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目的了解慢性乙型肝炎患者(CHB)血清中热休克蛋白70(HSP70)与HBV-DNA的关系,并探讨HSP70能否影响乙型肝炎病毒(HBV)的复制与抗原分泌。方法选择安徽医科大学第二附属医院住院和门诊CHB患者60例为研究对象,并以30例健康体检人群为对照者,采用酶联免疫技术(ELISA)分析CHB患者和对照者血清HSP70表达水平,运用荧光定量PCR技术分析CHB患者血清HBV-DNA复制水平;以RNA干扰(RNAi)技术抑制HepG2.2.15细胞中HSP70基因的表达,同时分析细胞培养上清中HBV DNA以及相关抗原的表达情况。结果 CHB患者血清中HSP70的表达水平明显高于其在健康对照者血清中的表达水平(P<0.05),且与HBV-DNA拷贝数呈正相关性(r=0.540,P<0.05);其中HBeAg阳性的CHB患者血清中HSP70的表达水平也显著高于HBeAg阴性CHB患者血清中的表达水平(P<0.05)。以RNAi技术特异性降低HSP70在HepG2.2.15细胞中的表达,HepG2.2.15细胞培养上清中HBV-DNA和相关抗原的分泌量明显降低(P<0.05)。结论 HSP70蛋白很可能在HBV的生命周期中发挥重要作用,从而影响HBV的复制和乙型肝炎的进展。
Objective To investigate the relationship between the expression of heat shock protein 70 (HSP70) and HBV-DNA in patients with chronic hepatitis B (CHB) and to explore whether HSP70 can affect the replication and antigen secretion of hepatitis B virus (HBV). Methods Sixty CHB inpatients and outpatients in the Second Affiliated Hospital of Anhui Medical University were enrolled in this study. Thirty healthy control subjects were used as controls. The serum levels of HSP70 in CHB patients and controls were analyzed by enzyme-linked immunosorbent assay (ELISA) Fluorescence quantitative PCR was used to analyze the level of HBV DNA replication in CHB patients. RNA interference (RNAi) was used to inhibit the expression of HSP70 in HepG2.2.15 cells and the expression of HBV DNA and related antigens in cell culture supernatants. Results The serum level of HSP70 in patients with CHB was significantly higher than that in healthy controls (P <0.05), and positively correlated with HBV-DNA copy number (r = 0.540, P <0.05) The serum levels of HSP70 in patients with positive CHB were also significantly higher than those in patients with HBeAg-negative CHB (P <0.05). The expression of HSP70 in HepG2.2.15 cells was reduced by RNAi technique. The secretion of HBV-DNA and related antigen in HepG2.2.15 cell culture supernatant was significantly decreased (P <0.05). Conclusion HSP70 protein is likely to play an important role in the life cycle of HBV, thus affecting HBV replication and hepatitis B progression.