta indica seed oil and Mehani (Poly Herbal Formulation) in alloxan-induced diabetic Wistar albino rats. The rats were divided into 6 groups, each consisting of 6 rats: Control (Normal) rats, diabetic induced rats received alloxan (150/ Kg in 3 doses), diabetic induced rats treated with Azadirachta indica seed oil (5ml/ Kg orally for 21 days), Normal rats treated with Azadirachta indica seed oil, Diabetic induced rats treated with Mehani (2 gm/ 60 Kg orally for 21 days) and Normal rats treated with Mehani. The rats were monitored for plasma glucose, glucose metabolising enzymes, hexokinase and glucose-6-phosphatase, and Lipid profile. Alloxan induced diabetic rats shows a high blood glucose with altered lipid profile and altered level of glucose metabolising enzymes which was counteracted upon treatment with Azadirachta indica seed oil and Mehani which shows significant (p< 0.01) reduction in blood glucose, serum cholesterol, serum triglyceride and glucose- 6- phosphatase level and significant (p< 0.01) increase in the level of hexokinase and serum HDL suggesting its hypoglycaemic and hypolipidemic role. Again, in comparison to Azadirachta indica seed oil, Mehani (Poly Herbal Formulation) has more hypoglycaemic and hypolipidemic effects.
• Satyanarayana, K & Sravanthi, K & Shaker, I., Anand & Ponnulakshmi, R. (2015). Molecular approach to identify antidiabetic potential of Azadirachta indica (neem) on the expression of insulin signaling molecules and glucose oxidation in skeletal muscle of high-fat and fructose-induced type-2 diabetic adult male rat. Journal of Ayurveda and Integrative Medicine. 10. 4103/ 0975- 9476. 157950. Background: Azadirachta indica (Neem) is a medicinal plant, used in Ayurveda for treating various diseases, one of which is diabetes mellitus. It is known to possess anti-inflammatory, antipyretic, antimicrobial, antidiabetic, and diverse pharmacological properties. However, the molecular mechanism underlying the effect of A. indica on insulin signal transduction and glucose homeostasis is obscure. Objective: The aim was to study the effects of A. indica aqueous leaf extract on the expression of insulin signaling molecules and glucose oxidation in the target tissue of high-fat and fructose-induced type-2 diabetic male rats. Materials and methods: The oral effective dose of A. indica leaf extract (400 mg/ kg body weight [b. wt]) was given once daily for 30 days to high-fat diet-induced diabetic rats. At the end of the experimental period, fasting blood glucose, oral glucose tolerance, serum lipid profile, and the levels of insulin signaling molecules, glycogen, and glucose oxidation in the gastrocnemius muscle were assessed. Results: Diabetic rats showed impaired glucose tolerance and impairment in insulin signaling molecules (insulin receptor, insulin receptor substrate- 1, phospho- IRS- 1(Tyr- 632), phospho- IRS- 1(Ser- 636), phospho- Akt (Ser- 473), and glucose transporter 4 [GLUT- 4] proteins), glycogen concentration and glucose oxidation. The treatment with A. indica leaf extract normalized the altered levels of blood glucose, serum insulin, lipid profile, and insulin signaling molecules as well as GLUT-4 proteins at 400 mg/ kg b.  wt dose. Conclusion: It is concluded from the present study that A. indica may play a significant role in the management of type-2 diabetes mellitus by improving the insulin signaling molecules and glucose utilization in the skeletal muscle.
