| ObjectivePre-protective administration of Mice Ficus pandurata Hance extract to mice,to study the protective effect and its effective parts of Ficus pandurata Hance extract on acute alcoholic liver injury,and to study the mechanism of hepatoprotective effect of its effective parts,further development utilization and clinical application provide experimental basis.MethodsPart 1:Study on the effect of water extracts from different parts of Ficus pandurata Hance on acute alcoholic liver injuryFifty six male C57BL mice(20±2g)were randomly divided into normal control group,acute alcoholic liver injury model control group,positive drug(silybin)group,Ficus pandurata Hance root water extract group(48g/kg),Ficus pandurata Hance stem water extract group(48g/kg),Ficus pandurata Hance leaf water extract group(48g/kg),Ficus pandurata Hance tree water extract group(48g/kg).Among them,the positive drug(silybin)group and the extract group of Ficus pandurata Hance in different parts were administrated by gavage for 18 days;the normal group and the model group were administrated by gavage with pure water of equal volume.After 15 days of intragastric administration,mice in model group,silybin group and Ficus pandurata Hance extract group were given 50%ethanol(12mL/kg)to establish acute alcoholic liver injury model.After fasting for 14 hours after the end of the last modeling,blood was taken from the eyeball,the liver was taken and the liver index was calculated;he staining was used to observe the histopathological changes of the liver;the serum ALT,AST and ALP were detected by the kit.Part two:The study on the protective effect and mechanism of water extract from ground part of banyan on acute alcoholic liver injury48 male C57BL mice(20±2g)were randomly divided into blank control group,acute alcoholic liver injury model control group,positive drug(silybin)group,low dose group(12g/kg),medium dose group(24g/kg)and high dose group(48g/kg)according to their weight.Among them,the positive drug(silybin)group and different doses of banyan extract group were given by gavage for 18 days;the normal group and the model group were given by gavage with equal volume of pure water.After 15 days of intragastric administration,mice in model group,positive drug group(silybin)and different doses of Ficus pandurata Hance extract group were given 50%ethanol(12 mL/kg)to establish the acute alcoholic liver injury model.Fasting after the last modeling 14h,the blood was extracted from eyeball,the liver was taken and the liver index was calculated;the active oxygen was detected in situ by frozen section;the histopathological changes of liver were observed by HE staining;the activities of ALT,AST,LDH and TNF-α in serum of mice were detected by the kit;the malondialdehyde in liver was detected by the kit MDA,SOD,GSH,ROS,IL-1β,IL-18,IL-6,NLRP3,caspase-1,Nrf2,keap-1,HO-1,NQ01 and GCLM were detected by Western blot.Results1.Protective effect of water extracts from different parts of banyan on acute alcoholic liver injuryThe results of liver function test kit showed that:compared with the normal group,the liver index ALT,AST and ALP in the model group were significantly increased(P<0.05);compared with the model group,the liver index ALT,AST and ALP in the intervention group were significantly decreased(P<0.05).Among them,the liver index ALT,AST and ALP of stem and leaf water extract group were significantly lower than that of root extract group(P<0.05).He staining showed that compared with the normal group,the hepatocytes in the model group were disordered,the vacuoles in the liver tissue were enlarged,the space of hepatocytes was unclear,the cytoplasm was light stained,edema appeared,and necrotic area was seen;compared with the model group,the liver histopathological damage in the group treated with banyan extract was alleviated in varying degrees.2.Protective effect of extracts from the ground of Ficus pan durata Hance and banyan on acute alcoholic liver injuryLiver function test kit showed that:compared with the normal group,the liver index ALT,AST,LDH in the model group were significantly increased;compared with the model group,the liver index ALT,AST,LDH in the high dose group of Ficus pandurata Hance extract were significantly decreased(P<0.05);the liver index ALT,AST,LDH in the low and medium dose group were decreased(P>0.05).He staining of liver pathological sections showed that:compared with the normal group,the model group had disorder of hepatocyte arrangement,increase of liver tissue vacuole,unclear hepatocyte space,light staining of cytoplasm,edema,and necrosis area;compared with the model group,the high,medium and low dose of Ficus pandurata Hance extract group had different degrees of liver histopathological damage reduction.3.Effects of extracts from the ground part of Ficus pandurata Hance on the content of oxidative stress index in liver tissue.The results showed that:compared with the normal group,the content of ROS and MDA in the model group increased significantly(P<0.05);compared with the model group,the content of ROS and MDA in the high dose group decreased significantly(P<0.05);the content of SOD and GSH in the low and medium dose group decreased significantly(P>0.05);the content of SOD and GSH in the model group decreased significantly(P<0.05).Compared with the model group,the content of SOD and GSH in liver tissue of the high dose group increased significantly(P<0.05);the content of SOD and GSH in the low and medium dose group decreased(P>0.05).The results of in situ detection of ROS showed that:compared with the normal group,the amount of ROS in the liver tissue of the model group increased significantly;compared with the model group,ROS in the liver tissue of the high,medium and low dose group of banyan extract decreased in varying degrees.4.The effect of extracts from the ground part of banyan on the content of inflammatory factors in liver tissueThe results of ELISA showed that:compared with the normal group,the content of IL-18,IL-6 and IL-1β in the liver tissue of the model group increased significantly(P<0.05);compared with the model group,the content of IL-18,IL-6 and IL-1β in the liver tissue of the high dose group of Ficus pandurata Hance extract decreased significantly(P<0.05);the content of IL-18,IL-6 and IL-1β in the low and middle dose group decreased significantly(P>0.05).5.Effects of extracts from Ficus pandurata Hance on the expression of focal protein NLRP3 and Caspase-1 in liver tissue.The results of Western blot showed that the expression of NLRP3 and Caspase-1 in liver tissue of model group was significantly higher than that of normal group(P<0.05);the expression of NLRP3 and Caspase-1 in liver tissue of high dose group of Ficus pandurata Hance extract was significantly lower than the model group(P<0.05);the expression of NLRP3 and Caspase-1 in low and medium dose group was decreased(P>0.05).6.Effects of extracts from Ficus pandurata Hance on the expression of Nrf2,HO-1,NQ01 and GCLM in liver tissueWestern blot compared with the normal group,the expression of Nrf2,HO-1,NQ01 and GCLM in the model group decreased was not pronounced(P>0.05);compared with the model group,the expression of Nrf2,keap-1,HO-1,NQ01 and GCLM in the high dose group increased significantly(P<0.05);the expression of Nrf2,keap-1,HO-1,N Q01 and GCLM in the low and medium dose group increase trend were not pronounced(P>0.05).Conclusion1 The water extracts from different parts of Ficus pandurata Hance have protective effects on acute alcoholic liver injury,and the water extracts from stem and leaf are better than the water extracts from root.2 The mechanism of action may be that HO-1,NQ01 and GCLM antioxidant proteins produced by Keap1-Nrf2/ARE pathway are up-regulated,so as to reduce the production of ROS in the body.The reduction of ROS leads to the decrease of the level of cell scorch,which further inhibits the liver injury caused by acute alcohol. |
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