| Objective Lipopolysaccharide,which is one of the factors that contribute to the pathogenesis of chronic hepatitis B(CHB),plays an important role in induction of liver inflammation and fibrosis. However,in addition to inflammation and fibrosis, different degree of intrahepatic vascular lesions can also be observed in the liver tissues of CHB. We measured the serum concentrations of LPS(Lipopolysaccharides),TNF-α(Tumor necrosis factor alpha) and sTM(soluble Thrombomodulin) and explored the expression of TLR4,TNF-α and TM multiclonal antibodies in liver tissues in order to assess the effects of LPS on intrahepatic vascular pathologic changes of CHB.Material and Methods120cases of patients with CHB chosen at random were took down liver tissues by liver biopsy.10cases of normal liver tissues were chosen as the control group.Liver tissues were all fixed in10%formaldehyde and enbedded in paraffin by routine methods. Consecutive sections of the speciemens were stained with hematoxylin and eosin stain technique(HE),and then were examined the pathological changes by light microscope.120patients with CHB were classified as grade146cases, grade238cases, grade336cases by intrahepatic vascular pathologic changes.The serum concentrations of LPS,TNF-α and sTM were quantitatively determinated with enzyme linked immunosorbent assay (ELISA). The expression levels and localization of TLR4,TNF-α and TM multiclonal antibodies in liver tissues were determined by immunohistochemical staining(SABC method).The results were evaluated by integrative index.All data were statistically analyzed by use of SPSS version17.0. Result (1) The serum LPS level in the control group was very low (0.30±0.13ng/ml). The serum LPS level in patients with intrahepatic vascular lesion grade1was0.32±0.14ng/ml,showing no signifiant difference compared with the control group (P>0.05). The serum LPS levels of CHB patient with intrahepatic vascular lesion grade2and grade3were0.71±0.14ng/ml and1.10±0.15ng/ml respectivity, which were significantly higher than that of the control group (P<0.01). There was a positive correlation between serum LPS level and the grade of intrahepatic vascular lesion (r=0.892,P<0.01).(2) The serum TNF-a concentrations of CHB patients with intrahepatic vascular lesion grade1,grade2and grade3were26.14±5.01pg/ml,31.10±5.03pg/ml and48.90±5.05pg/ml respectivity, which were significantly higher than that of the control group (P<0.01). There was a positive correlation between serum TNF-a concentration and the grade of intrahepatic vascular lesion(r=0.848,P<0.01). The higher the serum TNF-a concentration was, the severer intrahepatic vascular lesion would be.(3) The serum sTM levels in patient with intrahepatic vascular lesion were significantly higher than those in controls(P<0.01).In patient with CHB,the serum level of sTM was positively correlated with the grade of intrahepatic vascular lesion(r=0.926,P<0.01).(4)The serum LPS level in patient with CHB was positively correlated with the serum level of TNF-a and sTM (r=0.930,0.936,P<0.01).(5)The intensity of TLR4expression in liver tissue of CHB was significantly higher than that of the control group (P<0.01). The positive staining was mainly located on cell membrane and some in cytoplasm.In CHB,the intensity of TLR4was positively correlated with the grade of intrahepatic vascular lesion (r=0.782,P<0.01).(6)The expression of TNF-a in patients with CHB was significantly increased as compared with that in control (P<0.01). The positive staining was mainly located in cytoplasm and some on cell membrane.The positive expression of TNF-a was significantly correlated with the grade of intrahepatic vascular lesion (r=0.634,P<0.01). (7) There was no signifiant difference in the intensities of TM expression between intrahepatic vascular lesion grade1group and control group(P>0.05). However,the intensities of TM expression in liver tissues of hepatic vascular lesion grade2and grade3were significantly higher than those in controls(P<0.05,P<0.01). The positive staining was mainly located in cytoplasm.The positive expression of TM was significantly correlated with the grade of intrahepatic vascular lesion(r=0.575, P<0.01).Conclusions (1) LPS,TLR4and TNF-a play important roles in the course of intrahepatic vascular pathologic changes of CHB.(2)LPS might induce hepatic vascular lesion by up-regulation the expression of TLR4.(3)Serum LPS, TNF-α and sTM can be used as the indexes to evaluate hepatic vascular lesion. |
PDF Full Text Request