• Awah, Francis & Uzoegwu, Peter & Ifeonu, Patrick. (2011). In vitro anti-HIV and immunomodulatory potentials of Azadirachta indica (Meliaceae) leaf extract. African Journal of Pharmacy and Pharmacology. 5. 1353- 1359. Azadirachta indica (neem) is attributed to have many ethnopharmacological properties and has been postulated to have a significant effect on CD4 counts in HIV/ AIDS subjects; however, the precise anti-retroviral mechanism is not yet known. In this study, a hydroacetone extract of A. indica leaves was investigated for its anti-retroviral and immunomodulatory potentials. The effect of the neem leaf extract on acute HIV infectivity and fusion was measured by the syncytia formation assay on HIV-1-infected C8166 CD4+ cells. The inhibitory effect of the extract on reverse transcriptase (RT) polymerase activity and HIV p24 antigen expression was evaluated in the culture supernatant. The extract was also assessed for its ability to decrease the phenotypic expression of the immune activation markers CD38 and CD69. Results from a syncytium formation assay indicated that the extract blocked HIV-1 envelope-mediated membrane fusion. The extract inhibited HIV- 1 replication in C8166 CD4+ cells in vitro by inhibiting the biochemical activity of HIV-1 reverse transcriptase, the result being a subsequent decrease in HIV p24 antigen concentration. In the effective dose range, no cytotoxicity was detected on uninfected target cells. Ex vivo, the extract exhibited a dose-dependent reduction in the levels of the immune activation marker CD38 and CD69 on phytohemagglutinin A (PHA)- stimulated human peripheral blood mononuclear cells (PBMC). The observed anti-HIV activity and immunomodulatory potentials of the extract show that neem could impart health benefits to HIV/AIDS patients, possibly by acting as fusion or reverse transcriptase inhibitors and also down-regulate hyperimmune activation.
• Chattopadhyay, Rabiranjan. (2004). Possible mechanism of hepatoprotective activity of Azadirachta indica leaf extract: Part II. Journal of Ethnopharmacology. 89. 217- 9. 10. 1016/ j. jep. 2003. 08. 006. Hepatoprotective activity of Azadirachta indica leaf extract against paracetamol-induced hepatic damage in rats has already been reported. In the present investigation, the effects of Azadirachta indica leaf extract on blood and liver glutathione, Na+ K(+) -ATPase activity, and thiobarbituric acid reactive substances against paracetamol-induced hepatic damage in rats have been studied with a view to elucidating the possible mechanism behind its hepatoprotective action. It was interesting to observe that Azadirachta indica leaf extract has reversal effects on the levels of the above-mentioned parameters in paracetamol hepatotoxicity. Possible mechanisms behind the results are discussed.
• Nagesh, Simhadri & M, Muniappan & I, Kannan & Subramanian, Viswanathan. (2018). ANTIFUNGAL ACTIVITY OF A SECONDARY METABOLITE OF AZADIRACHTA INDICA AND ITS DERIVATIVES – AN IN-SILICO STUDY. Asian Journal of Pharmaceutical and Clinical Research. 11. 175. 10. 22159/ ajpcr. 2017. v11i1. 22171. Objective: This study aimed to inhibit the 1, 3 β-glucan synthase with azadirachtin or with its derivatives by the docking method.Methods: The homology model of the protein 1, 3 β-glucan synthase was prepared with “easy modeler” using the query sequence and template, and it was validated with procheck of Ramachandran plot. The ligand was selected from the PubChem database, and the .sdf file was downloaded, which was converted to another file format with Open Babel. The files of protein and ligand were uploaded for rough docking with iGEMDOCK, and finally, the accurate docking was made with Autodock Vina. The docked poses were visualized with PYMOL and then saved. The derivatives of the ligand were generated with SWISS ADME, a free online software, and the derivative was selected for docking. Results: The results obtained from iGEMDOCK and Autodock Vina were tabulated. It was found out that the Azadirachtin and the derivatives are effective in binding 1, 3 β-glucan synthase and thereby inhibiting the formation and integrity of fungal cell wall. Conclusion: In this study, the secondary metabolite Azadirachtin and its derivatives show inhibitory action against the model protein 1 3 β-glucan synthase, and it was suggested that the external application of the ligand and its derivatives can be used because of their poor oral bioavailability.
