| Objective:To observe the effects and the influencing factors of IFNa-2a to liver fibrosis induced by CC14; and to discuss the mechanism of apoptosis of IFNa-2a blocking the liver fibrosis induced by CC14.Method:To establish the model of the liver fibrosis induced by CC14, we used the principles of randomized comparison and divided SD rates into 5 groups, in which each group contained 10, namely the NS control group (A group), fibrosis model group (B group),60,000 U/Kg IFNa-2a treatment group (C group),120,000 U/Kg IFNa-2a treatment group (D group) and the 60,000 U/Kg IFNa-2a control group (E group). After 8 weeks'model established, we collected blood samples and liver tissue samples, and checked liver function ALT, AST, TBIL, TP indicators, the liver fibrosis HA, LN, PCIII indicators, HE staining, masson staining and reticular fiber staining respectively. We adopted semi-quantitative RT-PCR to analyze liver Bcl-2, Bax gene expression, adopted TUNEL to check the apoptosis of liver cells and adopted immunohistochemistry through a-SMA to mark the active hepatic stellate cells. The experiment data are processed by SPSS 17.0 statistical software using analysis of variance.Results:(1) serological tests:the liver function ALT, AST, TBIL between A, B, C, D group were of significant differences (P<0.05), the B group was significantly higher than that of A group,the C, D group were significantly lower than the B group (P<0.05), the D group was significantly lower than that of C group (P<0.05), there was no significant difference between control group A and E group (P> 0.05); Fiber index of liver HA:B group was significantly higher than that of A group, B group was lower than C group (P<0.05), there was no significant difference between D group, A groupand E group groups (P> 0.05); there was no difference of TP among all groups, (P> 0.05). for the LN, there were significantly differences between each experimental group (P<0.05); Histopathology:HE staining, masson staining and reticular fiber staining all showed the degree of liver inflammation and fibrosis of C and D group were significantly reduced than B group,the D group of liver fibrosis are lighter than C group, the A group and E group have no liver inflammation and fibrosis.(2) Semi-quantitative RT-PCR analysis showed that for Bcl-2 gene expression, there were differences between the groups of mean, The expression for the B group (0.827±0.001) is the highest, E group (0.785±0.001) is the lowest, the C group (0.813±0.002), D group (0.796±0.001) and A group (0.790±0.001) between the B, E groups.there were differences among each groups in Bax gene expression, its expression sequence was reversed to the expression of Bcl-2 expression, and they were respectively A group (1.060±0.003), D group (0.916±0.002), C group (0.899±0.003), B group (0.839±0.003),E group (1.055±0.001). TUNEL detection of apoptosis:Liver cells apoptotic index in each experimental group were significantly different (P<0.05), they were respectively:A group (6.98±1.19)%, B group (57.53±2.21)%, C group (39.56±1.17)%, D group (24.76±0.96)%, E group (8.61±1.14)%, IFNα-2a treatment group (C, D group) of the hepatocyte apoptosis index was significantly lower than the fibrosis model group (B group) (P<0.05), the control group (A, E group) was significantly lower than IFNα-2a intervention group (C, D group), and IFNα-2a treatment group D group was significantly lower than C group (P<0.05). the immunohistochemistry of the activated hepatic stellate cells Marked by a-SMA:the liver fibrosis model group (B) have a large number of a-SMA expression, IFNα-2a treatment group C a-SMA expression was decreased compared with group B, D group was much less,the A,E group did not contain any a-SMA expression.Conclusions:(1) IFNα-2a can block the CC14 induced liver fibrosis, its effects were positively correlated with IFNα-2a dose. (2) The mechanism of IFNα-2a blocking the CC14 induced liver fibrosis by regulating Bcl-2, Bax expressionthe regulation induced apoptosis of achieved hepatic stellate cells,The adjustment function may be associated with IFNα-2a dose.(3) IFNα-2a blocks the CC14 induced liver fibrosis possibly to exist the liver cell apoptosis pathway. |
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