In-vivo evaluation of immunomodulatory activity of crude extract and solvent fractions of Cyphostemma adenocaule (Steud. ex A.Rich).

Background: Various indigenous medicinal systems including Ethiopians used Cyphostemma adenocaule for managing tumors, helminthiasis, snake envenomation, rabis virus, splenomegaly, and other immunological disorders. However, no scientific study was conducted to validate these traditional medicinal claims of the plant. Therefore, this study aimed to evaluate the in-vivo immunomodulatory activity of the crude root extract and its solvent fractions. Methods: Carbon clearance rate and humoral antibody titer were determined for 100, 200, and 400 mg/kg of crude extract and solvent fractions among Swiss albino mice. Carbon ink and sheep red blood cells were used as antigens for carbon clearance assay and humoral antibody titer respectively. Results: Among all groups, an increase in both carbon clearance rate and the humoral antibody titer was observed with an increase in the dose of both crude extract and solvent fractions. Compared to the solvent fractions of comparable doses, the crude extract showed better activity. The crude extract at a dose of 400 mg/kg revealed the highest and statistically significant augmentation of carbon clearance rate (0.1100 ± 0.0124) and humoral antibody titer (96.00 ± 14.31) compared to the vehicle control group. Conclusion: From our study, it is concluded that crude extract and n-butanol fraction showed promising immunostimulant activity by enhancing carbon clearance rate and humoral antibody titer.

Chemical Composition, Antioxidant Potential, and Blood Glucose Lowering Effect of Aqueous Extract and Essential Oil of Thymus Serrulatus Hochst. Ex Benth.

Thymus serrulatus, an endemic plant of Ethiopia, is traditionally used to cure various diseases and as a food ingredient. In the Ethiopian folk medicine, the decoction is orally taken as a remedy to treat diabetes and high blood pressure. The purpose of the present study was to evaluate the antioxidant and antihyperglycemic effects of the aqueous extract and of the essential oil of Thymus serrulatus. The chemical composition of the aqueous extract was determined by LC-MS and the essential oil was characterized by GC-MS analysis. Radical scavenging assays, namely scavenging of 2,2-diphenyl-1-picrylhydrazyl (DPPH•), hydroxyl (•OH), and nitric oxide (•NO), were used as a first approach to screen the potential antioxidant abilities of the samples. Alpha-amylase and α-glucosidase inhibitory studies were also employed to evaluate the in vitro antihyperglycemic potential of the plant. The in vivo blood glucose lowering effect of the extracts was assessed using hypoglycemic activity and the oral glucose tolerance test in normal and in streptozotocin induced diabetic mice. When compared to the aqueous extract, the essential oil showed superior radical scavenging activity, particularly for •NO, as well as greater inhibitory potency against α-amylase and α-glucosidase (IC50 = 0.01 mg/ml and 0.11 mg/ml, respectively). Both tested samples showed a statistically significant antihyperglycemic effect. The aqueous extract at 600 mg/kg exerted maximum antihyperglycemic activity (44.14%), followed by the essential oil (30.82%). Body weight and glucose tolerance parameters were also improved by the samples both in normal and diabetic mice. The findings of this study support the hypothesis that aqueous extract and essential oil of T. serrulatus are promising therapeutic agents.

Antihyperglycemic Activity of TLC Isolates from the Leaves of Aloe megalacantha Baker in Streptozotocin-Induced Diabetic Mice.

BACKGROUND: Diabetes mellitus (DM) is a group of metabolic disorders that are characterized by hyperglycemia which results from defects in insulin release or its efficient use by the human body. Although significant progress has been made to manage DM and related complications, it remains a major global health problem. To this end, the search for new antidiabetic drugs from traditionally claimed medicinal plants is important. Aloe megalacantha Baker is an endemic plant used traditionally to treat diabetes in Ethiopia. This study aimed to investigate antidiabetic activity of isolates from the leaf of A. megalacantha Baker in streptozotocin-induced diabetic mice. METHODS: The exudate of A. megalacantha was collected by cutting the leaves and scraping the yellow sap and then dried at room temperature. The dried exudate was subjected to repeated thin layer chromatographic (TLC) separations using a mixture of solvent system to isolate the major compounds identified on the TLC plate. A single dose of streptozotocin (50 mg/kg) was injected intraperitoneally to overnight fasted mice to induce diabetes. Antidiabetic activity of TLC isolates was assessed by in vivo methods including oral glucose tolerance test, antihyperglycemic and hypoglycemic activity tests. RESULTS: Three major isolates were obtained from the TLC analysis of the exudate of A. megalacantha. Exudate and TLC isolates were found to be non-toxic up to a dose of 2000 mg/kg. The TLC isolates (Ia and Ib) significantly (p<0.05) reduced blood glucose levels and also increased body weight change of the diabetic mice compared with control groups. CONCLUSION: The present study demonstrated the ability of the exudate of A. megalacantha and its TLC isolates to significantly decrease blood glucose levels and increase body weights in mice, thus strengthening the claim of its traditional use in DM-related disorders.