• Maruthappan, K (2009). Hepatoprotective effect of Azadirachta indica (Neem) leaves against alcohol induced liver injury in albino rats. Journal of Pharmacy Research. Herbal drugs are traditionally used in various parts of the world to cure different diseases. The Ayurvedic and Siddha medical systems are very famous medical practices in Indian traditional medicine. The purpose of this study was to evaluate the hepatoprotective activity of A. indica using an ethyl alcohol induced liver injury in rats and probe into its mechanism of action. Liver damage was induced in Wistar rats (150- 200 g) by administering 30 % ethyl alcohol (1 ml/ 100 g b.wt.) once daily for 14 days. A. indica (1 g/ kg b. wt.) was given for 14 days. Silymarin (0.1 g/  kg b. wt.) was given as a reference drug once daily for 14 days. Levels of serum marker enzymes (AST, ALT, and ALP) were significantly (p< 0.05) increased in ethyl alcohol treated rats. Simultaneously, A. indica leaf powder and silymarin, a standard drug, significantly (p< 0.05) suppressed mainly the increase in plasma activities of AST, ALT, and ALP concentration, which are considered as markers of liver functional state. The significant (p< 0.05) decrease in the liver total protein (TP) and albumin (Alb) levels after ethyl alcohol treatment, which were reversed with A. indica. These effects were comparable to silymarin. To probe the possible mechanism by which A. indica prevents hepatic damage caused by ethyl alcohol, investigation on levels of thiobarbituric acid reactive substance (TBARS) was found to be elevated and significant (p< 0.05) decrease in glutathione reductase (GSH), superoxide dismutase (SOD) and catalase (CAT) content of liver after treatment with ethyl alcohol, which were significantly (p< 0.05) reversed by A. indica. The effects of A. indica were comparable to those of silymarin. The results of this study confirmed the hepatoprotective activity of the A. indica leaf powder.
• Baligar, Nagappa & Aladakatti, Ravindranath & Ahmed, Mukhtar & Hiremath, Murigendra. (2014). Hepatoprotective activity of the neem-based constituent azadirachtin-A in carbon tetrachloride intoxicated Wistar rats. Canadian journal of physiology and pharmacology. 92. 267- 77. 10. 1139/ cjpp- 2013- 0449. This study aimed to investigate the hepatoprotective role of azadirachtin-A in carbon tetrachloride (CCl4) induced hepatotoxicity in rats. The group allotment for the animals used in the hepatoprotective study included a vehicle treatment group, CCl4 (1 mL(kg body mass) (-1)) treatment group, silymarin (100 μg (kg body mass)(-1) day (-1)) + CCl4 treatment group, and groups treated with different doses of azadirachtin-A (100 or 200 μg  (kg body mass) (-1) day (-1)) + CCl4. On the 9th day, blood was obtained for measuring the biochemical parameters, and liver tissue was obtained for pathological examination. The acute toxicity test with azadirachtin-A (500, 1000, or 2000 μg (kg body mass) (-1)) indicated no mortality after 14 days of treatment; further, there was no change in behavior, food consumption, or organ mass. However, with the higher dose, some hematological parameters showed changes. Hepatoprotective studies revealed that the CCl4 treatment group exhibited a decrease in total protein and albumin levels, whereas a significant increase in BUN, AST, ALT, and ALP levels was noticed compared with the vehicle-treated control, indicating that there was liver damage caused by CCl4. Histology and ultrastructure study confirmed that pretreatment with azadirachtin-A dose-dependently reduced hepatocellular necrosis and, therefore, protected the liver against toxicity caused by CCl4. The results from this study indicate that pretreatment with azadirachtin-A at the higher dose levels moderately restores the rat liver to normal. This study confirms that azadirachtin-A possesses greater hepatoprotective action; however, the effective concentration needs to be determined.
