The Effect Of Basic Fibroblast Growth Factor In Bone Marrow Mesenchymal Stem Cells On The Treatment Of Liver Cirrhosis In Mice

BACKGROUND:The therapeutic potential of stem cells in liver fibrosis in mice had been studied widely in experiment of animals and clinic.Basic fibroblast growth factor could improved the proliferation and activity of stem cell in vitro,but the therapeutic effect of bFGF treated BMSC in liver cirrhosis is still unclear.OBJECTIVE: To investigate the effect of b FGF on the growth and proliferation of BMSC.Investigating liver function of cirrhotic mice after bone marrow mensenchymal stem cells translation through caudal veins.METHODS:BMSCs were extracted from a male SD rat by whole bone arrow adherent method.Female KM mice were randomly divided into five groups: group A(CCl4+PBS),group B(CCl4+bFGF),group C(CCl4+bFGF-BMSC),group D(CCl4+ one time infusion bFGF+BMSC),group E(CCl4+ three times infusion bFGF+BMSC).bFGF+ or bFGF-BMSCs were infusion through caudal vein after ten weeks through intraperitoneal injection of CCl4.Liver function was texted by ELLISA,hepatic histologic was texted by HE and masson trichrome.RESULT:(1)the BMSC growth condition,observe each groups of BMSCs growth,both groups BMSCs with or without bFGF after 24 h culture completely adherent cells,early stage adherent cells were fusiform or irregular shape and no obvious abnormal cell morphology found,bFGF + group of BMSCs with bFGF proliferation faster,which covered bottle after 4 days and reach the requirements for subculture.BMSCs culture without b FGF cells growth is relatively slow,which almost completely covered with cell culture bottle after 6 to 7 days.(2)The growth rate of BMSC in vitro was significantly higher than that without b FGF in 10ng/ml bFGF culture.(3)Flow cytometry was used to detect BMSC positive expression of CD29 and CD44 in rats.Negative expression CD45.(4)The liver function was detected by chemical method which revealed that the level of ALT in the model group(group A)had no significant difference compared with group B(P>0.05),but ALT level in group A,group B were decreased compared with group C,group D,group E,(P<0.05);the level of ALT in group D and group E were lower than that in group C(P<0.05);in group E was lower than in group D(P<0.05).There was no significant difference in the level of AST between group A and the group B(P>0.05);there was no significant difference between the group C,group D and group E(P>0.05),while group C,group D and the group E had a significant decrease compared with the group A or group B(P<0.05).(5)HE staining results: in group A and group B showed obvious edema,fatty degeneration,bridging necrosis,irregular arrangement of hepatic cords and extensive formation of false lobules.In group C of liver cell edema and fatty degeneration,inflammatory cell infiltration,but the degree was slightly reduced compared with model group,periportal fibrous deposition,visible pseudolobule structure;in group D,group E,liver cell edema,inflammatory cell infiltration was significantly improved compared with those in group C and pseudolobules were visible,periportal fibrous deposition.(6)Masson-trichrome staining result: The results of Masson showed that large numbers of green stained fibers were deposited in the hepatic portal area of the A group and group B,and the green fibrous invasion in the liver parenchyma,and the fine fibrous septa extending from the portal area to the liver parenchyma,separating the hepatic lobules into lobules.Compared with the B group had no significant difference between the area of group A fiber(P>0.05);compared with group A and group B,group C,fibers of group D and group E was decreased significantly than group A,group B,the difference was statistically significant(all P <0.05);in group C,group D,hepatic portal the essence of fibrous deposition,but there was no significant difference between the two groups of fiber area(P>0.05);fiber area in groupE were significantly lower than in group C and group D(P>0.05).CONCLUSIONS: After cultured in vitro,the growth rate of BMSC was significantly higher than that without b FGF,and the cell morphology was approximately the same.The cell surface markers CD29,CD44 and CD45 expressed by flow cytometry were the same as those without bFGF.b FGF+BMSC transplantation can improve liver function,liver cirrhosis liver cirrhosis,collagen deposition,bFGF is added to the medium of BMSC more significantly improved the liver injury and fibrosis,also hepatic fibrosis was more significant improved after repeated cell transplantation.