Metformin Inhibits The Development Of Cirrhosis By Regulating The Expression Of KLF4

Cirrhosis is the ultimate stage of liver injury caused by chronic liver disease and various causes.Cirrhosis causes about 1 million deaths a year worldwide,accounting for 2 percent of all deaths,of which 750,000 deaths are due to liver cancer.Due to the large population base of China and the big country of hepatitis B,the incidence of cirrhosis is the highest in the world.At the same time,liver cirrhosis is not only a serious health threat to the world but also a heavy burden on the national economy.Cirrhosis is still considered by many doctors as the end-stage development stage of liver disease,and there is no effective treatment alternative to liver transplantation.However,as the study goes on to find out that cirrhosis is believed to be the development stage of end-stage liver disease,this concept is constantly being challenged.The data found that there is still a different stage of development in the so-called clinical definition of cirrhosis,and the prognosis is completely different in different stages.Therefore,the survival of the prognosis of different stages of liver cirrhosis and its potential reversibility excited researchers to seek treatment strategies for delaying or preventing the progression of cirrhosis.At the same time explore cirrhosis development process for screening of differentially expressed genes provides a research direction,at early stages of cirrhosis of the exploration of new drugs to stop the progress of the cirrhosis of the liver cirrhosis treatment provides a new way.Part ? Screening the aberrantly expressed genes in liver fibrosis of the mouseAimsScreening the aberrantly expressed genes in liver fibrosis of the mouse.MethodsFive cases of liver tissue and control group of liver fibrosis and the isolated and identified rat hepatic stellate cells which were cultured for 1 day,7 days and 14 days were collected and RNA extraction and purification,microarray experiments and data analysis was finished by SHANGHAI BIOTECHNOLOGY CORPORATION.RusultsUse Fold Change(linear)>= 2 and the t-test(Student's t-test)p-value<0.05 for the differential gene screening,all differential expressed genes in the liver fibrosis tissues and the control group are 739,in which overexpressed is 551,and low expressed is 188.In mice,hepatic stellate cells were cultured for 7 days compared with 1 days,the high expression is 699 and the low expression is 495.There were 1784 high expression of hepatic stellate cell culture in mice and 3,555 low expression.In mice,the hepatic stellate cells were cultured for 14 days,compared with those in the 7 days of cultivation,with 143 high expression and 641 low expression.The mouse hepatic stellate cell were cultured for 1 day,7 days,and 14 days,with 25 high expressions and 28 low expression levels.The mouse hepatic stellate cell were cultured for 1 day,7 days,and 14 days compared with the liver fibrosis tissues,with 3 high expressions(Colla2?Fam69b?Gpbarl)and 2 low expression(Ear3?Ednl).ConclusionThe key genes for regulating liver fibrosis in mice were screened by gene chip,which provided the direction for the follow-up mechanism research.Part ? Metformin inhibits the development of liver fibrosis by raising KLF4 expressionAimsTo explore the specific mechanism of metformin in inhibiting the development of liver fibrosis.MethodsSD rat liver fibrosis model was constructed which contains the olive oil group,tetrachloride group,tetrachloride + metformin,tetrachloride + KLF4 group and then HE staining,Massion staining,KLF4 immunohistochemistry,a-SMA immunohistochemistry and compare the expression levels of HE and Massion in dark light and KLF4 and a-SMA.The original hepatic stellate cells of SD rats was isolation,culture and identified.The expression of KLF4 and a-sma were detected in the primary hepatic stellate cells and in the humanized hepatic stellate cells,respectively,with the stimulation of metformin and the control of PBS stimulation and transfection of the KLF4 and the control adenovirus.The PBS group,metformin,metformin,metformin and Si-KLF4 were set up in the primary hepatic stellate cells and the humanized hepatic stellate cells,respectively,and the expression of KLF4 and a-SMA were detected in WB respectively.ResultsIn the immunohistochemical staining of KLF4S the expression of KLF4 in the tetrachloride group was lower than that in the olive oil group,the tetrachloride +metformin group and the tetrachloride + KLF4 group were all higher than the tetrachloride group.In the a-SMA immunohistochemical stain,the a-SMA was expressed in the tetrachloride group higher than the olive oil group,the tetrachloride +dimetformin and the tetrachloride + KLF4 group was lower than the tetrachloride group:In his and Massion staining,the tetrachloride groups were higher than those in the olive oil group,the tetrachloride + metformin and the tetrachloride + KLF4 group were lower than the tetrachloride group.The isolated and cultured SD rats were identified by Desmin immunofluorescence staining to be the original hepatocytes.In the primary hepatic stellate cells and the humanized hepatic stellate cells,metformin was stimulated and overexpressed in the KLF4 group compared with the PBS group and the adenovirus control group,the expression of a-SMA decreased and KLF4 expression increased.In the original generation of hepatic stellate cells and hepatic stellate cells,the humanized KLF4 expression of metformin group is higher than the PBS group,metformin + Si-KLF4 below the metformin group,the expression of a-SMA metformin group is lower than the PBS group,metformin + Si-KLF4 group was higher than metformin group.ConclusionMetformin inhibited the activation of hepatic stellate cells by raising KLF4 expression,thereby slowing the development of liver fibrosis.