Shri Hari Mishra

Cytotoxicity evaluation and hepatoprotective potential of bioassay guided fractions from Feronia limmonia Linn leaf

OBJECTIVE To evaluate the cytotoxicity and hepatoprotective potentials of extracts, fractions or isolated compound from the leaves of Feronia limonia (F. limonia). METHODS Qualitative phytochemical analysis of extracts, fractions or compound was performed by means of thin layer chromatography and spectroscopic assays. The % purity of compound was measured by analytical HPLC. Extracts, fractions or compound have been individually evaluated for their cytotoxicity effects (10, 20, 100, 250, 500, 750 and 1 000 µg/mL). Based on the inhibitory concentration (IC50) obtained from the cell viability assay, graded concentrations of extracts, fractions or isolated compound were assessed (10, 20, 50, 100, 200 µg/mL) for its hepatoprotective potential against CCl4-induced hepatotoxicity by monitoring activity levels of serum glutamatic pyruvatic transaminase (SGPT) and serum glutamic oxaloacetic transaminase (SGOT). RESULTS Results indicated that the methanol extract of F. limonia was non-toxic and hepatoprotective in nature as compared with the petroleum ether extract. The acetone fraction of methanolic extract also showed similar properties but the subsequent two fractions were cytotoxic. However, the pure compound isolated from the penultimate fraction of methanolic extract was non-toxic and hepatoprotective in nature. Biochemical investigations (SGOT, SGPT) further corroborated these cytological observations. CONCLUSIONS It can be concluded from this study that F. limonia methanol extract, some fractions and pure isolated compound herein exhibit hepatoprotective activity. However, cytotoxicity recorded in the penultimate fraction and investigation of structural details of pure compound warrants further study.

A novel triterpenoid isolated from the root bark of Ailanthus excelsa Roxb (tree of heaven), AECHL-1 as a potential anti-cancer agent.

Background We report here the isolation and characterization of a new compound Ailanthus excelsa chloroform extract-1 (AECHL-1) (C29H36O10; molecular weight 543.8) from the root bark of Ailanthus excelsa Roxb. The compound possesses anti-cancer activity against a variety of cancer cell lines of different origin. Principal Findings AECHL-1 treatment for 12 to 48 hr inhibited cell proliferation and induced death in B16F10, MDA-MB-231, MCF-7, and PC3 cells with minimum growth inhibition in normal HEK 293. The antitumor effect of AECHL-1 was comparable with that of the conventional antitumor drugs paclitaxel and cisplatin. AECHL-1-induced growth inhibition was associated with S/G2-M arrests in MDA-MB-231, MCF-7, and PC3 cells and a G1 arrest in B16F10 cells. We observed microtubule disruption in MCF-7 cells treated with AECHL-1 in vitro. Compared with control, subcutaneous injection of AECHL-1 to the sites of tumor of mouse melanoma B16F10 implanted in C57BL/6 mice and human breast cancer MCF-7 cells in athymic nude mice resulted in significant decrease in tumor volume. In B16F10 tumors, AECHL-1 at 50 µg/mouse/day dose for 15 days resulted in increased expression of tumor suppressor proteins P53/p21, reduction in the expression of the oncogene c-Myc, and downregulation of cyclin D1 and cdk4. Additionally, AECHL-1 treatment resulted in the phosphorylation of p53 at serine 15 in B16F10 tumors, which seems to exhibit p53-dependent growth inhibitory responses. Conclusions The present data demonstrate the activity of a triterpenoid AECHL-1 which possess a broad spectrum of activity against cancer cells. We propose here that AECHL-1 is a futuristic anti-cancer drug whose therapeutic potential needs to be widely explored for chemotherapy against cancer.

Hepatoprotective potential of Tecomella undulata stem bark is partially due to the presence of betulinic acid.

ETHNOPHARMACOLOGICAL RELEVANCE Tecomella undulata (TU;` Family Bignoniaceae) is used in Indian Ayurvedic system of medicine for treating various diseases including hepatic ailments. It is also incorporated in various marketed hepatoprotective polyherbal formulations. AIM The present study was aimed at evaluating possible hepatoprotective role of isolated compounds from TU stem bark (TSB) using in vitro and in vivo experimental models. METHODS In vitro cytotoxicity and hepatoprotective potential of various extract, fractions and isolated compounds from TU stem bark were evaluated using HepG2 cells. Rats were pre-treated with TU methanolic extract (TSB-7) or betulinic acid (MS-2) or silymarin for 7 days followed by a single dose of CCl(4) (0.5 ml/kg, i.p.). Plasma markers of hepatic damage, hepatic antioxidants and indices of lipid peroxidation along with microscopic evaluation of liver were assessed in control and treatment groups. RESULTS TSB-2 and MS-1 accounted for significant cell death whereas; TSB-1, TBS-7, TSB-9, TSB-10 and, MS-2 did not register significant cytotoxicity. Further, non-cytotoxic components exhibited ascending grade of hepatoprotection in vitro (TSB-10<TSB-1<TSB-7<TSB-9<MS-2). Pre-treatment of TSB-7 or MS-2 to CCl(4) treated rats prevented hepatocyte damage as evidenced by biochemical and histopathological observations. CONCLUSION It can be concluded that, hepatoprotective potential of Tecomella undulata stem bark is partially due to the presence of betulinic acid.

