Md. Atiar Rahman

Antioxidative Potential of the Polyphenolics of Stephania japonica var. Discolor (Blume) Forman: A Chromatographic (High-Performance Liquid Chromatography) and Spectrophotometric Measure

This study investigated the quantitative phytochemical contents, total phenolics, total flavonoids, total carotenoids, antioxidative capacity, tannin, proanthocyanidins, lycopene, chlorophyll a, and chlorophyll b of the Stephania japonica extract. Comprehensive antioxidative effects of the extract were also investigated. Quantitative assays were conducted through both spectrophotometric and high-performance liquid chromatographic methods. Antioxidative effects were measured through FeCl3 reducing power, metal chelating power, reducing power, 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) scavenging activity, N, N-dimethyl-1,4-diaminobenzene free radical scavenging activity, superoxide radical scavenging activity, and nitric oxide scavenging effect. The contents of total phenolics, total flavonoids, total carotenoids, total antioxidative capacity, tannin, proanthocyanidins, lycopene, chlorophyll a, and chlorophyll b were found to be 47.32 ± 0.75 mg tannic acid equivalent, 61.41 ± 1.58 mg catechin equivalent, 63.29 ± 2.21 mg, 22.85 ± 0.70 mg ascorbic acid equivalent, 76.17 ± 0.97 mg tannic acid equivalent, 94.96 ± 4.49 mg catechin equivalent, 22.19 ± 0.79 µg, 22.19 ± 0.79 µg, 7.52 ± 1.24 mg, and 10.43 ± 2.11 mg, respectively, in 1 g of ethanol extract. A high concentration of epicatechin, p-coumaric acid, and rutin hydrate and moderate concentration of caffeic acid and quercetin was detected in the extract. The IC50 value for ferric reducing power assay, metal chelating assay, reducing power assay, ABTS scavenging assay, N, N-dimethyl-1,4-diaminobenzene scavenging assay, superoxide scavenging assay and nitric oxide (NO) assay were 465.06 ± 7.32 µmol ascorbic acid/g, 1656.52, 270.55, 457.27, 632.74, 217.5, and 464.00 µg/mL, respectively. No beta carotene was detected in the extract. The extract was demonstrated to be a very potential source of antioxidative metabolites.

Analgesic and anti-inflammatory effects of Crinum asiaticum leaf alcoholic extract in animal models

This study investigated the analgesic and anti-inflammatory effects of Crinum asiaticum (Amaryllidaceae) leaf ethanolic extract. Analgesic effect was investigated in acetic acid induced writhing model and formalin induced licking model in swiss albino mice. Anti-inflammatory effect was conducted in carrageenan-induced paw edema model of albino rat. Data were analyzed by one-way analysis of variance (ANOVA) followed by post hoc multiple comparison test. In analgesic study, C. asiaticum extract inhibited 42.34±3.20% of acetic acid induced pain at higher dose of 2.0 g/kg body weight. The effect was statistically significant (p<0.001) compared to the positive control, diclofenac sodium (10 mg/kg). The extract reduced the formalin induced pain 22.60±1.39% in early phase and 27.11±0.87% in late phase at the same dose of 2.0 g/kg and the reductions were significant (p<0.01) compared to the positive control morphine (0.5 mg/kg). In a time-dependent inhibition of carrageenan-induced paw edema model, the extract promoted the inhibitions of paw edema 51.60±2.50% at the 1st h and 40.80±0.52% at the 4th h of administration. These inhibitions were also significant (p<0.01) in comparison to those promoted by diclofenac sodium. No mortality was observed in acute toxicity test. The study concludes that C. asiaticum leaf extract has potential analgesic and anti-inflammatory effects to be recorded as plant-derived complementary medicine. Keywords : Crinum asiaticum , anti-inflammatory, analgesic, Carrageenan, formalin African Journal of Biotechnology Vol. 12(2)

Antioxidative Role of Hatikana (Leea macrophylla Roxb.) Partially Improves the Hepatic Damage Induced by CCl4 in Wistar Albino Rats