Evaluation of the Effect of Hydroethanolic Root Extract and Solvent Fractions of Cyphostemma adenocaule (Steud. ex A. Rich) Descoings ex Wild & Drummond on Cell-Mediated Immune Response and Blood Cell Count in Mice.

INTRODUCTION: Cyphostemma adenocaule (Steud. ex A. Rich) Descoings ex wild & Drummond (Vitaceae) has been used in traditional medicine for the management of various immunological and hematological disorders in different areas of Ethiopia and the rest of the world. In Ethiopia, the plant is used for the management of enlarged spleen, rabies virus, helminthic infection, snake bite, and various types of tumors. OBJECTIVE: To evaluate the effect of hydroethanolic root extract and solvent fractions of Cyphostemma adenocaule on cell-mediated immunity (delayed-type hypersensitivity), organ index (spleen and liver), and blood cell count in Swiss albino mice. MATERIALS AND METHODS: Acute oral toxicity test was conducted using nulliparous and nonpregnant Swiss albino mice following OECD 425 limit test method. Delayed-type hypersensitivity model was used to evaluate the effect on cell-mediated immunity. The experimental animals were divided into twelve groups which were sensitized and challenged with sheep red blood cells on day 0 and day 7, respectively. Levamisole 50 mg/kg was used as stimulant control, whereas cyclophosphamide 30 mg/kg was used as suppressant control. Hydroethanolic root extract (100 mg/kg, 200 mg/kg, and 400 mg/kg), aqueous fraction (100 mg/kg, 200 mg/kg, and 400 mg/kg), and n-butanol fraction (100 mg/kg, 200 mg/kg, and 400 mg/kg) were administered for seven days. The paw volume was measured using a digital plethysmometer before challenge and 24 hours after challenge. Blood was collected, and organs (spleen and liver) were isolated from each challenged mouse to determine blood cell count and organ index, respectively. RESULTS: No mortality and noticeable behavioral changes were observed among all mice receiving hydroethanolic root extract and solvent fractions at a dose of 2000 mg/kg. Hydroethanolic root extract and solvent fractions of Cyphostemma adenocaule showed enhancement of delayed-type hypersensitivity, organ index, and blood cell count. Hydroethanolic root extract at a dose of 400 mg/kg showed the highest and statistically significant stimulation of delayed-type hypersensitivity (0.123 ± 0.010) and blood cell count compared to the vehicle. CONCLUSION: Hydroethanolic root extract and solvent fractions of Cyphostemma adenocaule showed a stimulatory effect on cell-mediated immunity and hematopoiesis.

Analgesic, Anti-inflammatory and In-vitro Hyaluronidase Inhibitory Properties of the Leaf Extract and Solvent Fractions of Otostegia Fruticosa (Forssk.) Schweinf. ex Penzig.

Otostegia fruticosa is traditionally used to treat tonsillitis, stomach ache, asthma, arthritis, and febrile illness in different parts of Ethiopia and other countries. In this experiment 70% ethanolic crude extract and fractions of the leaf of Otostegia fruticosa (Forssk.) Schweinf. ex Penzig were evaluated for their in-vivo anti-inflammatory and analgesic activities and in-vitro hyaluronidase inhibition properties at different concentrations. Tail immersion, acetic acid induced writhing and carrageenan-induced paw edema model were used to assess the in-vivo analgesic and anti-inflammatory activities, respectively. Swiss albino mice of either sex were randomly divided into five groups of six mice per group and for evaluation of the fractions randomly divided into six groups of six mice per group. The test groups were treated with hydroalcoholic extract of Otostegia fruticosa (O. fruticosa) at doses of 100, 200, and 400 mg/kg. The positive control groups received either pethidine 5 mg/kg or aspirin at 100 mg/kg or 150 mg/kg. The negative control groups were orally given sunflower oil. All the fractions were administered at the dose of 400 mg/kg. In all models, the higher dose (400 mg/kg) of the crude extract and chloroform fraction showed a significant central and peripheral analgesic and anti-inflammatory activities with comparable effects to standards used. The hyaluronidase inhibition assay result showed that the test samples displayed concentration-dependent inhibitory activities. These findings indicate that 70% ethanol extract and organic solvent fractions of O. fruticosa leaves have potential analgesic, anti-inflammatory, and enzyme inhibitory activities.

In vivo Hepatoprotective and in vitro Radical Scavenging Activities of Extracts of Rumex abyssinicus Jacq. Rhizome.

