Prakash R. Itankar

Antidiabetic potential of unripe Carissa carandas Linn. fruit extract.

ETHNOPHARMACOLOGICAL RELEVANCE Carissa carandas commonly known as Karanda have a long history of use in traditional system of medicine. It is used by tribal healers of Western Ghat region of Karnataka as hepatoprotective and antihyperglycemic. However, no scientific data is available to validate the folklore claim. The present study has been designed to evaluate its unripe fruit for the antidiabetic activity. AIM In the present study, methanol extract of unripe fruits and its fractions were studied for its antidiabetic potential. MATERIALS AND METHODS The methanol extract and its fractions were screened for antidiabetic activity in alloxan induced diabetic rats. The polyphenolic, flavonoid and flavanone contents of methanolic extract and its fractions were also determined and correlated with its antidiabetic activity. RESULTS The experimental data indicated that the methanol extract and its ethyl acetate soluble fraction has significantly lowered the elevated blood glucose levels by 48% (p<0.001) and 64.5% (p<0.001) respectively at dose level of 400mg/kg per oral after 24h as compared to diabetic control. In order to assess the role of polyphenolic components in the relevant activity, polyphenolic and flavonoid contents were determined. The polyphenolic and flavonoid content of methanol extract and its ethyl acetate soluble fraction were found to be 15.8 ± 1.2mg and 18.55 ± 0.34 mg (gallic acid equivalent/g extract) and flavonoid content 2.92 ± 0.03 mg and 1.534 ± 0.30 mg (rutin equivalent/g extract) respectively. CONCLUSION The increased antidiabetic potential of ethyl acetate fraction over methanol extract is due to its partial purification achieved by fractionation which resulted in increase in degree of polymerization and segregation of secondary metabolites.

Evaluation of antidiabetic antihyperlipidemic and pancreatic regeneration, potential of aerial parts of Clitoria ternatea

The aim of the present study was to investigate the pancreatic regeneration potential of of diferent fractions of the ethanol extract Clitoria ternateaL., Fabaceae. The antidiabetic and antihyperlipidemic potential was evaluated in streptozotocin-induced diabetic rats and correlated with its in vivoand in vitro antioxidant activity. The extract and its fractions were initially screened for acute and sub-chronic antidiabetic activity in the dose range of 100200 mg/kg. The most potent extract and fractions were further evaluated for pancreatic β-cells regeneration activity along with antioxidant and antihyperlipidemic activity. The polyphenolic, flavonoid and flavanone contents were assessed and correlated with its antidiabetic activity. The most significant pancreatic regeneration activity, antidiabetic and antihyperlipidemic activity and was shown by ethanol extract and butanol soluble fraction at a dose level of 200 mg/kg, while rutin was found to be least potent. In conclusion, pancreatic regeneration studies of ethanol extract treated rats show nesidioblastosis. It is also suggested that the factors causing regeneration are present within the pancreas. The newly generated islets may have formed from the ductal precursor cells and reduced oxidative stress helps in restoration of β-cell function.

Anti-psoriatic potential of Solanum xanthocarpum stem in Imiquimod-induced psoriatic mice model

