Effect of phytoestrogen-rich fraction of Millettia aboensis on lipid profile, oxidative stress, and platelet count in ovariectomized rat model of menopause

Milletia aboensis has long-standing ethnopharmacological indications for the treatment of symptoms and diseases related to menopause. This investigation examined the effects of its phytoestrogen-rich fraction on some predictors of cardiovascular risk in an ovariectomized rat model of menopause. In vitro, antioxidant activity was determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) test. Ovariectomy was used to trigger menopause by surgically removing the ovaries under anesthesia. After recovery, the animals were treated orally with the phytoestrogen-rich fraction (PERF) of M. aboensis daily for 30 days. Blood samples were obtained for the estimation of serum lipid profile, antioxidant enzyme activities, lipid peroxidation, and platelet count. The PERF showed the strongest inhibition of DPPH radical with an IC50 value of 19.65 µg/ml. At 200 and 400 mg/kg, the PERF dose-dependently showed significant increase in high-density lipoprotein (HDL) (42.86 and 43.05 mg/dl) and a significant (p < 0.05) decrease in low-density lipoprotein (LDL) levels (3.78 and 1.24 mg/dl) compared with the ovariectomized (OVX) control (31.82 and 20.32 mg/dl, respectively). Being dose-dependently, the PERF increased catalase (CAT) and superoxide dismutase (SOD) enzyme activities as well as significantly reduced malondialdehyde (MDA) levels compared to OVX control. At 200 and 400 mg/kg, the PERF restored platelet count with values (316.54 and 343.3 103/IU, respectively) close to that of sham-operated control (365.17 103/IU). The ability of M. aboensis to reverse lipid abnormalities associated with ovariectomy coupled with its reduced platelet count, and antioxidant effects make it a potential therapeutic phytoestrogen remedy for the management of cardiovascular complications associated with estrogen deficiency

Antioxidant Activities of the Leaf Extract and Fractions of Dryopteris filix-mas (L.) Schott could be Attributed to The Abundance of Polyphenol Compounds

Dryopteris filix mas (D filix-mas) is wildly used in ethnomedicine for the management of rheumatoid arthritis, wounds and other diseases. We investigated the anti-oxidant activities of its leaf extract, and chromatographic fractions. The ethanol leaf extract was partitioned into four fractions; n-hexane, ethyl acetate, n-butanol and water. Ferric reducing anti-oxidant power (FRAP), 1, 1-diphenyl-2-picrylhydrazil (DPPH) and nitric oxide (NO) scavenging in vitro assays were carried out on the extract and fractions at 6.25, 12.5, 25, 50, 100, 200, 400 and 800 µg/mL. The most active fraction (ethyl acetate fraction) was further purified using chromatographic techniques to isolate its major compound whose structure was elucidated using ID nuclear magnetic resonance (NMR) and mass spectrometry. The ethyl acetate fraction produced the highest free radical scavenging activity among the other fractions. The fraction (VLC-E7) from which the bioactive compound, quercetin-3-O-αL-rhamnopyranoside, was isolated had the best FRAP and DPPH scavenging activities with EC50 and IC50 values of 88.81 ± 3.41 and 26.87 ± 0.24 respectively more than the ethyl acetate fraction. This study revealed that the polyphenol flavonoid, quercetin-3-O-αL-rhamnopyranoside could be responsible for antioxidant activity of ethno-medicinal property of D filix-mas leaf

In silico and in vitro investigations of antihelmintic activities of selected approved drugs