• Sadasivam, Poompavai & Sree, v.gowri & RamSugitha, B. (2021). Enhanced Extraction and Improved Anticancer Activity of Neem ( Azadirachta indica ) Leaves by Electroporation Technique. IETE Journal of Research. 1-11. 10. 1080/ 03772063. 2021. 1962743. Plant-based medicines have been a part of primary healthcare since time immemorial. Natural products have received increasing attention to treat several regenerative diseases due to their availability and fewer side effects than conventional drugs. Towards this, Neem, commonly known as ‘Divine Life-giving tree’, has been investigated for its efficiency against MCF7, breast cancer cell lines. Ethanolic neem extracts were analysed for qualitative phytochemical screening tests, spectral analysis, antioxidant activity, and total phenol contents. In addition, to enhance the recovery of secondary metabolites from the plant extracts, the electroporation (EP) technique is used. The effect of EP was studied by comparing with untreated extracts, which exhibit antioxidant activity (∼96% increase) and total phenol contents (∼12.51 % increase) than untreated extracts. This indicates that electroporated extract (EE) exhibited higher antioxidant and phenolic activity than other extracts. Furthermore, different concentrations of untreated and electroporated neem extracts were used to study the anti-cancer activity. To enhance the uptake of extracts against cell membranes, electrical pulses of EP1 = 1200 V/cm, 100 s, 8 pulses and EP2 = 500 V/ cm, 20 ms, 8 pulses were used. The results indicate cell viabilities of 35 % with untreated extract only, 5.6 % with EE, 3.17 %, and 2.49 % with EP1 + EE extract and EP2 + EE extract, respectively, indicating the potential of the synergy of electrical pulses + EE. The viability was lowered by 5– 6 times after 72 h. This treatment is more appropriate to achieve maximum cell death, thereby reducing the side effects of conventional drugs in treating breast cancer.
• Nahak, Dr. Gayatri & Sahu, Rajani. (2010). Antioxidant activity in bark and roots of Neem (Azadirachta indica) and Mahaneem (Melia azedarach). Continental J. Pharmaceutical Sciences. 4. 28- 34. This study aimed to investigate the antioxidant activity of the multi-solvent extracts (aqueous, methanolic and ethanolic) of Root and Bark of two Medicinal plant i.e. Azadirachta indica A. Juss and Melia azedarach of Meliaceae family using 2, 2- diphenyl- 1- picryl- hydrazyl (DPPH) – scavenging assay. The dry powder of the Root and Bark of the two-neem tree was extracted using Soxhlet extraction followed by vacuum rotary evaporator methods. The extracts were tested for antioxidant activity using- DPPH-scavenging assay. Total phenol contents of extracts were also determined by the Folin-Ciocalteu reagent method. Experimental results revealed the highest fraction of crude extract, phenol content, as well as antioxidant activity in Barks and Roots of Mahaneem (Melia azedarach) in comparison to Neem (Azadirachta indica). The total phenolic concentration of the Bark and root of Melia showed a positive correlation with antioxidant capacity. Similarly, IC50 values in Bark and Root of Mahaneem were as low as those of Ascorbic acid. The plant Mahaneem and its plant parts may be exploited for clinical medicine as a potent factor because of their high antioxidant activity.
• Bharat, Pokhrel & Sagar, Rijal & Sulav, Raut & Pandeya, Ankit & eya. (2015). Investigations of antioxidant and antibacterial activity of leaf extracts of Azadirachta indica. African Journal of Biotechnology. 14.  3159- 3163. 10. 5897/ AJB2015. 14811. Active ingredients of medicinal plants have been used to cure several human diseases. Azadirachta indica is one of the most versatile medicinal plants, having a wide spectrum of biological activity due to the presence of a large number of bioactive compounds. The present study was conducted to evaluate antimicrobial and antioxidant efficiency and phytochemical screening of A. indica leaf extract using methanol as a solvent. A qualitative phytochemical screening was performed for the detection of various phytochemicals. Then, the quantitative determination of total phenols, flavonoids, and proanthocyanidins was done and expressed in terms of gallic acid and rutin equivalent. Total phenolic, flavonoid, and proanthocyanidin content were found to be 85.9± 4.0, 104.9± 5.5, and 65.4±  13.9 mg/g of plant extract, respectively. Also, the antibacterial activity was performed using six different bacterial strains: Escherichia coli (ATCC 25922), Staphylococcus aureus (ATCC 25923), Salmonella typhi (ATCC 14028), Klebsiella pneumoniae (ATCC 700603), Pseudomonas aeruginosa (ATCC 27853), and Proteus vulgaris (ATCC 35659). It was found that the maximum zone of inhibition of 22±3 mm was shown against S. aureus using 700 μg of plant extract. Similarly, the antioxidant activity of the methanolic extract of the plant was also determined, and it was found that the maximum inhibition obtained was 71.23 % when 500 μg of the plant extract was used. 
