Pilea umbrosa ameliorate CCl induced hepatic injuries by regulating endoplasmic reticulum stress, pro-inflammatory and fibrosis genes in rat

BACKGROUND@#Pilea umbrosa (Urticaceae) is used by local communities (district Abbotabad) for liver disorders, as anticancer, in rheumatism and in skin disorders.@*METHODS@#Methanol extract of P. umbrosa (PUM) was investigated for the presence of polyphenolic constituents by HPLC-DAD analysis. PUM (150 mg/kg and 300 mg/kg) was administered on alternate days for eight weeks in rats exposed with carbon tetrachloride (CCl). Serum analysis was performed for liver function tests while in liver tissues level of antioxidant enzymes and biochemical markers were also studied. In addition, semi quantitative estimation of antioxidant genes, endoplasmic reticulum (ER) induced stress markers, pro-inflammatory cytokines and fibrosis related genes were carried out on liver tissues by RT-PCR analysis. Liver tissues were also studied for histopathological injuries.@*RESULTS@#Level of antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD), peroxidase (POD) and glutathione (GSH) decreased (p < 0.05) whereas level of thiobarbituric acid reactive substance (TBARS), HO and nitrite increased in liver tissues of CCl treated rat. Likewise increase in the level of serum markers; alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP) and total bilirubin was observed. Moreover, CCl caused many fold increase in expression of ER stress markers; glucose regulated protein (GRP-78), x-box binding protein1-total (XBP-1 t), x-box binding protein1-unspliced (XBP-1 u) and x-box binding protein1-spliced (XBP-1 s). The level of inflammatory mediators such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1) was aggregated whereas suppressed the level of antioxidant enzymes; γ-glutamylcysteine ligase (GCLC), protein disulfide isomerase (PDI) and nuclear erythroid 2 p45-related factor 2 (Nrf-2). Additionally, level of fibrosis markers; transforming growth factor-β (TGF-β), Smad-3 and collagen type 1 (Col1-α) increased with CCl induced liver toxicity. Histopathological scrutiny depicted damaged liver cells, neutrophils infiltration and dilated sinusoids in CCl intoxicated rats. PUM was enriched with rutin, catechin, caffeic acid and apigenin as evidenced by HPLC analysis. Simultaneous administration of PUM and CCl in rats retrieved the normal expression of these markers and prevented hepatic injuries.@*CONCLUSION@#Collectively these results suggest that PUM constituted of strong antioxidant chemicals and could be a potential therapeutic agent for stress related liver disorders.

In vitro antioxidant and anti-cancer activities and phytochemical analysis of Commelina benghalensis L. root extracts

Objective: To explore antioxidant potential, anti-cancer activity, and phytochemicals of Commelina benghalensis L. Methods: The roots of Commelina benghalensis were extracted in different solvents (methanol, ethanol, benzene, chloroform, n-hexane) with a range of polarity. Antioxidant activity was evaluated by reducing power assay, DPPH radical scavenging activity and phosphomolybdenum method, cytotoxicity by MTT assay, apoptotic and cell cycle analysis by flow cytometry, migratory and invasive potential by wound scratch assay and invasion assay, respectively, functional groups analysis by FT-IR spectroscopy and phytochemicals by aluminum chloride colorimetric and Folin-Ciocalteu methods. Results: The extracts showed worthy antioxidant potential. The chloroform extract demonstrated the most significant cytotoxic effect on MDA-MB-231 (breast cancer) cell line, induced apoptosis and reduced migratory and invasive potential of MDA-MB-231 cells. Methanol and ethanol extracts presented good yield of total phenolic and total flavonoid contents. The FTIR spectroscopic studies revealed different characteristic peak values with various functional compounds such as alkenes, alkanes, aliphatic amines, aromatics, alkyl halides, carboxylic acid, alcohols, ester, aldehydes and ketones. Conclusions: The results demonstrate the potential use of Commelina benghalensis as a good antioxidant with significant anti-cancer effect.

Inhibitory activities of extracts of Rumex dentatus, Commelina benghalensis, Ajuga bracteosa, Ziziphus mauritiana as well as their compounds of gallic acid and emodin against dengue virus

Objective:To investigate the inhibitory effects against dengue virus serotype 2 (DENV-2) by five different fractions (extracted by methanol, ethanol, benzene, chloroform and n-hexane) of Rumex dentatus, Commelina benghalensis, Ajuga bracteosa and Ziziphus mauritiana, as well as their constituents (gallic acid, emodin, and isovanillic acid).Methods:All the samples were tested for cytotoxicity on baby hamster kidney cells by MTT assay and for anti-DENV-2 activity by plaque reduction neutralization assay using two DENV-2 doses (45 and 90 plaque- forming units or PFU).Results:All the samples except isovanillic acid exhibited significant prophylactic effects against DENV-2 infectivity (without cytotoxicity) when administered to cells before infection, but were not effective when given 6 h post-infection. The methanol extract of Rumex dentatus demonstrated the highest antiviral efficacy by inhibiting DENV-2 replication, with ICConclusions:The highly potent activities of the extracts and constituent compounds of these plants against DENV-2 infectivity highlight their potential as targets for further research to identify novel antiviral agents against dengue.

Inhibitory activities of extracts of Rumex dentatus, Commelina benghalensis, Ajuga bracteosa, Ziziphus mauritiana as well as their compounds of gallic acid and emodin against dengue virus

Objective: To investigate the inhibitory effects against dengue virus serotype 2 (DENV-2) by five different fractions (extracted by methanol, ethanol, benzene, chloroform and n-hexane) of Rumex dentatus, Commelina benghalensis, Ajuga bracteosa and Ziziphus mauritiana, as well as their constituents (gallic acid, emodin, and isovanillic acid). Methods: All the samples were tested for cytotoxicity on baby hamster kidney cells by MTT assay and for anti-DENV-2 activity by plaque reduction neutralization assay using two DENV-2 doses (45 and 90 plaque- forming units or PFU). Results: All the samples except isovanillic acid exhibited significant prophylactic effects against DENV-2 infectivity (without cytotoxicity) when administered to cells before infection, but were not effective when given 6 h post-infection. The methanol extract of Rumex dentatus demonstrated the highest antiviral efficacy by inhibiting DENV-2 replication, with IC