BACKGROUND: Liver diseases contribute a prominent global burden of mortality and morbidity. The current therapies of liver diseases have numerous limitations including severe adverse effects. This denotes that new more effective, safer, and cheaper drugs are required and medicinal plants used in traditional medicines often offer ideal opportunities. Accordingly, the present study aimed to evaluate the in vivo hepatoprotective and in vitro radical scavenging activities of dried rhizome extracts of Rumex abyssinicus (R. abyssinicus), which is traditionally claimed to provide hepatoprotection. MATERIALS AND METHODS: Hepatoprotective activity of extracts was evaluated using carbon tetrachloride (CCl4)-induced liver injury in mice. Pre- and post-treatment models were employed to test the effect of the extracts and silymarin (standard drug). Serum biochemical markers and liver histopathology were used as parameters to evaluate hepatoprotective activities whereas in vitro radical scavenging activity was tested by 2, 2-diphenyl-2-picrylhydrazyl hydrate (DPPH) assay. RESULTS AND CONCLUSION: Oral administration of CCl4 (1 ml/kg) significantly (P<0.001) raised the serum levels of liver enzyme markers compared to the normal control group. Pre-treatment with 125, 250, and 500 mg/kg of R. abyssinicus extract reduced the serum level of CCl4-induced rise in liver enzyme markers with the highest reduction observed at a dose of 500 mg/kg. Likewise, in the post-treatment model, the crude extract and butanol fraction at dose 500 mg/kg reduced levels of liver enzymes. Histopathological examinations revealed lesser liver damage of extract-treated mice compared to the toxic (CCl4-treated) controls. The in vitro radical scavenging activity of the different extracts showed concentration-dependent radical scavenging activity. Thus, the results of this study may justify the traditional use of the plant as a hepatoprotective agent. CONCLUSION: Results of serum biochemical markers and histopathological examinations of CCl4-induced mice models, in the present study, show the hepatoprotective potential of extracts from the rhizome of R. abyssinicus.

Antidiabetic Activity of Extracts of Terminalia brownii Fresen. Stem Bark in Mice.

BACKGROUND: Diabetes mellitus is a chronic metabolic disorder that imposes a huge health and economic burden on societies. Because the currently available medications have many drawbacks, it is important to search for alternative therapies. Medicinal plants used in traditional medicines are ideal candidates. Hence, this study was undertaken to investigate the antidiabetic activity of crude extract and solvent fractions from the stem bark of Terminalia brownii Fresen. (Combretaceae) in mice. MATERIALS AND METHODS: The in vitro α-amylase inhibition assay was performed using the chromogenic 3, 5-dinitrosalicylic acid (DNSA) method while the antihyperglycemic activity was assessed using three mouse models: streptozotocin-induced diabetic mice, normoglycemic mice, and oral glucose challenged mice. Experimental diabetes was induced by a single intraperitoneal injection of streptozotocin at a dose of 150 mg/kg and animals with fasting blood glucose level (BGL) >200 mg/dL were considered diabetic. Glibenclamide (5 mg/kg) was used as a standard drug. Fasting BGL and body weight were used to assess the antidiabetic activity. The result was analyzed using GraphPad Prism software version 8 and one-way ANOVA followed by Tukey's post hoc test with p<0.05 considered as statistically significant. RESULTS: The crude extract of T. brownii at all tested dose levels (250, 500 and 750 mg/kg) showed a significant BGL reduction in all the three animal models. Moreover, the ethyl acetate and aqueous fractions (at 500 mg/kg) significantly (p<0.01) reduced the BGL in the streptozotocin induced diabetic model. The crude extract and different solvent fractions also showed a dose-dependent in vitro α-amylase inhibitory activity and improvement of body weight. CONCLUSION: T. brownii crude extract and its solvent fractions showed a significant antihyperglycemic activity in STZ induced diabetic mice, hypoglycemic activity and improvement of oral glucose tolerance in normal mice.

Antimalarial Activity of Meriandra dianthera Leaf Extracts in Plasmodium berghei-Infected Mice.