• Thakurta, Prarthana & DeBhowmik, Poulami & Mukherjee, Souryadeep & Hajra, Tapas & Patra, Amarendra & Bag, Prasanta K. (2007). Antibacterial, antisecretory, and antihemorrhagic activity of Azadirachta indica is used to treat cholera and diarrhea in India. Journal of Ethnopharmacology. 111. 607- 12. 10. 1016/ j. jep. 2007. 01. 022. Indigenous uses of Azadirachta indica A. juss (Maliaceae) (locally known as neem) leaves in different parts of India for curing gastrointestinal disorders such as diarrhea and cholera are widespread. The objective of the present study was to evaluate the antibacterial and antisecretory activity of neem extract against Vibrio cholerae, a causative agent of watery diarrhea, such as cholera. The methanol extract of neem leaf was tested for its antibacterial, antisecretory, and antihemorrhagic activity against Vibrio cholerae. Azadirachta indica extract had significant antibacterial activity against the multi-drug-resistant Vibrio cholerae of serotypes O1, O139, and non-O1, non-O139. The minimum inhibitory concentration reached by 50 % (MIC 50) and 90 % (MIC 90), and the minimum bactericidal concentration for the extract were 2.5, > 5, and 10 mg/ml, respectively. Neem extract showed antisecretory activity on Vibrio cholerae induced fluid secretion in the mouse intestine with inhibition values of 27.7 %, 41.1 %, 43.3 %, 57.0 %, and 77.9 % at doses of 100, 200, 300, 450, and 1800 mg/kg, respectively. Oral administration of the extract inhibited hemorrhage induced by Vibrio cholerae in the mouse intestine at a dose > of = 300 mg/ kg or greater. The results obtained in this study give some scientific support to the use of neem employed by the indigenous people in India for the treatment of diarrhea and the dreadful disease cholera.
• Pedroza, David & Benjamín, Serrano-Gallardo & Edith, Escobar-Ávila & Delia, Alma & Nava-Hernández, & Patricia, Martha & María, Vega-Menchaca & Carmen, Del. (2017). HIV-1 infection inhibition by neem (Azadirachta indica A. Juss.) leaf extracts and Azadirachtin. Indian journal of traditional knowledge. 16. 437- 441. Mexico is a country with a strong attachment to the use of traditional medicine, which is the heritage of pre-Hispanic ethnic groups of Mexican territory that are still present all across the country. The Mexican markets, dedicated exclusively to trade medicinal plants, have facilitated the use, getting, and importation of plants from other regions of the world, such as neem, which has been used for many health conditions, both non-communicable and infectious diseases, such as cancer, diabetes, bacterial, parasitic, and viral infections, even the infection with human immunodeficiency virus. , The objective of this study was to evaluate HIV-1 infection inhibition by neem leaf extracts and neem compounds. Cytotoxicity of aqueous and ethanolic neem leaf extracts and Azadirachtin and Limonene compounds was evaluated in CEM T-cells at concentrations of 1 ppm, 10 ppm (extracts); 1 µM, 10 µM (compounds). Only the aqueous extract concentration of 1 ppm and Azadirachtin at 1 µM and 10 µM concentrations allowed a cell viability of 100 % compared with controls, p > 0.05. Based on the results of cytotoxicity, we proceeded to evaluate the HIV infection inhibition with the aqueous extract at a concentration of 1 ppm and Azadirachtin at both concentrations. Inhibition percentages greater than 50 % were obtained and showed to be significant compared to controls p < 0.001 This work evidenced the HIV infection inhibition by aqueous neem leaf extracts and Azadirachtin; even though the antiretroviral mechanism is not completely understood. Furthermore, more studies need to be conducted in order to identify the active compound responsible for the anti- HIV activity described in the aqueous extract.

