Abstract
Aim: The objective of this in-vitro study involves evaluating the protective action of the extracts of L. amara (LA) (whole fruits including seeds) and R. emodi (RE) (rhizomes) at various concentrations on isolated primary rat hepatocytes.
 Methods: The pulverised dried whole fruits of L. amara (LA) and rhizomes of R.emodi (RE) were extracted successively with petroleum ether (PE), ethanol (EE) and distilled water (AE) and vacuum dried. These extracts of LA petroleum ether (PE), ethanolic (EE) and aqueous (AE) extracts and RE obtained were subjected to in vitro studies at doses of 25, 75, 100, and 150 µg/ml and silymarin (250 µg/ml) in CCl4 (1%) intoxicated primary hepatocytes monolayer cultures the hepatoprotective action of all the extracts of both plants at different doses was carried out using isolated rat hepatocytes which were subjected to CCl4 intoxication followed by estimating/ measuring the changes in serum biochemical markers – SGPT, SGOT, ALP, Total proteins (TP), total bilirubin (TB), albumin (ALB) and triglycerides (TGL).
 Results: Hepatoprotective activity against CCl4 was demonstrated in the rat primary monolayer hepatocyte culture using MMT assay with the ethanolic extracts of both plants showing more hepatocyte protective action compared to the aqueous and petroleum ether extracts by reducing the elevated serum marker levels. Alcoholic and aqueous extracts were found to express more protective action towards CCl4 intoxicated isolated primary rat hepatocytes in a dose dependant manner.
 Conclusion: Based on the result, it is suggested that the extract with the most hepatocyte protective action at a dose of 150µg is LA ethanolic extract (viability=88.24%), followed by LA aqueous extract (viability=84.31%), RE ethanolic extract (viability=88.24%) and RE aqueous extract (viability=88.24%) - which are comparable to the reference silymarin with viability at 92.15%. the petroleum ether extracts of both plants showed least hepatic cell viability with LA pet ether extract at 49.02% and RE pet ether extract at 47.85%
Highlights
Liver being a vital organ for detoxification, can be injured or suffer functional impairment leading many health disorders and diseases [1]
Based on the result, it is suggested that the extract with the most hepatocyte protective action at a dose of 150μg is L. amara (LA) ethanolic extract, followed by LA aqueous extract, RE ethanolic extract and RE aqueous extract - which are comparable to the reference silymarin with viability at 92.15%. the petroleum ether extracts of both plants showed least hepatic cell viability with LA pet ether extract at 49.02% and RE pet ether extract at 47.85%
As a consequence of many allopathic drug therapies, such as antitubercular drugs, chronic liver diseases like chronic hepatitis, cirrhosis, including vascular lesions and fatty liver get precipitated such that they often resemble viral hepatitis. It is imperative for the chemically induced hepatic injury to be severely modifying or damaging to liver functions, as sequential hepatic injury is related to the type hepatotoxins used on specific animal species for consequential liver disturbances clinically resembling acute hepatotoxicity, like acute viral hepatitis [2,3]. Such hepatotoxicity has been observed with CCl4 – a most commonly used toxin in experimental studies, as it readily available in pure form, undergoes hepatic metabolism by cytochrome P450 causing free trichloromethyl (CCl3 ) radicals generation, resulting in direct or indirect in damage to membrane and intracellular lipids, nucleic acids, and other molecules which further leads to toxic hepatic damage with the severity ranging from necrosis leading to triacylglycerol accumulation, cirrhosis and cancer depending upon its mode of application and dose [4,5]
Summary
Liver being a vital organ for detoxification, can be injured or suffer functional impairment leading many health disorders and diseases [1]. It is imperative for the chemically induced hepatic injury to be severely modifying or damaging to liver functions, as sequential hepatic injury is related to the type hepatotoxins used on specific animal species for consequential liver disturbances clinically resembling acute hepatotoxicity, like acute viral hepatitis [2,3] Such hepatotoxicity has been observed with CCl4 – a most commonly used toxin in experimental studies, as it readily available in pure form, undergoes hepatic metabolism by cytochrome P450 (especially by P450 2E1) causing free trichloromethyl (CCl3 ) radicals generation, resulting in direct or indirect in damage to membrane and intracellular lipids, nucleic acids, and other molecules which further leads to toxic hepatic damage with the severity ranging from necrosis leading to triacylglycerol accumulation, cirrhosis and cancer depending upon its mode of application and dose [4,5]. Smaller doses in the range of tenths of ml CCl4/kg body weight repeated several times weekly can produce cirrhosis within a few months [7]
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