This research investigated the protective role of Leea macrophylla extract on CCl4-induced acute liver injury in rats. Different fractions of Leea macrophylla (Roxb.) crude extract were subjected to analysis for antioxidative effects. Rats were randomly divided into four groups as normal control, hepatic control, and reference control (silymarin) group and treatment group. Evaluations were made for the effects of the fractions on serum enzymes and biochemical parameters of CCl4-induced albino rat. Histopathological screening was also performed to evaluate the changes of liver tissue before and after treatment. Different fractions of Leea macrophylla showed very potent 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging effect, FeCl3 reducing effect, superoxide scavenging effect, and iron chelating effect. Carbon tetrachloride induction increased the level of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) and other biochemical parameters such as lipid profiles, total protein, and CK-MB. In contrast, treatment of Leea macrophylla reduced the serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) activities as well as biochemical parameters activities. L. macrophylla partially restored the lipid profiles, total protein, and CK-MB. Histopathology showed the treated liver towards restoration. Results evidenced that L. macrophylla can be prospective source of hepatic management in liver injury.

Antithrombotic Effects of Five Organic Extracts of Bangladeshi Plants In Vitro and Mechanisms in In Silico Models

This research was carried out to investigate the thrombolytic effects of the methanolic extracts of five Bangladeshi plants. Phytochemical metabolites of those plants have been identified to elucidate whether the plant-derived metabolites are linked with the thrombolytic effects. Potential computer aided models were adopted in this study to find out a structure-function correlation between the phytochemical constituents and thrombolytic effects using the secondary metabolites as ligands and tissue plasminogen activator (t-PA) as receptor for the best fit ligand-receptor interaction.

Evaluation of morning glory (Jacquemontia tamnifolia (L.) Griseb) leaves for antioxidant, antinociceptive, anticoagulant and cytotoxic activities

Abstract Background: The present study was planned to investigate the phytochemical, antioxidant, antinociceptive, anticoagulant and cytotoxic activities of the Jacquemontia tamnifolia (L.) Griseb leaf methanol extract (MExJT) in the laboratory using both in vitro and in vivo methods. Methods: Phytochemical values, namely, total phenolic and flavonoid contents, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging effect and FeCl3 reducing power effects, were studied by established methods. In vivo antinociceptive activity was performed by acidic acid-induced writhing test and formalin-induced pain test on Swiss albino mice at doses of 125, 250 and 500 mg/kg body weight. The clot lysis and brine shrimp lethality bioassay in vitro were used to evaluate the thrombolytic and cytotoxic activities of the plant extract, respectively. Results: Phytochemical screening illustrates the presence of tannins, saponins, flavonoids, gums and carbohydrates, steroids, alkaloids and reducing sugars in the extract. The results showed the total phenolic content (146.33 g gallic acid equivalents/100 g extract) and total flavonoid content (133.33 g quercetin/100 g). Significant (p<0.05) IC50 values compared to respective standards were recorded in DPPH radical scavenging (289.5 μg/mL) and FeCl3 reduction (245.2 μg/mL). The antinociceptive effect was evaluated in the acetic acid-induced writhing test and formalin-induced pain models in Swiss albino mice with doses of 125, 250 and 500 mg/kg body weight. Significant (p<0.05) inhibition (72.87±2.73%) of writhing response compared to diclofenac sodium was achieved by 500 mg/kg body weight. The extract also significantly inhibited the licking response in both the early phase (51.59±1.57%, p<0.05) and the late phase (64.82±1.87%, p<0.05) in the formalin-induced writhing test. MExJT also showed (38.10±1.79%) clot lytic activity in the thrombolytic test and cytotoxicity with an LC50 value of 31.70 μg/mL in the brine shrimp lethality bioassay. Conclusions: The plant is a potential source of antioxidants and might have one or more secondary metabolite(s) with central and peripheral analgesic activity. The results also demonstrate that MExJT has moderate thrombolytic and lower cytotoxic properties that may warrant further exploration.