Recent Research on Akasha Nimba (Millingtonia hortensis)

• Chaiyasit, Sittiwet. (2009). Antimicrobial Activities of Millingtonia hortensis Linn. Flowers Essential Oil. Journal of Pharmacology and Toxicology. 4. 41- 44. 10. 3923/ jpt. 2009. 41. 44. Millingtonia hortensis Linn. flowers have been extracted for essential oil using vapor distillation, with 0.5- 2 % yield. The essential oil of M. hortensis Linn. was tested against various species of bacteria. The agar diffusion susceptibility test showed an inhibitory effect on 6 out of 10 tested strains. The growth of 4 gram-positive bacteria (S. aureus ATCC 25923, S. epidermidis ATCC 12228, B. subtilis ATCC 6633, and L. plantarum ATCC 14917) and 2 gram-negative bacteria (E. coli ATCC 25922 and P. vulgaris ATCC 13315) was inhibited by M. hortensis Linn. flower essential oil. The MICs (minimal inhibitory concentration) of M. hortensis Linn. flower essential oil are 0.5- 2 and 1- 4 ml L- 1, respectively. In this study, M. hortensis Linn. flower essential oil showed broad-spectrum antimicrobial activity at low concentration.
• Jetty, A. & Iyengar, D. S. (2000). Antimicrobial Activity of Millingtonia Hortensis Leaf Extract. Pharmaceutical Biology. 38. 157- 160. 10. 1076/ 1388-0209- 28200004- 293821- 1FT157. Polar extracts of the leaves of Millingtonia hortensis showed good antimicrobial activity. Twenty different bacterial strains and two yeast cultures were used. The aqueous alcohol extract showed good activity against all microbes tested, particularly Escherichia coli and Salmonella typhimurium, both Gram-negative bacteria, with MIC values of 6.25 µg/ml. The activity is compared with known antibiotics such as gentamycin and nystatin.
• Arumugam, Janaki & Muthu, Babu. (2017). Phytochemical screening of antioxidant and antibacterial activities of Millingtonia hortensis (L). International Journal of Current Pharmaceutical Review and Research. Herbal medicines are in great demand in both developed and developing countries because of their enhanced bio-efficacy and minimal side effects. In the present investigation, the leaf extracts of the plant Bauhinia tomentosa were evaluated for their cytotoxicity and antioxidant properties. The phytocompounds responsible for the bioactivity were analysed qualitatively and quantitatively. Phytochemical screening of the leaf extracts showed the presence of steroids, alkaloids, saponins, coumarins, flavonoids, phenols, and amino acids. GC-MS analysis revealed the presence of nine major phytocompounds in the methanol leaf extract. Antioxidant efficacy of the leaf extracts was analysed in vitro by DPPH and superoxide radical assays, and their cytotoxicity was evaluated on the MCF-7 cell lines. The leaf extracts were non-toxic, and their potent antioxidant activity and anticancer activity may be due to the presence of phytocompounds such as flavonoids and phenols.
• Puri, S, & Sharma, P.D. (2007). Antifungal activity of Millingtonia hortensis. Indian Journal of Pharmaceutical Sciences. 69. 10.4103/0250-474X.36959. Antifungal activities of different extracts of Millingtonia hortensis were investigated against various fungal pathogens. Methanol extract was found to have stronger activity than fluconazole against yeast-like fungi: 4-fold against Candida krusei with a 4 µg/ml minimal inhibitory concentration and 2-fold (MIC- 2 µg/ml) against Sacharomyces cerevisiae, though it showed the same activity as fluconazole against Candida glabrata. Aqueous extract also exhibited 4- fold stronger activity against Candida krusei (MIC- 4 µg/ ml) and 4- fold (MIC: 2 µg/ ml) against Sacharomyces cerevisiae. Chloroform and ethyl acetate extracts showed lower activities against all fungal pathogens except for Candida krusei, compared with the standard. Against the filamentous fungus, Trichosporon cutaneum, all extracts showed less activity than the standard.