ETHNOPHARMACOLOGICAL RELEVANCE The plant Solanum xanthocarpum Schrad. & Wendl. (Solanaceae) is one of the members of the dashamula (ten roots) in Ayurvedic system of medicine. The stem and fruits are used as an antipyretic, antiasthmatic and is prescribed in skin infections and for relief in burning sensation in the feet accompanied by vesicular eruptions. OBJECTIVE To scientifically validate the anti-psoriatic potential of Solanum xanthocarpum stem in Imiquimod-induced psoriatic mice model. MATERIALS AND METHODS Ethanolic stems extract of Solanum xanthocarpum (ESX) was first subjected to phytochemical screening and quantification of identified phytoconstituents, which was further standardized with the help of HPTLC using chlorogenic acid as a marker. The extract was then subjected to acute oral toxicity and skin irritability study for determining the safety profile of the extract. Imiquimod-induced psoriatic mouse model was then performed to check the efficacy of extract against psoriasis, where treatment was carried out for 15 days both topically (Gel at 2.5%, 5% and 10%) as well as orally (at 100, 200 and 400mg/kg p.o.) and their Psoriasis Area Severity Index (PASI) was calculated. The study also included determination of levels of TNF-α, IL-1β, IL-6 and IL-17 in the animal tissues, which further included biochemical evaluations such as total collagen, hexosamine, hyaluronic acid DNA, protein antioxidant profiles such as lipid peroxidation, nitric oxide, superoxide dismutase and catalase along with histopathological studies of the tissues. RESULT ESX showed the presence of mainly phenols, tannins, flavonoids, alkaloids and carbohydrates, while chlorogenic acid was reported to be 3.49% w/w. The Imiquimod-induced psoriatic mouse model, depicted a potent anti-psoriatic activity of the extract both topical (10%) and oral (200 and 400mg/kg p.o., as evident through PASI grading The effect was found to be more prominent in case of topically treated as compared to orally treated mice. The results also showed a significant inhibition in the expression of TNF-α, IL-1β, IL-6 and IL-17 in treated animal tissues and also showed significant restoration of the altered biochemical parameters along with reduced hyperkeratinisation as observed through histopathology. CONCLUSION The study scientifically justified the anti-psoriatic activity of the ESX, which may be attributed to inhibition in the expression of cytokines such as TNF-α, IL-1β, IL-6 and IL-17. Further, the observed antioxidant, antimicrobial and cellular proliferative activities may act as a contributing factor in treatment of psoriasis, which may be attributed to the presence of chlorogenic acid along with other phytochemicals in combination.

Involvement of NFκB in the antirheumatic potential of Chenopodium album L., aerial parts extracts.

ETHNOPHARMACOLOGICAL RELEVANCE Chenopodium album L. (C. album) is commonly known as Bathua in Hindi (Family: Chenopodiaceae). Traditionally, the plant is used as a laxative, diuretic, sedative and the infusion of the plant is used for the treatment of rheumatism. However, no scientific validation is available on the antirheumatic potential of the plant. In the present investigation, role of NF kappa B (NFκB) in the antiarthritic potential of extracts of aerial parts of Chenopodium album was explored and evaluated. METHODS The defatted aerial parts of Chenopodium album were successively extracted with ethylacetate, acetone, methanol and 50% methanol to study their antioxidant capacity followed by antiarthritic potential using Complete Freund׳s adjuvant (CFA) induced arthritis model in rats. The polyphenol, flavonoid and flavanone contents of different extracts were quantified and correlated with their antioxidant capacity, antiarthritic activity and NFκB inhibition potential. RESULTS The experimental data indicated that the acetone extract of Chenopodium album (ACCA) has shown significant reduction in rat paw edema (80.13%) at dose level of 200 mg/kg per oral in 21 days of this study. On 22nd day, hematological and biochemical parameters were estimated and it was observed that the altered hematological parameters (Hb, RBC, WBC and ESR), biochemical parameters (Serum creatinine, total proteins and acute phase proteins) and loss in body weight in the arthritic rats were significantly brought back to near normal level by the ACCA extract. ACCA extract significantly decreased the NFκB expression in paraventricular nucleus of hypothalamus and this effect is comparable with standard indomethacine in CFA treated rats. The polyphenolic and flavonoid content of different extracts were in the range of 14.56±0.21-42.00±0.2 mg (gallic acid equivalent/g extract) and 2.20±0.003-7.33±0.5 mg (rutin equivalent/g extract) respectively. CONCLUSION The antiarthritic activity possessed by ACCA extract can be correlated directly to its antioxidant potential, high flavonoidal content achieved by successive extraction and its capacity to inhibit the NFκB protein, as proven by immunohistochemistry study.

Antioxidant potential and its relationship with polyphenol content and degree of polymerization in Opuntia elatior Mill. fruits