Trypanosuppresive effects of ellagic acid and amelioration of the trypanosome-associated pathological features coupled with inhibitory effects on trypanosomal sialidase in vitro and in silico

BACKGROUND The search for novel antitrypanosomal agents had previously led to the isolation of ellagic acid as a bioactive antitrypanosomal compound using in vitro studies. However, it is not known whether this compound will elicit antitrypanosomal activity in in vivo condition which is usually the next step in the drug discovery process. PURPOSE Herein, we investigated the in vivo activity of ellagic acid against bloodstream form of Trypanosoma congolense and its ameliorative effects on trypanosome-induced anemia and organ damage as well as inhibitory effects on trypanosomal sialidase. METHODS Rats were infected with T. congolense and were treated with 100 and 200mg/kg body weight (BW) of ellagic acid for fourteen days. The levels of parasitemia, packed cell volume and biochemical parameters were measured. Subsequently, T. congolense sialidase was partially purified on DEAE cellulose column and the mode of inhibition of ellagic acid on the T. congolense sialidase determined. Molecular docking study was also conducted to determine the mode of interaction of the ellagic acid to the catalytic domain of T. rangeli sialidase. RESULTS At a dose of 100 and 200mg/kg (BW), ellagic acid demonstrated significant (P < 0.05) trypanosuppressive effect for most of the 24 days experimental period. Further, the ellagic acid significantly (P < 0.05) ameliorated the trypanosome-induced anemia, hepatic and renal damages as well as hepatomegaly, splenomegaly and renal hypertrophy. The trypanosome-associated free serum sialic acid upsurge alongside the accompanied membrane bound sialic acid reduction were also significantly (P < 0.05) prevented by the ellagic acid treatment. The T. congolense sialidase was purified to a fold of 6.6 with a yield of 83.8%. The enzyme had a KM and Vmax of 70.12mg/ml and 0.04µmol/min respectively, and was inhibited in a non-competitive pattern by ellagic acid with an inhibition binding constant of 1986.75μM. However, in molecular docking study, ellagic acid formed hydrogen bonding interaction with major residues R39, R318, and W124 at the active site of T. rangeli sialidase with a predicted binding free energy of -25.584kcal/mol. CONCLUSION We concluded that ellagic acid possesses trypanosuppressive effects and could ameliorate the trypanosome-induced pathological alterations.

Stigmasterol retards the proliferation and pathological features of Trypanosoma congolense infection in rats and inhibits trypanosomal sialidase in vitro and in silico

Stigmasterol has been reported to possess antitrypanosomal activity using in vitro model but information on the in vivo antitrypanosomal effects which is necessary in drug development process has not been evaluated. Hence, the present study investigates the in vivo effects of stigmasterol against T. congolense in addition to its inhibitory effects of trypanosomal sialidase. Stigmasterol, at 100mg/kg BW, did not significantly (p>0.05) reduce the progression of T. congolense infection in animals but a 200mg/kg BW stigmasterol treatment significantly (p<0.05) reduced the parasitemia, although, it did not completely eliminate the parasite from the bloodstream of infected animals. However, the stigmasterol treatments significantly (p<0.05) ameliorated the T. congolense induced anemia as well as hepatic and renal damages. Furthermore, the T. congolense-associated increase in free serum sialic acid with a corresponding decrease in membrane bound sialic acid were prevented, though insignificantly (p>0.05), by the 200mg/kg BW treatment. Subsequently, in vitro enzyme kinetic studies revealed that stigmasterol is an uncompetitive inhibitor of a partially purified bloodstream T. congolense sialidase with an inhibition binding constant of 356.59μM. Using molecular docking studies, stigmasterol formed a single hydrogen bonding interaction with a major residue (D63) at the catalytic domain of T. rangeli sialidase with a predicted binding free energy of -24.012kcal/mol. We concluded that stigmasterol could retard the proliferation and the major pathological features of T. congolense infection whilst the anemia amelioration was mediated via inhibition of sialidase.

