干扰素α对乙型肝炎小鼠病理形态学及TGF-β1、ACT-A表达影响

Effects of interferon alpha on pathomorphology and expressions of TGF-β1 and ACT-A in mice with hepatitis B

目的:探究干扰素α(IFN-α)对乙型肝炎小鼠病理形态学及转化生长因子β1(TGF-β1)和活化素A(ACT-A)表达影响。方法:选用雌性BALB/c小鼠30只,采用随机数表法将其分为对照组、模型组和IFN-α组,每组10只。通过尾静脉注射乙型肝炎病毒(HBV)的方法建立乙型肝炎模型,经尾静脉注射p KCMvint.IFN-α-2a促进IFN-α的表达水平。采用酶联免疫吸附法检测血清IFN-α、谷氨酸转移酶(ALT)、门冬氨酸转移酶(AST)、HBV血清标志物HBe Ag、Hbs Ag、肿瘤坏死因子α(TNF-α)以及白细胞介素6(IL-6)。观察HE染色和Masson染色检测肝组织病理学改变和纤维化情况。采用逆转录-聚合酶链反应(RT-Qpcr)和蛋白印迹方法分别检测TGF-β1、ACT-A的m RNA和蛋白的水平。结果:模型组及IFN-α组小鼠血清中可检测出HBV标志物HBe Ag和HBs Ag,且IFN-α组血清HBe Ag和HBs Ag水平显著低于模型组(t=6.132,t=5.986;P<0.05)。IFN-α组的IFN-α水平显著高于模型组(t=4.812,P<0.05)。模型组ALT、AST和炎性因子TNF-α、IL-6显著高于对照组(t=6.115,t=6.548,t=5.771,t=6.238;P<0.05)。IFN-α组的ALT、AST、TNF-α和IL-6水平显著低于对照组(t=4.661,t=5.018,t=5.823,t=5.339;P<0.05)。对照组小鼠的肝组织正常且未发现纤维化现象;模型组小鼠肝组织出现病理学改变及炎症反应和纤维化;IFN-α组肝组织损伤程度较模型组显著缓解,仅有少量的纤维化染色。模型组小鼠肝组织中TGF-β1、ACT-A的m RNA和蛋白水平显著高于对照组(t(TGF-β1)=5.061,t=5.018;t(ACT-A)=4.718,t=4.059;P<0.05)。IFN-α组TGF-β1、ACT-A的m RNA和蛋白水平显著低于模型组(t(TGF-β1)=6.224,t=4.389;t(ACT-A)=5.973,t=5.389;P<0.05)。结论:IFN-α不但可以抑制HBV的复制,还可以通过控制ACT-A、TGF-β1的表达缓解炎性反应,并减少HBV感染引起的肝组织损伤和纤维化。

Objective:To investigate the effects of interferon alpha(IFN-α)on the pathomorphology and the expressions of transforming growth factor(TGF)-β1 and activin A(ACT-A)in mice with hepatitis B.Methods:30 female BALB/c mice were selected and were randomly divided into the control group,the model group and the IFN-αgroup,with 10 mice in each group.Hepatitis B model was established by injecting hepatitis B virus(HBV)into the tail vein.The expression level of IFN-αwas promoted by tail vein injection of p KCMvint.IFN-α-2a.The serum IFN-α,serum glutamate transferase(ALT),aspartate transferase(AST),HBe Ag,Hbs Ag,tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)of HBV serum markers were detected by enzyme-linked immunosorbent assay.Hepatic histopathological changes and fibrosis were detected by HE staining and Masson staining.The levels of m RNA and protein of TGF-β1 and ACT-A were detected by reverse transcription-polymerase chain reaction(RT-Qpcr)and Western blot,respectively.Results:The HBe Ag and HBs Ag of HBV markers could be detected in the serums of model group and IFN-αgroup,and the serum HBe Ag and HBs Ag levels of IFN-αgroup were significantly lower than those of model group(t=6.132,t=5.986,P<0.05).The IFN-αlevel of IFN-αgroup was significantly higher than that of model group(t=4.812,P<0.05).Serum ALT,AST and inflammatory factor TNF-αand IL-6 of liver function indexes of model group were significantly higher than those of control group(t=6.115,t=6.548,t=5.771,t=6.238,P<0.05).The levels of ALT,AST,TNF-αand IL-6 of IFN-αgroup were significantly lower than those of control group(t=4.661,t=5.018,t=5.823,t=5.339,P<0.05).Liver tissues of these mice of control group were normal and the fibrosis phenomenon didn’t be found in them.Pathological changes,inflammatory reaction and fibrosis occurred in liver tissue of these mice of model group.The degree of liver tissue damage of IFN-αgroup was significantly relieved compared with that of model group,and only a small amount of fibrosis staining could be observed.And the levels of m RNA and protein of TGF-β1 and ACT-A in the liver tissues of model group were significantly higher than those of control group(tTGF-β1=5.061,t=5.018,tACT-A=4.718,t=4.059,P<0.05),respectively.And the levels of m RNA and protein of TGF-β1 and ACT-A of IFN-αgroup were significantly lower than those of model group(tTGF-β1=6.224,t=4.389,tACT-A=5.973,t=5.389,P<0.05).Conclusion:IFN-αcan not only inhibit the replication of HBV,but also relieve the inflammatory response by controlling the expression of ACT-A and TGF-β1,and it also can reduce liver tissue damage and fibrosis caused by HBV infection.