Effects Of A 54° Edible Liquor On Hepatocyte Morphology,Apoptosis Index,Expression Of Apoptosis Factor And Metallothionein In Rat Hepatocyte

Objective: To explore effects of a 54° edible liquor on histopathology,ultrastructure of hepatocyte,apoptosis index,expression of cytokines of apoptosis including Bcl-2,Bax,TNF-?,Caspase-9,Caspase-12 and Metallothionein in rats,at the dose and course of this experiment,by gavage of this brand 54°consumption of liquor to SD rats that simulation of normal drinking habits,so as to provide the basic experimental data for the healthy drinking.Methods:(1)Drinking animal model established: 115 normal male SD rats were randomly divided into normal control group(CG,n=25)and experimental group(EG,n=90).The experimental group was divided into low dose group of 0.8ml · kg-1 · d-1(LG),middle dose group of 1.6ml ·kg-1 · d-1(MG),high dose group of 2.4 ml · kg-1 · d-1(HG).The rats of the EG were given by gavage twice a day(11:00 AM an 5:00 PM)with liquor,while rats in CG received no special treatment.Liver were excised at the end of 4 weeks,8weeks,12 weeks,16weeks,and 20 weeks.(2)HE staining technique was used to observe the tissue structure and cell morphology in groups in liver.(3)The ultrastructure of hepatocytes in groups at 4 weeks,8weeks,12 weeks was observed by transmission electron microscopy.(4)TUNEL method was used to detect the apoptosis index of hepatocytes on rats.(5)The expressions of Bcl-2,Bax,in hepatocyte were detected by immunohistochemical SABC,image analysis and morphometric measurement and western blotting in groups in groups.(6)Expressions of Caspase-9 and Caspase-12 in hepatocyte were studied by immunohistochemical SABC,image analysis and morphometric measurement in groups.(7)Expressions of TNF-? in hepatocyte were studied by immunohistochemical SABC,image analysis and morphometric measurement in groups.(8)The expressions of MT in hepatocyte were detected byimmunohistochemical SABC,image analysis,morphometric measurement and western blotting in groups in groups.(9)The one-way ANOVA was used to analyze the experimental data.Results:(1)There were no obvious pathological changes in liver tissue structure and cell morphology in groups showed by HE staining technique;(2)The results of transmission electron microscopy showed that the ultrastructure of hepatocyte in groups at 4 weeks,8 weeks and 12 weeks were not changed.(3)TUNEL method show the positive product of hepatocyte apoptosis exist in the nucleus of the liver in groups and apoptosis index in each dose group was not significantly different from that in normal control group(P>0.05).(4)Bcl-2 and Bax immunohistochemical positive products exist in liver cells cytoplasm in groups;Mean optical density of Bcl-2 and Bax in EG was not significantly different from that in CG(P>0.05).The expression level of Bcl-2 and Bax in the each EG were not significantly different form CG(P>0.05)show by Werstern blotting.(5)Caspase-9immunohistochemical positive products exist in cytoplasm and nucleus of hepatocytes in groups and Caspase-12 in cytoplasm of hepatocytes.The mean optical density of Caspase-9 and Caspase-12 in EG was not significantly different from that in CG(P>0.05),except for 4 weeks MG,12 weeks HG and 20 weeks HG of Caspase-12 increased slightly(P<0.05).(6)The positive products of TNF-? were found in the cytoplasm of hepatocytes in groups.The mean optical density of TNF-? in each dose group was not significantly different from that in CG(P>0.05);(7)Positive products of MT were found in the cytoplasm of hepatocytes in groups.The mean optical density of MT in each dose group was not significantly different from that in CG(P>0.05);Conclusion: In the dose and time course of this experiment,the administration by gavage of this brand 54° consumption of liquor has no effect on the histopathology,ultrastructure,apoptosis index,expression of cytokines of apoptosis including Bcl-2,Bax,TNF-?,Caspase-9,Caspase-12 and MT in rat hepatocyte.