In vitro antioxidative and cholinesterase inhibitory properties of Thunbergia grandiflora leaf extract

Abstract Among the pathologic hypotheses of Alzheimer’s disease (AD), cholinergic deficit and oxidative stress have been implicated as two major hallmarks. Therefore, inhibition of cholinesterase and oxidation are the two promising strategies in the development of a drug for AD. Thunbergia grandiflora leaf extract (TGEx) is used in this research to investigate its anticholinesterase and antioxidant potentials. Anticholinesterase activity was measured by modified Ellman method. Antioxidant potentials were evaluated by the assay of reducing power, radical scavenging and inhibition of lipid peroxidation. The Methanolic extract showed strong anticholinesterase effect. Additionally, the extract exhibited pronounced reducing capacity, radical scavenging ability, and lipid peroxidation inhibitory effect. The IC50 values of the extract for DPPH and hydroxyl free radical scavenging and lipid peroxidation were 10.50 ± 0.68, 24.98 ± 1.39 and 21.84 ± 0.91 μg/ml, respectively. Phytochemical screening of the extract revealed the presence of significant amount of total phenolics and flavonoids. The tested sample reflects potential antioxidative and anticholinesterase inhibitory effect which may warrant its effectiveness in the treatment of AD.

Antinociceptive and Anxiolytic and Sedative Effects of Methanol Extract of Anisomeles indica: An Experimental Assessment in Mice and Computer Aided Models

Anisomeles indica (L.) kuntze is widely used in folk medicine against various disorders including allergy, sores, inflammation, and fever. This research investigated the antinociceptive, anxiolytic and sedative effects of A. indica methanol extract. The antinociceptive activity was assessed with the acetic acid-induced writhing test and formalin-induced flicking test while sedative effects with open field and hole cross tests and anxiolytic effects with elevated plus maze (EPM) and thiopental-induced sleeping time tests were assayed. Computer aided (pass prediction, docking) analyses were undertaken to find out the best-fit phytoconstituent of total 14 isolated compounds of this plant for aforesaid effects. Acetic acid treated mice taking different concentrations of extract (50, 100, and 200 mg/kg, intraperitoneal) displayed reduced the writhing number. In the formalin-induced test, extract minimized the paw licking time of mice during the first phase and the second phase significantly. The open field and hole-cross tests were noticed with a dose-dependent reduction of locomotor activity. The EPM test demonstrated an increase of time spent percentage in open arms. Methanol extract potentiated the effect of thiopental-induced hypnosis in lesser extent comparing with Diazepam. The results may account for the use of A. indica as an alternative treatment of antinociception and neuropharmacological abnormalities with further intensive studies. The compound, 3,4-dihydroxybenzoic acid was found to be most effective in computer aided models.

Leea macrophylla Roxb. leaf extract potentially helps normalize islet of β-cells damaged in STZ-induced albino rats

Abstract This research aims to investigate the protective effects Leea macrophylla Roxb polyphenols on streptozotocin‐induced diabetic rats. Polyphenolic assays were undertaken through established methods. To conduct animal intervention study, forty Wistar albino male rats (average body weight 188.42 ± 7.13 g) of different groups were diabetized by streptozotocin (60 mg/kg) only in the animals of diabetic control (DC) and L. macrophylla extract (LM) groups. At the end of 4 weeks of intervention, serum was analyzed for insulin, liver and cardiac enzymes, lipid profiles, uric acid, and creatinine using ELISA method. In vitro α‐amylase inhibition of LM was evaluated and compared with reference drug acarbose. Pancreatic tissues were undertaken for histopathological screening. Food and fluid intake, weekly blood glucose level, liver glycogen, aspartate transaminase (AST), creatinine kinase (CK‐MB), cholesterol, and lactate dehydrogenase (LDH) were significantly decreased, whereas oral glucose tolerance (OGTT) ability, serum insulin concentration, and pancreatic islets morphology were significantly improved in the LM300 treatment group compared to the DC group. Alpha‐amylase inhibition was not found to be very promising for guiding the α‐amylase inhibition pathway. Results suggest that L. macrophylla can exert a potential effort to restore pancreatic β‐cell damaged by streptozotocin induction.