乙型肝炎病毒与载脂蛋白B关系及其意义的实验研究

Experimental study of relationship between hepatitis B virus and apolipoprotein B and its significance

目的通过检测不同乙型肝炎病毒(HBV)载量受试者血脂水平的差异及其与HBV的关系及其意义。方法选择2017年6月至2018年1月住院的100例HBV携带者及同期入院体检的100例健康人作为研究对象,对其健康状态进行复检,确定受试者无其他疾病影响的条件下,将受试者根据HBV载量分为三组:病毒载量≤500拷贝/ml(正常组,n=100),500拷贝/ml <病毒载量<1×105拷贝/ml(HBV携带组,n=37),病毒载量≥1×105拷贝/ml(乙肝患者组,n=63)。检测血清血脂指标[甘油三脂(TG)、胆固醇(TC)、高密度脂蛋白(HDL)、低密度脂蛋白(LDL)、载脂蛋白(Apo) A1]水平和血浆Apo B水平;对转染与未转染p HBV1. 3质粒的两种Hep G2细胞中Aop B mRNA的表达进行测定;对转染与未转染Apo B的两种Hep G3细胞中HBs Ag、HBe Ag含量及HBV-DNA含量进行测定。结果 HBV数量随着患者病情加重而增多,正常者体内不含HBV。血清学检测显示,随受试者体内病毒载量(正常组→HBV携带组→乙肝患者组)的递增,其血清TC、HDL、LDL、Aop A1和血浆Apo B水平均呈递降(P均<0. 05);其中,LDL含量下降最为明显。进一步对作为HDL主要载脂蛋白的Apo B进行的RT-PCR检测显示,转染p HBV1. 3质粒的Hep G2细胞其Apo B mRNA表达水平低于未转染细胞。HBV指标检测显示,未转染Apo B的Hep G3细胞其HBs Ag、HBe Ag和HBV-DNA含量均明显高于转染细胞(P均<0. 01)。结论 HBV感染可减少患者血脂水平,并以LDL及其主要载脂蛋白Apo B的下降最明显,HBV感染可降低Apo B对HBV复制的抑制作用,促进HBV的复制。该作用是否通过微粒体TC转移蛋白(MTP)途径而产生,有待进一步研究。

Objective By detecting the difference of blood lipid levels in subjects with different hepatitis B virus( BHV)loads to explore its relationship with HBV and their significance. Methods A total of 100 HBV carriers hospitalized from June 2017 to January 2018 and 100 healthy persons for physical examination during the same period were selected as the research subjects. Without the influence of other disease factors determined by re-examined,all subjects were divided into normal group( viral load ≤500 copies/ml,n = 100),HBV carrier group( 500 < viral load < 1 × 10^5 copies/ml,n = 37)and hepatitis B patients group( viral load≥1 × 10^5 copies/ml,n = 63) according to the detection of serum viral load.Serum lipid levels [triglyceride( TG),total cholesterol( TC),high density lipoprotein( HDL),low density lipoprotein( LDL),apolipoprotein( Apo) A1]and plasma Apo B level were measured;the expressions of Apo BmRNA in Hep G2 cells transfected with and without p HBV1. 3 plasmid were determined;the contents of HBs Ag,HBe Ag and HBV-DNA in Hep G3 cells transfected with and without Apo B were detected. Results The loads of HBV increased with the aggravation of the patient’s disease. No hepatitis B virus were detected out in normal peoples. Serological tests showed that the serum levels of TC,HDL,LDL,Aop A1 and plasma level of Apo B decreased gradually with the increase of viral load( normal group→HBV carrier group→hepatitis B patients group,all P < 0. 05),and LDL content decreased most obviously. Further detection of Apo B as the main apolipoprotein of HDL by reverse transcription polymerase chain reaction( RT-PCR) assay showed that Apo B mRNA expression level in Hep G2 cells transfected with p HBV1. 3 plasmid was lower than that in non-transfected cells. Detection of HBV markers showed that the contents of HBs Ag,HBe Ag and HBV DNA in Hep G3 cells without Apo B transfection were significantly higher than those in the Apo B-transfected cells( all P < 0. 01). Conclusions HBV infection can reduce the blood lipid levels,especially LDL and its major apolipoprotein-ApoB. HBV infection can reduce the inhibitory effect of Apo B on HBV replication and promote HBV replication. It needs to be further studied whether this effect is produced through microsomal TC transfer protein pathway.