Effects Of A Transition Of Liver Sinusoidal Endothelial Cells Regulated By Rho GTPase Signaling On Sinusoidal Capillarization With Schistosomal Hepatic Fibrosis Mouses.

PART I Hepatic Sinusoidal Pathology of Hepatic Fibrosis Mouses Induced by Infection of Schistosoma Japonicum.Objective To investigate the hepatic sinusoidal pathology of hepatic fobrosis mouses in duced by infection of Schistosoma japonicum .Methods Liver tissues from 30 hepatic cirrhotic mouses infected by Schistosoma japonicum and 10 normal mouses were removed and observed under light and electronmicroscopy for hepatic sinusoidal pathology. distribution of CD31,FVIIIRAg were detected by imunohistochemistry,.Results Hematoxylin and Eosin (HE) staining indicated schistosome egg aggradation in the hilum and inflammatory cell infilt ration around the schistosome eggs. Masson trichrome staining showed hyperplasia of blue-green fiber connective tissue around Disse spaces mostly. Under transmission electronmicroscopy , it was found there was serious damage of hepatic sinusoidal endothelium cells , collapse of most cells , infilt ration of inflammatory cells in hepatic sinusoidal , expansion of small bile duct , decreased or injured microfloss , and capillarization of hepatic sinusoidal. Compared with control group, CD31,FVIIIRAg protein staining of sinusoidal endothelial cell in schistosomal hepatic fibrosis mouses increased significantly . Conclusion Our results suggest that a transition of liver sinusoidal endothelial cells may be involved with the development of sinusoidal capillarization with schistosomal hepatic fibrosis mouses.PART II Expression of connective tissue growth factor in sinusoidal endothelial cell of schistosomal hepatic fibrosis mouses.Objective to explore the possible correlations with CTGF,liver sinusoidal endothelial cell and the formation of basal membrane under hepatic sinusoidal endothelial cell by observing the expression of connective tissue growth factor(CTGF)in sinusoidal endothelial cell of schistosomal hepatic fibrosis mouses.Methods the liver fibrosis model was established by abdominal infected with schistosomal cercaria in 40 mouse,and the contral group 40 yet.the dynamic changes of liver fibrosis was observed at different time points(45d,60d,90d,120d)by optic and electonmicrocopy.the distribution of CTGF,collagen type IV(ColIV)and laminin(LN)were detected by imunohistochemistry,and adopt quantitive analysis;proteins of CTGF were detected by Western blotting;CTGFmRNAs were detected by RT–PCR.Results Compared with control group, CTGF protein staining of sinusoidal endothelial cell in schistosomal hepatic fibrosis mouses increased significantly at 120d and companied by the increases of LN,ColIV protein in hepatic sinusoidal walls(P < 0. 05)and the expression of CTGFmRNA was significant increased, and the expression of CTGF protein was significant corrected to the level of LN,ColIV(r=0.7512,0.6417 P <0. 01)yet.Conclusin hepatic sinusoidal endothelial cell of schistosomal hepatic fibrosis mouses regulate producing of extracellular matrix by upregulated expression of CTGF which induced the increases of LN,ColIV levels,and involve the formation of basal membrane under hepatic sinusoidal endothelial cell.PART III Effects of a transition of liver sinusoidal endothelial cells regulated by Rho GTPase signaling on sinusoidal capillarization with schistosomal hepatic fibrosis mouses.Objective To investigate Rho GTPase mediated regulation of a transition of liver sinusoidal endothelial cells on sinusoidal capillarization with schistosomal hepatic fibrosis mouses and possible mechanism.Methods The liver fibrosis model was established by abdominal infected with schistosomal cercaria in 88 mouse,treated with Biltricide in 13W and hydroxyfasudil in 14W.And then they were divided into the following 5 groups,i.e. A,contral groups. B,model groups. C,treated with Biltricide groups.D,treated with hydroxyfasudil groups.E,treated with Biltricide+hydroxyfasudil groups.In 13W(A group 10 mouses and B group 6 mouses),in 16W,19W and 21W(B,D,C,E group,each group 6 mouses),the mouses were sacrificed,respectively.The relative area of liver fibrosis on pathological section was semi-quantitatively determined and assessed by HE,Masson stain,electonmicrocopy and imunohistochemistry.And p-moesin,CTGF,Rho A,ColIV and LN protein expressions were assessed by Western blotting,CTGF,Rho A,ROCKII mRNA expressions were assessed by RT–PCR.Results Compared with A group,the mRNA level of Rho A,ROCKII and CTGF were significantly increased(P<0.05)and the protein expression of p-moesin,CTGF,RhoA,ColIV and LN were increased(P<0.05)in B group.After intervension with Biltricide and/or hydroxyfasudil,CTGF mRNA expression significantly decreased(P<0.05)in E group in 16W and the protein expression of CTGF,ColIV and LN were decreased(P<0.05)compared with other groups,and the expression of p-moesin in E group were markedly lower than that of D group(P<0.05).Electonmicrocopy show liver sinusoidal of the mouses in E group was significantly recovered compared with other groups.And there was no difference between B group and D group.Conclusin Our results suggest that an upregulation of Rho GTPase that contributes to increased CTGF expressions and phosphorylation of moesin may induce a transition of liver sinusoidal endothelial cells on sinusoidal capillarization with schistosomal hepatic fibrosis mouses.