Study On The Effect And Mechanism Of Extract From Rhus Chinensis Fruit On Preventing Chemical Liver Damag

In this paper,Chinese sumac fruits(Rhus chinensis Mill.)as the experimental material,and 80% ethanol was used to extract the fruit to obtain the extractive.Then,the chemical substances in the extractive of the fruit were quantitatively and qualitatively analyzed by UHPLC-ESI-MS/MS.Meanwhile,the bioactivity of the extract on preventing drug-induced liver injury(DILI)and cholestasis liver injury and liver fibrosis were studied,and further discussion the potential mechanism via western blot.The main results of this paper are as follows:(1)A total of 17 phenolic compounds were identified from the ethanol extract of Chinese sumac fruit,among which the contents of quercetin-3-O-Rhamnoside and gallic acid were the highest.In the experiment of preventing acute liver injury induced by acetaminophen(paracetamol,APAP),the ethanol extract of Chinese sumac fruit could significantly improve the biochemical indexes of ALT,AST,GSH,SOD,MDA and CAT.The results of pathological section showed that the extractive could significantly improve the nuclear atrophy in hepatic lobule and reduce the destruction of cell structure.The ethanol extract of the fruit could significantly regulate the related protein of oxidative stress,such as Nrf2、HO-1 and NQO1.CYP2E1 is a key rate-limiting enzyme in liver injury induced by APAP.It was found that the extractive of the fruit could down-regulate the expression of CYP2E1 to reduce the production of NAPQI.In addition,the extract also inhibits macrophage secretion of pro-inflammatory cytokines(TNF-α,IL-6 and IL-1β),down-regulated the expression of MAPK and NF-κB related protein,thereby inhibiting APAP-induced inflammation.Finally,the results of apoptosis-associated proteins(Bax、Bcl-2 and Caspase3)and TUNEL fluorescence showed that the extract could significantly inhibit apoptosis,and 800 mg/kg b.w.dose of extract has better protective effect.(2)In the experiment on the protective effect of Chinese sumac fruits against liver injury caused by anti-tuberculosis drugs(INH and RIF),the results showed that the ethanol extract of the fruits could significantly improve biochemical indexes such as ALT,AST,ALP,TBIL,SOD,GSH,MDA and CAT.Meanwhile,the extract can inhibit oxidative stress by up-regulating the expression of Nrf2,HO-1 and NQO1.Moreover,the extract reduces the release of proinflammatory factors(TNF-α,IL-6,and IL-1β)to reduce inflammation.WB and TUNEL fluorescence detection showed that the extract could inhibit apoptosis by regulating the expression of apoptosis proteins(Bax and Bcl-2).The extract also slows down the metabolism of INH by down-regulating the expression of CYP2E1.RIF can also cause an increase in bilirubin,which causes bile to accumulate in the liver.The liver excretes bile components,such as bile acids,into the bile ducts primarily through transport proteins BSEP and Mrp2.The expression of the two proteins was detected by WB,and it was found that the ethanol extract of the fruit could regulate the expression of transporters to reduce liver injury.(3)In the experiment of preventing cholestatic liver injury and fibrosis in Chinese sumac fruit,the results showed that the ethanol extract of fruit could significantly improve the biochemical indexes such as ALT AST,ALP,TBIL,SOD,GSH and MDA.The results of H&E Pathological sections showed that ethanol extract of the fruit could reduce the number of necrotic cells around hepatic sinuses and inhibit inflammatory cell infiltration.The results of ELISA and WB found that the ethanol extract of fruit could reduce the secretion of proinflammatory factors(TNF-α,IL-6 and IL-1β)and down-regulate the expression of p-NF-κB and p-IκB,thus alleviating the inflammatory response.In terms of bile acid transport,the ethanol extract of this fruit can regulate the expression of bile acid transport key proteins BSEP and Mrp2 to reduce the accumulate of bile acid.From the perspective of liver fibrosis,Masson staining and Sirius red staining showed that the extract could reduce collagen deposition.Further determination of TGF-β,α-SMA protein expression showed that the ethanol extract could reduce the degree of fibrosis.