• Babitha, S & Banji, David, & Banji, Otilia. (2012). Antioxidant and hepatoprotective effects of flower extract of Millingtonia hortensis Linn. on carbon tetrachloride-induced hepatotoxicity. Journal of pharmacy & bioallied sciences. 4. 307- 12. 10. 4103/ 0975- 7406. 103258. Millingtonia hortensis Linn is an abundant source of flavonoids, which might be beneficial in protecting liver tissue from injury. The hepatoprotective and antioxidant potential of ethanolic extract of M. hortensis on carbon tetrachloride (CCl (4)) induced hepatotoxicity, and the possible mechanism involved therein was investigated in rats.
• Preliminary phytochemical studies were carried out to determine the total phenol and flavonoid contents. 30 adult Wistar rats were allocated into 5 groups. Control group received vehicle, group-2 received CCl (4) alone (1 ml/ kg body weight, intraperitonially), groups 3 – 5 received the ethanolic flower extract in 2 dose levels (200 and 400 mg/ kg) and Curcumin (100 mg/kg) as a standard for 8 days orally, followed by CCl (4) as a single dose on the 8 (th) day. 48 hours later, blood was withdrawn, serum was subjected to biochemical assessments, and liver homogenate was examined for lipid peroxides, glutathione, superoxide dismutase, catalase, and total protein levels. Furthermore, hepatic tissues were subjected to histopathological studies. CCl (4) treatment produced a profound increase in the levels of malondialdehyde, hepatic marker enzymes, and bilirubin content compared with the control (P < 0.05). Pre-treatment with the flower extract of M. hortensis significantly enhanced the levels of endogenous antioxidants and reduced the levels of hepatic marker enzymes in relation to the CCl (4) treated group (P < 0.05). Balloning degeneration and fatty changes in hepatocytes were prevented by pre-treatment with the flower extract. The antioxidant nature of the flower extract of M. hortensis could be responsible for averting damage to the liver.
• Kumar, M. & Ravichandran, N. (2011). Cytotoxicity and antimicrobial studies of the stem bark of Millingtonia hortensis Linn. International Journal of Pharmacy and Pharmaceutical Sciences. 3. 247- 249. The present study aims to explore the cytotoxic potential and antimicrobial capacity of the aqueous extract of the stem bark of Millingtonia hortensis Linn. The aqueous extract of the stem bark of Millingtonia hortensis Linn is tested against the Human cervical cancer cell line by MTT Assay. The antimicrobial capacity of the extract is evaluated by means of the agar diffusion method using 12 strains of microbes. The extract is found to be a poor cytotoxic and antibacterial agent; however, it is found to be an effective antifungal agent. The isolation of phytoconstituent from the stem bark of Millingtonia hortensis Linn may lead to a novel antifungal agent rather than an antibacterial or cytotoxic agent.
• Tansuwanwong, Siwapong & Yamamoto, Hiroyuki & Imai, Kohzoh & Vinitketkumnuen, Usanee. (2007). ANTIPROLIFERATIVE EFFECT ON COLON CANCER CELL LINES BY AQUEOUS EXTRACT FROM THE BARK OF MILLINGTONIA HORTENSIS. 46. Millingtonia hortensis is a traditional medicinal plant widely used in Southeast Asia. This study focused on the antiproliferative effect when comparing aqueous extract and ethanol extract from the bark of M. hortensis using human colon cancer cell lines, DLD-1, HCT 15, SW48, and SW480. In in vitro experiments, the MTT method was used to determine the antiproliferative effect of M. hortensis extract on colon cancer cells. An aqueous extract of M. hortensis significantly reduced the proliferation rate of colon cancer cell lines in a dose-dependent manner. However, no antiproliferative effect was observed by the ethanol extract of M. hortensis. This finding indicated that only aqueous extract from the bark of M. hortensis can inhibit colon cancer cell proliferation, and its mechanism is now under investigation. Chiang Mai Medical Journal 2007; 46 (2): 61- 66.
