The Role Of FGF21 In The Formation And Resolution Of Alcoholic Liver Fibrosis In Rats

Objective:To study the role of FGF21 in the formation and resolution of alcoholic liver fibrosis in rats.Methods:Fifty healthy Sprague-Dawley rats of 6-8 weeks old?180-200g?and growing in a specific pathogen free?SPF?environment,were selected and fed randomly for one week,then randomly divided into a control group?n=10?and aexperimental group?n=40?.The experimental group was fed with common diet,then taken 10%alcohol by intragastric administration and 40%CCl₄peanut oil mixture by intraperitoneal injection to prepare liver fibrosisrat model.The normal control group was fed with common diet and given 0.9%saline by intragastric and intraperitoneal injection.After 4 weeks of modeling,4 rats in the control group and the experimental group were randomly sacrificed to evaluate the modeling situation.6 weeks later,4rats in the experimental group were randomly sacrificed,and after confirming that the liver fibrosis model was successfully established,the animals were intervened ingroups through preliminary experiments.The remaining experimental rats were randomly divided into a model group,a natural recovery group,and a colchicinegroup.The previous control group was fed with common diet and given 0.9%saline for 2 weeks.The natural recovery group was fed with common diet and given 0.9%saline for 2 weeks.Colchicine group was fed with common diet and given colchicine drug intragastric administration for 2 weeks.In the model group,alcohol intragastric administration was stopped and 50%CCl₄peanut oil mixture was intraperitoneally injected to maintain liver fibrosis for 2 weeks.Then all the rats were sacrificed and tested in the 8th week.Methods:?1?.Taking blood from abdominal aorta after sodium pentobarbital anesthesia,inspecting the following indicators with biochemicalanalyzer:the level of serum biochemical fasting plasma glucose?FPG?,alaninetransaminase?ALT?,aspartate transaminase?AST?,alkaline phosphatase?ALP?,total bilirubin?TBIL?,direct bilirubin?DBIL?and indirect bilirubin?IBIL?,totalcholesterol?CHOL?,triglyceride?TG?,high density lipoprotein?HDL-C?,low density lipoprotein?LDL-C?,glycosylated serum protein?GSP?;The insulin?FINS?level was determined by the insulin release kit.?2?.Taking the liver tissue frozen in therefrigerator at-80?.One part of the liver tissue was used to detecte interleukin-6?IL-6?,tumor necrosis factor-??TNF-??,I collagen?COL I?,?-smooth muscleactin??-SMA?,transforming growth factor-?1?TGF-?1?and FGF21 lever byenzyme-linked immunosorbent assay?ELISA?.Using real-time polymerase chain reaction?rt-PCR?to detect liver FGF21 m RNA expression.Another part of livertissues were fixed with 4%paraformaldehyde solution and embedded in paraffin for HE and Masson staining.Results:1?Induce SD rats with alcohol combined with CCl₄,the liver can show typical alcoholic liver fibrosis pathology at 6w;meanwhile,with the prolongedmodeling time,inflammatory factors and liver fiber indicators??-SAM,COLI,TGF-?1,TNF-?and IL-6?increased significantly,and the liver function indexes?ALT and AST?in the blood increased significantly,and the changes in the liver and blood indexes were consistent with the pathological manifestations.2?During the formation of alcoholic liver fibrosis,the change trend of FGF21 in the liver is to increase first and then decrease.In the later stage of alcoholic liver fibrosis,there is insufficient secretion.During the formation of alcoholic liver fibrosis,thecontent of FGF21 in the liver was negatively correlated with the content of?-SAM,COLI,TGF-?1,TNF-?and IL-6?r=-0.884,-0.905,-0.950,-0.816 and-0.856,P<0.01?.3?Colchicine can significantly improve liver pathological changes of alcoholic liver fibrosis rats,significantly reduce the content of?-SAM,COLI,TGF-?1,TNF-?and IL-6 in the liver,inhibit liver inflammation and collagen fibers Hyperplasia;reduce the content of ALT,AST,TBIL,DBIL in the blood,improve liver function;reduce the content of INS in the blood,improve insulin resistance.4?During the intervention of colchicine in rats with alcoholic liver fibrosis,compared with the model group,the FGF21 content in the colchicine group increasedsignificantly[?5.71±2.16?vs?21.49±10.2?pg/ml,?P<0.05?];In addition,the expression level of FGF21m RNA also increased significantly[?0.92±0.11?vs?2.33±0.42?,?P<0.05?].Conclusion:1?Alcohol combined with CCl₄can successfully induce alcoholic liver fibrosis model in SD rats.2?During the formation of alcoholic liver fibrosis,liver FGF21 may play a role in resisting inflammation and HSC activation.Insufficient FGF21 secretion in the liver in the late stage of alcoholic liver fibrosis may be one of the reasons for the progress of liver fibrosis.3?Colchicine can successfully reverse alcoholic liver fibrosis in rats and increase the content and expression level of FGF21 in the liver.It was suggested that the increase of FGF21 in vivo may be the promoter of the reversal of alcoholic liver fibrosis.