OBJECTIVE: To evaluate the antimalarial effect of aqueous methanolic extract and solvent fractions of Meriandra dianthera leaves against Plasmodium berghei in mice model. METHOD: M. dianthera leaves were extracted with 80% methanol and dried. The dried crude extract was then defatted and further fractionated with chloroform, ethyl acetate, and butanol. Acute oral toxicity test was performed as per the Organization for Economic Cooperation and Development guideline 425. Peter's 4-day suppressive test was used to determine the in vivo antimalarial activity of the extract and fractions. RESULT: The crude leaf extract of Meriandra dianthera leaves against P < 0.001) chemosuppression compared to the negative control. Both the extract and fractions were able to prevent P. berghei induced body weight loss and body temperature reduction and also increased the survival time of the mice as compared to the negative control. The aqueous methanolic leaf extract of M. dianthera leaves were extracted with 80% methanol and dried. The dried crude extract was then defatted and further fractionated with chloroform, ethyl acetate, and butanol. Acute oral toxicity test was performed as per the Organization for Economic Cooperation and Development guideline 425. Peter's 4-day suppressive test was used to determine the. CONCLUSION: The extracts of M. dianthera leaves showed promising antimalarial activity, with no sign of toxicity and therefore may support its traditional use for the treatment of malaria.M. dianthera leaves were extracted with 80% methanol and dried. The dried crude extract was then defatted and further fractionated with chloroform, ethyl acetate, and butanol. Acute oral toxicity test was performed as per the Organization for Economic Cooperation and Development guideline 425. Peter's 4-day suppressive test was used to determine the.

Evaluation of in vitro α-amylase inhibitory activity and antidiabetic effect of Myrica salicifolia in streptozotocin-induced diabetic mice.

The study was aimed to evaluate in vitro antioxidant, α-amylase inhibitory and in vivo antidiabetic activities of Myrica salicifolia root extracts. The powdered roots of M. salicifolia were extracted with 80% methanol and then dried. The dried extract was further fractionated into chloroform, ethyl acetate, butanol and aqueous fractions. The phytochemical screening of the crude extract was performed using standard chemical identification tests. The antioxidant activity of the extracts was determined by in vitro method using 2,2-diphenyl-1-picrylhydrazyl (DPPH) as radical scavenging reagent. The in vitro α-amylase inhibitory activity was performed using the chromogenic3,5-dinitrosalicylic (DNSA) method. The antidiabetic activity of M. salicifolia root crude extract (200, 400 and 600 mg/kg) and fractions (400 mg/kg) were evaluated in normal, glucose loaded hyperglycemic and streptozotocin (STZ)-induced diabetic mice. The crude root extract of M. salicifolia showed strong DPPH radical scavenging activity (IC50 = 4.54µg/ml) which was comparable with the standard antioxidant, ascorbic acid. In α-amylase inhibitory activity, the crude extract and butanol fraction showed highest enzyme inhibition. In the antidiabetic activity, daily administration of the crude extract, aqueous and butanol fractions for fifteen days showed highest significant reduction in fasting blood glucose level (BGL) compared to diabetic control in STZ-induced diabetic mice model. The root extract and fractions of M. salicifolia exhibited significant antihyperglycemic, α-amylase inhibitory and antioxidant activity with no sign of toxicity. The antidiabetic effect of the plant could be due to the synergistic effect of various classes of constituents present in the root part of the plant.

In vivo hepatoprotective and In vitro radical scavenging activities of Cucumis ficifolius A. rich root extract.

ETHNOPHARMACOLOGICAL RELEVANCE: Cucumis ficifolius A. Rich is a perennial prostrate herb that stems up to 1 m long. Its root is widely used in traditional medicine to treat various diseases including liver diseases. Yet, scientific evidence is lacking to verify its ethno medicinal claims. AIM OF THE STUDY: The present study was conducted to assess the hepatoprotective and radical scavenging activity of 80% methanol crude extract and different fractions of Cucumis ficifolius root. MATERIALS AND METHODS: Radical scavenging activity was done applying the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay while hepatoprotective activity was assessed using pre- and post-treatment models of carbon tetrachloride (CCl4)-induced liver injury in Swiss albino mice of either sex weighing 25-30 g. A single oral dose of 2000 mg/kg was used for acute toxicity study, doses of 125, 250, and 500 mg/kg were used in the pre-treatment model, and 500 mg/kg of extract and chloroform fraction were used in the post-treatment model. Biochemical markers and histopathological parameters were used to measure hepatoprotective activities. RESULTS: C. ficifolius crude extract and its solvent fractions showed strong radical scavenging activity and the chloroform fraction had the highest activity. No sign of toxicity was shown in an acute toxicity test of the extract. Hepatoprotective activity evaluation on the crude extract by a pre-treatment model with 125, 250, and 500 mg/kg doses revealed a significant (p < 0.05) reduction of the serum level of CCl4-induced liver enzyme markers at the highest tested dose (500 mg/kg). The chloroform fraction that had highest radical scavenging activity and the crude extract, both at 500 mg/kg, were again evaluated in a post-treatment model and the results revealed that both the extract and the chloroform fraction demonstrated significant (p < 0.05) hepatoprotective activities which support the results of the pre-treatment model. CONCLUSION: The present study verified the hepatoprotective potentials of C. ficifolius extract and its chloroform fraction which might be, at least in part, through radical scavenging action.