Background: Opuntia elatior Mill. (Nagaphani) fruits are traditionally recommended as an expectorant, remedy for whooping cough, asthma, gonorrhea, ulcers, tumors, in the treatment of diarrhea and syphilis. Many of these diseases are allied with oxidative stress caused by free radicals. Thus, current research is directed towards finding naturally-occurring antioxidants of plant origin. Aim: To evaluate antioxidant potential of hydro-alcoholic extract of the O. elatior fruits (HAOE) and its fractions. Materials and Methods: Using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and nitric oxide radical scavenging assay, total polyphenolic, flavonoid (FA), flavanone (FO) contents and degree of polymerization in relation with its antioxidant activity were examined. Results: The experimental data indicated that the HAOE, ethyl acetate (EAOE) and butanol (BFOE) soluble fractions have shown significant antioxidant activity. The highest polyphenolic, FA, FO contents and degree of polymerization were found in EAOE. The scavenging potential was in the order of Ascorbic Acid > EAOE > BFOE > HAOE > BIOE, where ascorbic acid was used as a positive control. The increased antioxidant potential of EAOE and BFOE fractions over HAOE extract may be attributed to the purification achieved by fractionation of the extract which in turn resulted in an increase in the degree of polymerization and segregation of secondary metabolites. Conclusion: The fruit of O. elatior can be used as the best alternative for synthetic antioxidants.

Antidiabetic Potential and its Relationship with Polyphenol and Degree of Polymerization in Acalypha indica Linn Leaves

Acalypha indica commonly known as Kuppi belongs to family Euphorbiaceae. Traditionally the plant used as a laxative, anthelmintic, cathartic and the juice is used as a speedy emetic for children and expectorant. It is used by tribal areas of Maharashtra State as rheumatic arthritis and as antihyperglycemic. However, no scientific data is available to validate the ethnobotanical claim. Keeping the above information in view, the present study has been designed to evaluate its antidiabetic activity. In the present study, hydroalcoholic extract of leaves and its fractions were studied for its anti diabetic potential in alloxan induced diabetic rats. The poly phenolic, flavonoid and flavanone contents of hydroalcoholic extract and its fractions were also determined and correlated with its anti diabetic activity. The experimental data indicated that the hydroalcoholic extract and its chloroform soluble fraction has significantly lowered the elevated blood glucose levels by 36.82% and 61.52% respectively at dose level of 400 mg/kg per oral after 7 days as compared to diabetic control. The poly phenolic and flavonoid content of hydro alcoholic extract and its chloroform soluble fraction were found to be 7.9 ± 0.20 mg and 1.14 ± 0.21mg (gallic acid equivalent/ g extract) and flavonoid content 2.02 ± 0 .50 mg and 0.56 ± 0.003mg (rutin equivalent/g extract) respectively. The increased anti diabetic potential of chloroform fraction over hydro alcoholic extract is due to its partial purification achieved by fractionation which resulted in increase in degree of polymerization and segregation of secondary metabolites.

Extraction, isolation and identification of flavonoid from Chenopodium album aerial parts

Chenopodium album L., (C. album) (family: Chenopodiaceae) is an annual shrub widely grown in Asia, Africa, Europe and North America. It is commonly known as Bathua (in Hindi), pigweed, fat hen or lamb-quarters. The leaves of C. album are applied as a poultice to bug bites, sunstroke, rheumatic joints and as mild laxative. The flavonoids contained in C. album aerial parts were extracted, identified and characterized. Sequential soxhlet extraction was subjected to preliminary phytochemical screening and flavonoid quantification. The results showed that maximum yield of the flavonoid (7.335 mg/g) were obtained from acetone extract. This acetone extract was subjected to flash chromatography for isolation of flavonoid. Characterization of isolated flavonoid was done by UV, IR, 1H & 13C NMR and MS. On the basis of chemical and spectral analysis structure was elucidated as 2-(3, 4-dihydroxyphenyl)-3, 5, 7-trihydroxy-4H-chromen-4-one, a flavonoid.

Toxicological and pharmacological profiling of organically and non-organically cultivated Cymbopogon citratus

