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.

Hazard assessment of Maerua subcordata (Gilg) DeWolf. for selected endpoints using a battery of in vitro tests.

ETHNOPHARMACOLOGICAL RELEVANCE: Maerua subcordata (Gilg) DeWolf is a medicinal and wild food plant growing mainly in east Africa. Especially its root tuber is widely used in traditional medicine to treat several infectious and chronic diseases but also in some toxicity implications like use as abortifacient. AIM OF THE STUDY: the present study applied in silico and in vitro tests to identify possible hazards of M. subcordata (fruit, leaf, root, seed) methanol extracts focussing on developmental toxicity. MATERIALS AND METHODS: Ames test, estrogen receptor alpha (ERα) assay, aryl hydrocarbon receptor (AhR) assay, embryonic stem cell test (EST), and zebrafish embryotoxicity test (ZET) were employed. Besides, a Derek Nexus toxicity prediction was performed on candidate structures obtained from metabolomics profiling of the extracts using liquid chromatography coupled to multistage mass spectroscopy (LC/MSn) and a MAGMa software based structural annotation. RESULTS: Glucosinolates, which degrade to isothiocyanates, and biogenic amines were among the candidate molecules identified in the extracts by LC/MSn - MAGMa software structural annotation. Isothiocyanates and some other candidate molecules suggested a positive mutagenicity alert in Derek toxicity predictions. All the extracts showed negative mutagenicity in the Ames test. However, the Derek predictions also identified endocrine and developmental toxicity as possible endpoints of concern. This was further assessed using in vitro tests. Results obtained reveal that leaf extract shows AhR and ERα agonist activities, inhibited differentiation of ES-D3 stem cells into contracting cardiomyocytes in the EST (p < 0.001) as well as inhibited hatching (p < 0.01) and showed acute toxicity (p < 0.01) in the ZET. Also, the fruit extract showed toxicity (p < 0.05) towards zebrafish embryos and both fruit and seed extracts showed AhR agonist activities while root extract was devoid of activity in all in vitro assays. CONCLUSION: The leaf extract tests positive in in vitro tests that may point towards a developmental toxicity hazard. The current evaluations did not raise concerns of genotoxicity or developmental toxicity for the fruit, seed and root extracts. This is important given the use of especially these parts of M. subcordata, in traditional medicine and/or as (famine) food.

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.

Effects of Maerua subcordata (Gilg) DeWolf on electrophile-responsive element (EpRE)-mediated gene expression in vitro.

Plant extracts and phytochemicals may prevent chronic diseases via activation of adaptive cellular stress response pathways including induction of antioxidant and phase II detoxifying enzymes. The regulatory regions of these inducible genes encode the electrophile-response element (EpRE). This study tested the EpRE induction ability of Maerua subcordata (fruit, leaf, root, seed) methanol extracts and selected candidate constituents thereof, identified by liquid chromatography coupled with multistage mass spectroscopy, employing an EpRE luciferase reporter gene assay using hepa-1c1c7 mouse hepatoma cells. A parallel Cytotox CALUX assay using human osteosarcoma U2OS cells was used to monitor any non-specific changes in luciferase activity or cytotoxicity. Results showed that fruit, root, and seed extracts were non-cytotoxic up to a concentration of 30 gram dry weight per litre but the leaf extract exhibited some cytotoxicity and that the leaf (despite some cytotoxicity), fruit, and seed extracts showed strong induction of EpRE mediated gene expression while induction by the root extract was minimal. Selected candidates included glucosinolates, isothiocyanates, and some biogenic amines. Subsequent studies showed that methyl-, ethyl-, isopropyl-, isobutyl- isothiocyanates, and sec-butyl thiocyanate as well as glucobrassicin induced concentration (1-100 µM) dependent EpRE-mediated gene expression while the biogenic amines stachydrine and trigonelline acted as inhibitors of EpRE-mediated gene expression at 100 µM. The identification of glucolepidiin, glucobrassicin, glucocapparin, stachydrine, and trigonelline in all extracts was confirmed using standards and based on multiple reaction monitoring; yet, glucobrassicin level in the root extract was negligible. In conclusion, this study provided a first report on EpRE mediated gene expression effects of M. subcordata; and despite detection of different glucosinolates in all extracts, those containing glucobrassicin particularly displayed high EpRE induction. Because EpRE inducers are cytoprotective and potential chemopreventive agents while inhibitors are suggested adjuvants of chemotherapy, results of this study imply that process manipulation of this plant may result in herbal preparations that may be used as chemopreventive agents or adjuvants of chemotherapies.