The Effects Of Splenenctomy On The Expression Of TGF-β1,bFGF And α-SMA In The Rats With Hepatic Fibrosis

Objective:.To dynamically observes the expressions of TGF-β1,b FGF and α-SMA within rat liver tissue, and discusses the function and mechanism of TGF-β1,b FGF during the process of hepatic fibrosis through establishing rat hepatic fibrosis model; observes the changes in expressions of TGF-β1,b FGF and α-SMA within rat liver and discusses the relations between the development mechanisms of spleen and hepatic fibrosis, by way of cutting off the spleen of the rats at the early, middle and later periods of hepatic fibrosis.Methods:Experiment I: according to the random principle, divide 50 male SD rats into normal group, groups of 2 weeks modeling, 4 weeks, 6 weeks and 8 weeks, and inject 40% carbon tetrachloride solution into the rat peritoneal, with a dose of 2ml/kg, twice a week, totally 8 weeks, establish haptic fibrosis model; the normal group is killed before modeling, and the others are killed when the modeling is 2,4,6and 8 weeks respectively. Experiment II: according to the random principle, divide 48 male SD rats into A,B and C three groups, inject 40% carbon tetrachloride solution into the rat of model group, with a dose of 2ml/kg, twice a week, totally 8 weeks, establish hepatic fibrosis model; establish test group and control group for each big group, cut off the spleen of the rat in A,B,C test groups after 2,4 and 8 weeks of modeling respectively, meanwhile, excise sham operation for each control group; and kill all rats after 8 weeks of modeling.Keep the liver sample and stain HE to observe the pathological change in liver tissue, and detect the expression of TGF-β1,b FGF and α-SMA within rat liver tissue in immunohistochemical methoc; apply Realtime-PCR to detect the level of TGF-β1m RNA, b FGFm RNA and α-SMAm RNA within the rat liver tissue of each group.Results:Experiment 1:(1)The result of immunohistochemical staining shows: as the prolong of modeling time, the expressions of TGF-β1,b FGF and α-SMA within the liver tissue of each model group increase gradually. The TGF-β1,b FGF and α-SMA of the liver tissue of model group are all higher than those of the normal group(P<0.01). The expressions of TGF-β1,b FGF and α-SMA are positive correlation.(2)The result of the Quantitative Real-time PCR shows: as the prolong of modeling time, the levels of TGF-β1m RNA,b FGFm RNA and α-SMAm RNA increase gradually. The levels of TGF-β1m RNA, b FGFm RNA and α-SMAm RNA within the hepar of model group are all higher than those of the normal control group(P<0.01).Experiment 2:(1)The result of immunohistochemical staining shows that the expressions of TGF-β1,b FGF and α-SMA within the liver tissue of each test group are lower than those of the control group(P<0.05). The three test groups of A,B,C make single factor variance and analyze SNK to detect and make comparisons for each pair of mean values, the result shows if the spleen is cut out at the early period of hepatic fibrosis, the expressions of TGF-β1,b FGF and α-SMA within the liver tissue are at the lowest(P<0.05).(2) The result of Quantitative Real-time PCR shows the levels of TGF-β1m RNA,b FGFm RNA and α-SMAm RNA of liver tissue of each test group are lower than those of the control group(P<0.05), The three test groups of A,B,C make singl e factor variance and analyze SNK to detect and compare each pair of mean val ues, the result shows if the spleen excision is cut off at the early period of hep atic fibrosis, the expressions of TGF-β1m RNA,b FGFm RNA and α-SMAm RNA within the liver tissue are at the lowest(P<0.05).Conclusion:1.With the development of hepatic fibrosis, the expressions of TGF-β1,b FGF and α-SMA and levels of TGF-β1m RNA,b FGFm RNA and α-SMAm RNA in liver tissue of rats increase gradually.2.Splenectomy can obviously reduce the expressions of TGF-β1,b FGF and α-SMA within liver tissue, and delay the development of hepatic fibrosis. The spleen has played an important role in the development of hepatic fibrosis.3.Cutting off spleen during the early period of hepatic fibrosis is more effective than cutting off in the middle and later periods of prolonging the development of hepatic fibrosis.