不同病毒载量的乙型肝炎病毒在不同ALT状态下对肝星状细胞增殖、细胞周期及细胞分泌的影响  

Effects of Hepatitis B Virus with Different Viral Loads at Dfferent ALT Levels on Proliferation,Cell Cycle and Cell Secretion of Hepatic Stellate Cells

作　　者：高鹏[1] 刘才周 朱陇东[2] 李俊峰[2] 于海涛[1] 姚立琼[1] GAO Peng;LIU Caizhou;ZHU Longdong;LI Junfeng;YU Haitao;YAO Liqiong(Department of Clinical Laboratory,First Hospital of Lanzhou University,Lanzhou 730000,China;Department of infectious disease,First Hospital of Lanzhou University,Lanzhou 730000,China)

机构地区：[1]兰州大学第一医院医学检验中心,甘肃兰州730000 [2]兰州大学第一医院传染科,甘肃兰州730000

出　　处：《中山大学学报（医学科学版）》2024年第6期1006-1016,共11页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(81800528)。

摘　　要：【目的】临床上经常以ALT是否正常等因素将乙型肝炎感染者分为乙型肝炎携带者或乙型肝炎患者,并且以ALT≥2UNL作为临床抗病毒治疗的适应症之一,但是目前尚缺乏足够的文献阐明其理论学依据,本文借助乙型肝炎病毒对肝星状细胞的影响对此进行探讨。【方法】分层随机选取不同ALT状态下不同病毒载量的慢性乙型肝炎的患者132例,作为干扰血清。经干扰血清干预HSC后,分别用MTT法检测HSC增殖,流式细胞仪检测HSC细胞周期,RT-PCR和免疫印迹分别检测TGF-β1、Smad3、Smad7、α-SMA、collagenⅠ、collagenⅢmRNA及蛋白的表达,同时对ALT≥2UNL的慢性乙肝患者抗病毒治疗随访24周,对比分析治疗前后患者干扰血清对肝星状细胞的影响。【结果】在ALT正常状态下,不同病毒载量的乙型肝炎病毒对肝星状细胞增殖、细胞周期及细胞分泌无差别性影响。而在ALT异常状态下,特别是ALT≥2UNL组中,随着病毒载量的增加,HSC增殖加速,处于G0/G1期的明显减少,而处于S期,G2/M期的明显增多以及TGF-β1、Smad3、CollagenⅠ、CollagenⅢ、α-SMA mRNA及相应蛋白的表达明显增高,Smad7 mRNA及蛋白明显减少,差异有统计学意义。与抗病毒治疗前相比抗病毒治疗后,干扰血清对肝星状细胞的影响明显减弱。【结论】本文揭示了在不同ALT状态下乙型肝炎病毒对HSC的差异性影响,并通过抗病毒治疗前后对比分析。补充说明了以ALT为节点抗病毒治疗乙型肝炎病毒的细胞学理论基础。【Objective】People infected with Hepatitis B are often divided into hepatitis B carriers and hepatitis B patients based on whether ALT is normal or not,and ALT≥2UNL is one of the indications for clinical antiviral treatment,but no sufficient evidence to justify this.In order to explore the theoretical basis,the paper investigated the effects of hepatitis B virus(HBV)on hepatic stellate cells(HSCs).【Methods】A total of 132 chronic hepatitis B patients with different viral loads and ALT levels were randomly selected as the study subjects.Of these patients,those with ALT≥2UNL were treated with antiviral therapy and followed up for 24 weeks.The effects of HBV on HSCs before and after the treatment were compared and analyzed.HSCs proliferation was detected by MTT method,HSCs cell cycle by flow cytometry,and expression of TGF-β1,Smad 3,Smad 7,α-SMA,collagen I,collgenⅢmRNAs and corresponding proteins by RT-PCR and Western blotting,respectively.【Results】At the normal ALT level,HBV with different viral loads had no significant effect on the proliferation,cell cycle and cell secretion of the HSCs.At the abnormal ALT level,especially when ALT≥2 UNL,with the increase of virus loads,HSCs proliferation accelerated;cells in the G 0/G 1 phase decreased significantly and cells in the S and G 2/M phases increased significantly;the expression of TGFβ1,Smad 3,α-SMA,collagen I,collgenⅢmRNAs and corresponding proteins increased significantly,but Smad 7 mRNA and protein expression decreased significantly,the differences were statistically significant.HBV showed a significantly lower effect on HSCs after the antiviral therapy than before.【Conclusions】This paper reveals the differential effects of HBV on HSCs at different ALT levels and presents a comparative analysis of the effects before and after the antiviral therapy,which provides a theroretical basis for identifying the ALT level as an indication for HBV antiviral therapy.

关 键 词：乙型肝炎病毒 病毒载量 肝星状细胞 细胞周期 细胞增殖 细胞分泌 丙氨酸氨基转移酶 

分 类 号：R512.62[医药卫生—内科学]