Background With the rapid depletion of forests, impairing the availability of raw drugs, Ayurveda, has reached a very critical phase. Consequently, cultivation of medicinal plants is essential to ensure their availability to the industry. In view of the above concept, organic farming of medicinal plants needs scientific validity. Objectives The present study includes the organic and non-organic cultivation of Cymbopogon citratus, followed by toxicological and pharmacological profiling of extracts. Materials and methods C. citratus was simultaneously cultivated organically (OCC) and non-organically (NCC). Toxicity profile of aqueous extracts was recorded on prokaryotes using bioluminescent bacteria, Vibrio harveyi and evaluated its type 2 anti-diabetic activity. Results OCC have shown the higher mean values of height, number of germplasms and root lengths compared to NCC. The higher level of toxicity was shown by NCC with decrease in bioluminescence with increasing concentration of extract. In acute type 2 anti-diabetic study, OCC showed prominent decrease in blood glucose at postprandial condition (6th h) (48.86% OCC-200). The order of sub-chronic anti-diabetic activity was observed as positive control > OCC-200 > NCC-200, while OCC at 200 mg/kg corrected the altered lipid profile and antioxidant status with significant increase in body weights of animals. Histopathological examination of pancreas showed the enlargement of pancreatic islets and formation of neo islets with degenerative changes in OCC treated animals. Conclusion The study confirms that organically grown C. citratus is better in terms of nourishment, biological activity and safety measures.

High performance thin layer chromatography fingerprinting, phytochemical and physico-chemical studies of anti-diabetic herbal extracts

Introduction: Herbal medicines have gained increasing popularity in the last few decades, and this global resurgence of herbal medicines increases their commercial value. However, this increasing demand has resulted in a decline in their quality, primarily due to a lack of adequate regulations pertaining to herbal medicines. Aim: To develop an optimized methodology for the standardization of herbal raw materials. Materials and Methods: The present study has been designed to examine each of the five herbal anti-diabetic drugs, Gymnema sylvester R. Br., Pterocarpus marsupium Roxburgh., Enicostema littorale Blume., Syzygium cumini (L.) Skeels. and Emblica officinalis Gaertn. The in-house extracts and marketed extracts were evaluated using physicochemical parameters, preliminary phytochemical screening, quantification of polyphenols (Folin–Ciocalteu colorimetric method) and high performance thin layer chromatography (HPTLC) fingerprint profiling with reference to marker compounds in plant extracts. Results: All the plants mainly contain polyphenolic compounds and are quantified in the range of 3.6–21.72% w/w. E. officinalis contain the highest and E. littorale contain the lowest content of polyphenol among plant extracts analyzed. HPTLC fingerprinting showed that the in-house extracts were of better quality than marketed extracts. Conclusion: The results obtained from the study could be utilized for setting limits for the reference phytoconstituents (biomarker) for the quality control and quality assurance of these anti-diabetic drugs.

OA01.47. Phytochemical study and evaluation of antileukemic activity of ripe fruit of Opuntia elatior Mill

Purpose: Opuntia elatior Mill. (Family: Cactacea) is a xerophytic perennial bushy shrub, 1-3 m tall. It is native of Caribbean, Central America and northern South America, but actually it is fully naturalized in many part of the world like Mediterranean, Australia, Africa and India. Ripe fruits of this plant contain red pigment i.e. betanin which is betacyanin pigment. It is recently documented that natural food colors such as betanin can inhibit the cell proliferation of a variety of human tumor cells. In view of its traditional use and chemical composition, Opuntia elatior Mill. fruit was selected for its in vitro antileukemic and antioxidant activity. Method: The fruits of Opuntia elatior after collection, authentication and drying was extracted with hydroalcoholic solvent using maceration method. The present study aimed to determine the preliminary phytochemical evaluation, quantitative estimation of Betanin pigment, total phenolic, flavonoid and flavonone content with spectrophotometric methods, evaluation of antioxidant and cytotoxicity study of the hydroalcholic extract of Opuntia elatior on K-562 cell line (Human chronic myelocytic leukemia) by MTT assay method. Result: The preliminary phytochemical evaluation showed the presence of carbohydrates, flavonoids, betanin pigment, tannins, sterols, proteins, pectin, citric acid and vitamin C. The total phenolic, flavonoid, flavonone and betanin content in extract were found to be 52.76 mg/g, 39.22 mg/g, 9.60 mg/g and 48.10 mg/100ml of extract respectively. Evaluation of antioxidant activity was carried out by DPPH free radical scavenging method. It showed significant activity compared with the standard. IC50 values for extract of O.elatior and Vitamin C was found to be 88.16 g/ml and 62.83 g/ml respectively. Conclusion: Its cytotoxic evalution showed a potent action against to K-562 (Human chronic myelocytic leukemia) cell line. It was found that hydroalcoholic extract of the fruits of Opuntia elatior Mill. have persuasive antioxidant activity and promising antileukemic activity.