Abstract
The progress in industrialization has blessed mankind with a technologically superior lifestyle but poor management of industrial waste has in turn poisoned nature. One such chemical is carbon tetra chloride (CCl4), which is a potent environmental toxin emitted from chemical industries and its presence in the atmosphere is increasing at an alarming rate. Presence of CCl4 in human body is reported to cause liver damage through free radical mediated inflammatory processes. Kupffer cells present in the liver are potentially more sensitive to oxidative stress than hepatocytes. Kuffer cells produced tumor necrosis factor-α (TNF-α) in response to reactive oxygen species (ROS), that might further cause inflammation or apoptosis. In this study hepatoprotective capacity of antioxidant rich extract of Croton bonplandianus Baill. (CBL) was evaluated on CCl4 induced acute hepatotoxicity in murine model. Hydro-methanolic extract of C. bonplandianus leaf was used for evaluation of free radical scavenging activity. Liver cells of experimental mice were damaged using CCl4 and subsequently hepatoprotective potential of the plant extract was evaluated using series of in-vivo and in-vitro studies. In the hepatoprotective study, silymarin was used as a positive control. Antioxidant enzymes, pro-inflammatory markers, liver enzymatic and biochemical parameters were studied to evaluate hepatoprotective activity of Croton bonplandianus leaf extract. Free radical scavenging activity of CBL extract was also observed in WRL-68 cell line. The phytochemicals identified by GCMS analysis were scrutinized using in-silico molecular docking procedure. The results showed that CBL extract have potent free radical scavenging capacity. The biochemical parameters were over expressed due to CCl4 administration, which were significantly normalized by CBL extract treatment. This finding was also supported by histopathological evidences showing less hepatocellularnecrosis, inflammation and fibrosis in CBL and silymarin treated group, compared to CCl4 group. ROS generated due to H2O2 in WRL-68 cell line were normalize in the highest group (200 μg/ml) when compared with control and negative control (CCl4) group. After molecular docking analysis, it was observed that the compound α-amyrin present in the leaf extract of C. bonplandianus has better potentiality to protect hepatocellular damages than the standard drug Silymarin. The present study provided supportive evidence that CBL extract possesses potent hepatoprotective capacity by ameliorating haloalkane induced liver injury in the murine model. The antioxidant and anti-inflammatory activities also affirm the same. The synergistic effects of the phytochemicals present in CBL are to be credited for all the hepatoprotective activity claimed above.
Highlights
In aerobic system, oxygen and nitrogen are abundantly found
Leaf extract of Croton bonplandianus Baill. (CBL) showed lower IC50 value than ascorbic acid, mannitol, sodium pyruvate, curcumin, quercetin in DPPH, hydroxyl radical, H2O2, nitric oxide, superoxide anion, hypochlorous acid scavenging assay and found comparable with peroxinitrate shown by gallic acid and lipid peroxidation shown by trolox
Antioxidant property covers a broad spectrum of chemical phenomenon and definite antioxidant activity should not be concluded based on a single experimental model
Summary
Oxygen and nitrogen are abundantly found. These molecules take part in various physiological and metabolic processes and undergo changes [1]. The terms Reactive Oxygen Species (ROS) and Reactive Nitrogen Species (RNS) were coined to include all these reactive radical and non-radical molecules which are produced by different endogenous and exogenous sources inside living system [4]. ROS and RNS have certain beneficial role like signal transduction, bodies defence mechanisms against microbes etc., and are required in trace amount inside our body. Due to their chemical nature they helps lipid peroxidation causes DNA damage and oxidize several molecules including cellular membrane and causes injury [5]. Nitrosative stress is dangerous and arises mainly due to nitrogen free radical [6]
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