Antioxidant markers based TLC-DPPH differentiation on four commercialized botanical sources of Shankhpushpi (A Medhya Rasayana): A preliminary assessment

Shankhpushpi is a cognition boosting traditional ayurvedic brain supplement. Convolvulus pluricaulis (Convolvulaceae), Evolvulus alsinoides (Convolvulaceae), Clitoria ternatea (Papilionaceae), and Canscora decussata (Gentianaceae) are botanical claimants of Shankhpushpi. This investigation is to focus the identification of the compound based on biological marker differentiation of four botanical claimants of Shankhpushpi for their antioxidant evaluation on thin layer chromatography (TLC) by 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. A rapid TLC-DPPH method was developed to identify and differentiate four botanical claimants of Shankhpushpi in terms of presence of β-carotene, rutin, scopoletin, chlorogenic acid, and mangiferin. C. pluricaulis shows presence of scopoletin; E. alsinoides shows presence of β-carotene, scopoletin, and chlorogenic acid; C. ternatea shows presence of β-carotene, scopoletin, and rutin; and C. decussata shows presence of β-carotene, scopoletin, and mangiferin. The order, they followed, based on their antioxidant potential is β-carotene < mangiferin < rutin < scopoletin < chlorogenic acid. Antioxidants are attributed for their beneficial role in age-related cognition decline. The proposed method provides an edge in terms of identification and quantification of antioxidant constituents in a multi-component system. This method may also provide application for identification of correct plant sources used in the name of Shankhpushpi in marketed ayurvedic formulation, food supplement, and extracts.

Traditional uses, phytochemistry and pharmacology of Tecomella undulata– A review

Abstract The aim of the present review is to present comprehensive information of the traditional uses, phytochemistry and pharmacology of Tecomella undulata (Family, Bignoniaceae) and to discuss future scope of research. Tecomella undulata (TU) is commonly known as desert teak (ver. Rohiro) and is traditionally for treating liver and spleen diseases, tumours, conjunctivitis, hepatosplenomegaly, syphilis, gonorrhea, hepatitis, as a blood purifier and in wound healing. Compounds such as naphthaquinone derivative, iridoid glucoside, phytosterol, fatty alcohol, flavonols, flavonoid glucoside and triterpenoids have been reported from TU. Anti HIV, anti bacterial, anti microbial, immune modulator, analgesic and hepatoprotective activities have been reported from its various aerial parts. In the present review, attempts have been made to compile research reports on TU, to assess current research trends with possible future avenues of research.

Hepatoprotective activity of Feronia limonia root

Objectives  The aim of this study was to evaluate the hepatoprotective potential of a methanolic extract and of marmesin isolated from the root bark of Feronia limonia.

Development and Validation of HPTLC Method for Quantitative Estimation of Oleanolic acid as Marker in Total Methanolic extract of Fruits of Randia dumetorum Lamk

The objective of the present investigation was to develop a validated HPTLC method for the determination of oleanolic acid as marker in the Methanolic extract of fruits of Randia dumetorum Lamk. Analysis of oleanolic acid was performed on TLC aluminium plates pre-coated with silica gel 60F-254 as the stationary phase. The mobile phase consists of Toluene: Ethyl acetate: Glacial acetic acid (7:3:0.1 v/v/v). Linear ascending development was carried out in twin trough glass chamber. The plate was sprayed with 10% sulphuric acid, heated at 110°C and immediately scanned at 540nm using Camag TLC scanner III. The system was found to give compact spots for oleanolic acid (R f value of 0.58 ± 0.01). The linear regression analysis data for the calibration plots showed good linear relationship with r 2 = 0.9922 ± 0.0002 in the concentration range 50-500ng per spot. The mean value (± S.D) of slope and intercept were 5.989 ± 0.0491 and 211.547 ± 4.5092 respectively. According to ICH guidelines the method was validated for precision, recovery, robustness and ruggedness. The limits of detection and quantification were 10 ng/spot and 30 ng/spot respectively. The oleanolic acid content of methanolic extracts was 3.45%. Recovery values from 99.38 - 100.79 % showed excellent reliability and reproducibility of the method. Statistical analysis of the data showed that the method is reproducible and selective. Since the proposed mobile phase effectively resolves oleanolic acid, the developed HPTLC method can be applied for identification and quantification of oleanolic acid in herbal extracts and formulations.

Pharmacognostic investigations on the leaves of Heterophragma quadriloculare K. Schum.

Objective To generate pharmacognostic parameters of leaves of Heterophragma quadriloculare K. Schum since data on pharmacognostic and phytochemical characterization which provide means to authentic raw material or drug for medicinally important formulation are not available.

Antidiabetic and antiplatelet aggregation study of various methanol fractions of Nymphaea stellata Willd. leaves

Summary Introduction: Nymphaea stellata Willd. (Nymphaeaceae) is traditionally used for the treatment of diabetes. Alcohol extract of N. stellata leaves has been reported for hypoglycaemic activity. Objective: The aim of this study was to further investigate the different methanol fractions of N. stellata leaves for anti-diabetic activity and anti-platelet aggregation activity. Methods: Methanol extract was fractioned in to unsaponified petroleum ether fraction of methanol extract (UPFME), chloroform fraction of methanol extract (CFME) and residual fraction of methanol extract (RFME). All fractions were evaluated for in vivo anti-diabetic activity (STZ-NAD-induced rat model), in vitro anti-diabetic activity (PTP1B inhibition study) and anti-platelet aggregation activity. Results: UPFME showed significant changes in all studied parameters, compared to the diabetic control. UPFME also showed an IC50 value of 19.30±1.1 mg/ml and 13.11±0.7 μg/ml in PTP1B inhibition study and anti-platelet aggregation study, respectively. Conclusion: The study indicates that UPFME of N. stellata leaves exhibit anti-diabetic and anti-platelet aggregation activity.