• Jumnongprakhon, Pichaya & Pinkaew, Decha & Phuneerub, Pravaree. (2021). The antiaging property of the aqueous extract of Millingtonia hortensis flowers in aging neurons. Journal of Advanced Pharmaceutical Technology & Research. 12. 14. 10. 4103/ japtr. JAPTR_ 187_ 20. Cellular senescence is the key mediator of cellular dysfunction before undergoing degenerative disease, such as Alzheimer’s disease. The aging process was mainly mediated by the overactivation of senescence-associated β-galactosidase (SA-β-gal) enzyme, which led to several negative responses, including intracellular reactive oxygen species (ROS) production, cellular senescence regulation, and death, which contributed to encouraging synaptic loss. Thus, in the recent work, we evaluated the in vitro effects of aqueous extract of Millingtonia hortensis L. (MH) from the flowers in hydrogen peroxide (H2O2)- induced senescence in SK-N-SH cells. Herein, we demonstrated that MH significantly increased cell viability and decreased both of apoptotic cells and ROS production in a dose-dependent manner comparing to aging group (P < 0.01) using 3- (4, 5- dimethylthiazol- 2- yl)- 2, 5-diphenyltetrazolium bromide, flow cytometry, and ROS assay. Furthermore, the number of SA-β-gal-positive cells was also reduced in MH treatment (P < 0.01), together with the promotion of Sirt-1 protein. Importantly, MH also promoted synaptic plasticity by decreasing acetylcholinesterase activity and increasing synaptophysin expression in aging neurons compared to the aging group (P < 0.01). Hispidulin (the active ingredient in MH) was also revealed to have similar effects to MH. Therefore, we suggested that MH might be beneficial for neurodegenerative diseases caused by aging.
• Kumar, M. & Kavimani, S. (2014). Anthelmintic activity of Millingtonia hortensis Linn. F. stem bark on Indian adult earthworm. Der Pharmacia Lettre. 6. 89- 91. Millingtonia hortensis Linn. F (Bigoniaceae) is a traditional medicinal plant widely used in Southeast Asia for its fragrant flowers. It is commonly known as the Cork tree, Akash neem, and Neem chameli. The present study aims at exploring the anthelmintic potential of aqueous extract of the stem bark of Millingtonia hortensis Linn. F. An Anthelmintic study was carried out using the Indian earthworm, Pheretima posthuma. Anti-helmintic activity of Millingtonia hortensis aqueous extract was evaluated at 10 mg/ ml, 20 mg/ ml, 30 mg/ ml, 40 mg/ ml, and 50 mg/ ml, using piperazine citrate and piperazine hexahydrate as standards. The time taken for paralysis and death was calculated for all the groups. Millingtonia hortensis aqueous extract exhibited less anthelmintic activity as compared to the standard drugs. The study concludes that Millingtonia hortensis aqueous extract is not a suitable drug of choice for anthelmintic properties.
• Tansuwanwong, Siwapong & Hiroyuki, Yamamoto & Imai, Kohzoh & Vinitketkumnuen, Usanee. (2006). Induction of apoptosis in RKO colon cancer cell line by an aqueous extract of Millingtonia hortensis. Asian Pacific journal of cancer prevention: APJCP. 7. 641- 4. Millingtonia hortensis is an important medicinal plant in Southeast Asia, used for the treatment of asthma, sinusitis, and as a cholagogue and tonic. This study aimed to compare the effects of aqueous and ethanol extracts of Millingtonia hortensis on the induction of apoptosis in an RKO human colon cancer cell line. The viability of RKO cells was assessed by MTT reduction assay. The aqueous extract, but not the ethanol extract of M. hortensis, inhibited cell growth and proliferation in a dose- and time-dependent manner. Apoptotic cells were determined by flow cytometry and DNA fragmentation assay. Apoptotic cell numbers increased in a dose-dependent manner after treatment with aqueous extract. DNA ladders were clearly observed in RKO cells treated with 200, 300, and 400 g/ ml of the aqueous extract of M. hortensis for 48 h. These results indicate that the aqueous extract of M. hortensis inhibited cell proliferation in an RKO colon cancer cell line via the apoptosis pathway.