Objective: We investigated the binding affinities of some approved drugs to Ascaris suum Mitochondrial Rhodoquinol Fumarate Reductase (MRFR), an essential enzyme for ascaris survival, and the possibility of repurposing these drugs as antihelmintic agents using in silico molecular docking and in vitro paralysis and mortality times of fifteen selected front runners.Method: Two hundred approved drugs were selected from ZINC® database based on bioactivity scores while MRFR (PDB code, 3vra) was obtained from the Protein Data Bank (PDB). Both were prepared using AutoDock tools v.1.5.6 and Chimera v.1.9.The docking protocol was validated by computationally reproducing the binding of atpenin to MRFR. The selected approved drugs and the receptor were docked using AutoDockVina v. 4.0. The docking results were analyzed using PyMoL v. 1.4.1.The paralysis and mortality times of the identified frontrunners against Pheretima posthuma were determined in vitro and synergistic testings were done by the checkerboard method.Result: Fifteen drugs had binding free energies between -7.825 to -11.025 kcal/mol while four of these drugs (mefloquine, doxycycline, mepacrine and proguanil) emerged as major frontrunners by both in silico and in vitro assessments. The paralysis and mortality times of the four drugs were between 0.33-0.50 hr as against 1.80- 2.36 hr for albendazole. They were therefore predicted to have ability to affect MRFR in the same manner as atpenin hence, suggestive of potential antihelmintic activity.Conclusion: The antihelmintic potentials of mefloquine, doxycycline, proguanil and mepacrine have been demonstrated. In vivo investigation of these frontrunner drugs is strongly recommended.</p

Osteoprotective Effect of Phytoestrogen Rich Fraction of Ochna schweinfurthiana Stembark Extract on Ovariectomized Animal Model of Menopause

Osteoporosis is one of the major disease conditions associated with estrogen deficiency in postmenopausal women. This study evaluated the osteoprotective effect of phytoestrogen rich fraction (PERF) of O. schweinfurthiana back extract on ovariectomized animal model of menopause. The methanol plant extract was partitioned into n-hexane, ethyl acetate, butanol and water fraction using liquid-liquid fractionation. The phytoestrogens in the fractions were quantified using genistein as standard. Ovariectomy induced osteoporosis was achieved by surgical removal of the ovaries. Body weight changes, relative uterine weight, serum estradiol, bone turnover markers (Alkaline phosphatase - ALP and Acid Phosphatase – ACP), bone mineral content and density as well as lipid peroxidation assays were used to establish ability of ovariectomy to induce osteoporosis and dose response effect of the PERF. Water fraction showed the highest content of phytoestrogen which was more than 10 fold higher than the content of other fractions. Ovariectomy induced weight gain was significantly (P&lt;0.05) lowered by PERF compared to vehicle control. Significant (P&lt;0.05) increase in serum estradiol following treatment with PERF produced corresponding uterotrophic effect with median effective dose (ED50) of 197 mg/kg. osteoprotective effect of PERF was evident from the significant (P&lt;0.05) increases in both bone mineral content and density which corresponded with reduction in bone turnover markers. Ovariectomy induced oxidative stress was reduced by PERF with ED50 of 716 mg/kg. O. schweinfurthiana exhibited osteoprotective effect that may have been mediated by downstream signaling effects through estrogen receptor. Keywords: Osteoporosis; Menopause; Phytoestrogen; bone turnover DOI: 10.7176/JNSR/14-10-04 Publication date:July 31st 202

Combination Interactive Effects of Gongronema latifolium Leaves and Picralima nitida Seeds Extracts on Glucose Tolerance

This study evaluated the combination interactive effects of G. latifolium leaves and P. nitida seed extracts using a metabolic glucose tolerance test. The plant samples were extracted separately using cold maceration and their acute toxicities were determined. Dose-response glucose tolerant tests of both plants were done using a 2 g/kg glucose load monitored over 0 – 1h. A 41% effect isobologram was used to determine the needed dose combinations according to the principle of Loewe’s additivity model. The glucose tolerant tests of dose pairs of the combined extracts were evaluated and their combination indices were calculated to determine the nature of their interaction. The ED41 of G. latifolium (GL) and P. nitida (PN) were 180 mg/kg and 254 mg/kg respectively. The percentage reductions of GL:PN (50:160); GL:PN (100:90); and GL:PN (150:30) dose pairs were 48.23, 50.76 and 42.99 % respectively. Their combination index were calculated to be 0.91, 0.91 and 0.95 respectively - an indication of synergistic interaction. Findings from this study validate the combined use of G. latifolium leaves and P. nitida seeds in folkloric medicine. However, combining the extracts of G. latifolium: P. nitida in the dose ratios of 50:160, 100:90 and 150:30 mg/kg gave the best dose pairs with synergistic outcome

Evaluation of the in vivo antioxidant, toxicological and chromatographical profiling of leaf extract and fractions of Crescentia cujete Linn. (Bignoniaceae)

