Biochemical and histological insights of 1,4-polyisoprene isolated from Sphenocentrum jollyanum pierre (menispermaceae) stem in wound healing activity in streptozotocin-induced diabetic rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Sphenocentrum jollyanum whole stem extract is used traditionally in combination with its leaves to treat chronic wounds and also ameliorate conditions that exacerbate wounds such as diabetes mellitus. AIM OF THE STUDY: The study isolated the major wound healing bioactive compound from the non-polar fraction of S. jollyanum extract and evaluated the in vivo wound healing activity of a 0.10% w/w 1,4-polyisoprene-based ointment in streptozotocin-induced diabetic rats. MATERIALS AND METHODS: The major bioactive constituent of S. jollyanum was isolated using a wound healing activity-guided approach and characterized the compound using 1D and 2D-NMR spectroscopic techniques. The wound healing activity study adopted both excision (wound contraction) and incision (biochemical) models. RESULTS: In the excision model, the 1,4-polyisoprene caused 99% wound closure and restored the excised wound on day 12. On the 6th and 12th post-wounding days, 1,4-polyisoprene caused a significant (p < 0.001) elevation in the tensile strength (486 g) of the incision wound compared with the control (388 g). The biochemical (hexosamine and hydroxyproline) and antioxidant/inflammatory (ascorbic acid, superoxide dismutase, and glutathione peroxidase) parameters increased significantly while malondialdehyde was down-regulated in the wounds treated with 1,4-polyisoprene compared with control. The histological analysis of tissue sections taken from the edge and center of the wounds at 0-12 days post wounding revealed an increased tissue regeneration, accelerated collagen formation, and epidermal regeneration without edema or inflammation on the 12th day. CONCLUSION: The major wound healing constituent of S. jollyanum is 1,4-polyisoprene and the study has provided a new class of compounds for further optimization.

Bioassay guided fractionation, isolation and characterization of hepatotherapeutic 1, 3-di-ortho-galloyl quinic acid from the methanol extract of the leaves of Pterocarpus santalinoides.

ETHNOPHARMACOLOGICAL RELEVANCE: Leaf extracts of Pterocarpus santalinoides DC are traditionally used to ameliorate ageing-related ailments such as heart and liver diseases, and have been reported to be protective against toxic injuries to the liver. AIM OF THE STUDY: This study aimed to isolate and characterize the hepatoprotective/hepatotherapeutic principle in the methanol leaf extract of P. santalinoides. MATERIALS AND METHODS: Fresh leaves of P. santalinoides were dried under shade and ground into powder. The ground leaves (2 Kg) were extracted with 80% methanol by maceration. Fractionation was carried out using column and thin layer chromatography techniques. Bioassay of fractions and sub-fractions was done using carbon tetrachloride (CCl4)-induced hepatotoxicity model in albino rats. Phytochemical analysis was carried out on the active compound. Characterization and structural elucidation of the active compound using high performance liquid chromatography and nuclear magnetic resonance spectroscopy was done. RESULTS: Extraction yielded 260 g dry extract. Six fractions (F1, F2, F3, F4, F5 and F6) were obtained after column and thin layer chromatography, with F6 (Rf = 0.78; Yield = 2.13 g) being the most active hepatotherapeutic fraction that significantly (p < 0.05) lowered serum ALT activity and increased serum albumin levels in CCl4-induced hepatopathy in albino rats. Further separation of F6 yielded four sub-fractions (F61, F62, F63 and F64), of which F61 with an Rf of 0.85 and a yield of 30.0 mg was isolated as the active hepatotherapeutic compound. Stiasny and ferric chloride test of F61 showed the presence of tannins in the fraction. Characterization of F61 revealed 1, 3-di-ortho-galloyl quinic acid. CONCLUSION: The hepatoprotective/hepatotherapeutic principle in the methanol extract of the leaves of P. santalinoides was identified as 1, 3-di-ortho-galloyl quinic acid.

Potential antidiabetic and antioxidant activities of a heliangolide sesquiterpene lactone isolated from Helianthus annuus L. leaves.

Heliangolide is a naturally occurring sesquiterpene lactone and its derivatives are biologically active compounds present in most medicinal plants. This study evaluated the antioxidant and antidiabetic properties of a heliangolide sesquiterpene lactone isolated from Helianthus annuus L. leaves. The heliangolide sesquiterpene lactone was isolated through a combination of solvent-solvent partitioning, column chromatography, thin layer chromatography and high-performance liquid chromatography techniques. The antioxidant activity of the compound was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and nitric oxide radical scavenging assays while the antidiabetic effects were investigated in alloxan-induced diabetic rats. The heliangolide derivative at the concentration of 954.2 µmol L-1 showed 23.7 % DPPH and 26 % nitric oxide radical inhibitions compared with 96.6 and 50.9 %, resp., displayed by the controls (2,271.2 µmol L-1). It also reduced the fasting blood glucose (FBG) levels in a time-dependent manner. The highest activity was recorded within 6 h post-treatment at 0.2 mmol kg-1 bm. The heliangolide derivative exhibited significant (p < 0.05) antioxidant and antidiabetic properties and provides a basis for further development of constituents of Helianthus annuus leaves for the management of such diseases.

Potent Antitrypanosomal Activities of 3-Aminosteroids against African Trypanosomes: Investigation of Cellular Effects and of Cross-Resistance with Existing Drugs.

Treatment of animal African trypanosomiasis (AAT) requires urgent need for safe, potent and affordable drugs and this has necessitated this study. We investigated the trypanocidal activities and mode of action of selected 3-aminosteroids against Trypanosoma brucei brucei. The in vitro activity of selected compounds of this series against T. congolense (Savannah-type, IL3000), T. b. brucei (bloodstream trypomastigote, Lister strain 427 wild-type (427WT)) and various multi-drug resistant cell lines was assessed using a resazurin-based cell viability assay. Studies on mode of antitrypanosomal activity of some selected 3-aminosteroids against Tbb 427WT were also carried out. The tested compounds mostly showed moderate-to-low in vitro activities and low selectivity to mammalian cells. Interestingly, a certain aminosteroid, holarrhetine (10, IC50 = 0.045 ± 0.03 µM), was 2 times more potent against T. congolense than the standard veterinary drug, diminazene aceturate, and 10 times more potent than the control trypanocide, pentamidine, and displayed an excellent in vitro selectivity index of 2130 over L6 myoblasts. All multi-drug resistant strains of T. b. brucei tested were not significantly cross-resistant with the purified compounds. The growth pattern of Tbb 427WT on long and limited exposure time revealed gradual but irrecoverable growth arrest at ≥ IC50 concentrations of 3-aminosteroids. Trypanocidal action was not associated with membrane permeabilization of trypanosome cells but instead with mitochondrial membrane depolarization, reduced adenosine triphosphate (ATP) levels and G2/M cell cycle arrest which appear to be the result of mitochondrial accumulation of the aminosteroids. These findings provided insights for further development of this new and promising class of trypanocide against African trypanosomes.