Oxidative stress has been reported to be involved in many diseases pathogenesis. This study evaluated the antioxidant potentials of Crescentia cujete leaf. The leaves were extracted in methanol by cold maceration and fractionated into n-hexane (NHF), ethyl acetate and butanol fractions. The in vitro antioxidant activity was evaluated using 2,2-diphenyl-1-picrylhydrazil (DPPH) and ferric reducing antioxidant power (FRAP) tests while Catalase (CAT), Superoxide dismutase (SOD) and Lipid peroxidation (LPO) assays were used to evaluate in vivo antioxidant potentials. The constituents of the extract and fractions where analyzed using high performance liquid chromatography coupled to diode array detector (HPLC-DAD). The median lethal dose (LD50) of CME and serum liver marker (SLM) enzymes were evaluated using standard protocols. The crude methanol extract (CME) and the fractions showed concentration dependent in vitro antioxidant activity (EC50 values of 15.22 to 569.22 μg/ml for DPPH and 54.69 to 581.40 μg/ml for FRAP) with ethyl acetate fraction (EAF) showing the highest activity. The CME at 400 mg/kg and EAF at 200 mg/kg showed more than 3-fold increase in CAT and SOD activities compared with the vehicle treated group. EAF (400 mg/kg) significantly (p&lt; 0.05) inhibited LPO. HPLC-DAD analysis of CME and EAF revealed the presence of phenolic compounds (chlorogenic acid, protocatechuic acid and several quercetin derivatives). The LD50 of CME was above 5000 mg/kg. The CME and EAF showed significant (p&lt; 0.05) reduction of SLM enzymes. The study validated the antioxidant activity of C. Cujete, which may be connected to high abundance of the detected phenolic compounds

Wound-healing Activity of the Aqueous Leaf Extract and Fractions of Ficus exasperata (Moraceae) and its Safety Evaluation on Albino Rats

Ficus exasperata have been reported to have wide applications in the treatment of many human diseases. However, its traditional use in the treatment of wounds has not been validated by any scientific study. Also, its safety in the management of chronic disease conditions requires attention. We evaluated the wound-healing activity of the aqueous extract and fractions of F. exasperata, as well as its safety after subchronic oral administration. Similar percentage of wound contraction was observed with 5% w/w extract ointment application and administration of cicatrin powder (standard) on the 4th day, while better contraction than the standard was recorded with higher concentrations of the extract ointment. Of all the fractions tested, significant (P<0.05) contraction was only noticed in chloroform fraction, though lower than that of the aqueous extract. The extract also showed concentration-dependent inhibition of all the tested microbial isolates. Extract administered up to 5000 mg/kg (single dose administration) did not cause any mortality after 24 h. Mortality was, however, recorded at 4000 mg/kg within the first 20 days of subchronic administration of the extract. Significant (P<0.05) increases in alanine aminotransaminase (ALT), aspartate aminotransaminase (AST), and in particular, alkaline phosphatase (ALP) were observed at different doses and time periods. Pathological and histological changes were noticed in the liver and kidney on the 91st day of the study with 4000 mg/kg of the extract. Except for the significant (P<0.05) reduction in WBC on the 91st day, no other significant (P<0.05) changes were observed in other hematological parameters. The aqueous extract demonstrated better wound-healing activity than its fractions; however, the extract may not be safe at higher doses for subchronic oral administration, as may be the case in the management of chronic disease conditions

In vitro and in vivo antioxidant potentials of Alchornea floribunda leaf extract, fractions and isolated bioactive compounds

Objective: Alchornea floribunda leaves are widely used in ethnomedicine for the management of immuno-inflammatory disorders. We investigated the in vivo and in vitro antioxidant activity of the leaf extract, fractions and isolated compounds of A. floribunda.Materials and Methods: The ethyl acetate fraction of the methanol leaf extract was subjected to several chromatographic separations to isolate compounds 1-4. The structures of the isolated compounds were elucidated by a combination of 1D and 2D NMR and mass spectrometry. Oxidative stress was induced with carbon tetrachloride (CCl4). Further analysis on the isolated phenolic compounds were done using 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP) and hydrogen peroxide scavenging activity tests.Results: The ethyl acetate fraction at 200 mg/kg produced significant (