维生素D及维生素D受体在慢性乙型肝炎患者体内表达水平及抗病毒相关机制的研究

Study on the expression levels and the related antiviral mechanisms of vitamin D and vitamin D receptor in patients with chronic hepatitis B

目的研究维生素D(Vit D)、肝脏维生素D受体(VDR)在慢性乙型肝炎(CHB)患者体内表达水平及其与肝功能、乙型肝炎病毒(HBV)血清学指标相关性,并进一步探讨Vit D体外能否发挥抗病毒作用。方法检测81例初治CHB患者和40例健康体检者血清Vit D含量;采用免疫组化技术分析肝组织VDR表达情况;Vit D单独及联合IFN-α处理HepG2.2.15细胞,检测处理后细胞培养上清液HBsAg、HBeAg、HBV DNA含量,分析各组细胞VDR、IRF-9、MxA蛋白表达水平。结果 CHB组血清Vit D含量低于健康对照(HC)组(P<0.05);CHB组血清Vit D与白蛋白(ALB)呈正相关性(r=0.339,P=0.002),与HBV DNA载量呈负相关性(r=-0.274,P=0.013),与e抗原状态有关(P<0.05),与丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、总胆红素(TBIL)、碱性磷酸酶(ALP)、r-谷氨酰基转移酶(GGT)、HBsAg、HBcAb等无相关性;Vit D、VDR表达与CHB严重程度相关,重度CHB患者血清Vit D、肝组织中VDR表达水平低于轻、中度CHB患者;Vit D+IFN-α联合组较空白组、单独处理组HepG2.2.15细胞VDR、JAK-STAT途径分子IRF-9及下游抗病毒蛋白MxA的表达增强,HBV相关抗原分泌下降。结论慢乙肝患者体内可能存在Vit D/VDR信号通路的功能下降,Vit D可能通过上调VDR表达进而增强IFN-JAK-STAT信号途径发挥协同IFN-α抗病毒效应。

Objective To analyze the expression levels of vitamin D(Vit D),liver vitamin D receptor(VDR)in patients with chronic hepatitis B(CHB)and its correlations with liver function and hepatitis B virus(HBV)serological indicators,and further explore whether Vit D can exert antiviral effects in vitro.Methods The serum levels of Vit D were measured in 81 untreated CHB patients and 40 healthy subjects.The expression of VDR in liver tissue was analyzed by immunohistochemistry.HBV-related parameters(HBsAg,HBeAg,HBV DNA)in cell culture supernatants and the protein expressiones(VDR,IRF-9,MxA)in cells were detected after HepG2.2.15 cellswere treated with Vit D alone or in combination with IFN-α.Results The serum Vit D level in CHB group was lower than that in healthy controls(HC)group(P<0.05).The serum Vit D level was positively correlated with albumia(ALB)(r=0.339,P=0.002),negatively correlated with HBV DNA viral load(r=-0.274,P=0.013),correlated with e atigen status(P<0.05),no correlation with alanine aminotransferase(ALT)、aspartate aminotransferase(AST)、total bilirubin(TBIL)、alkaline phosphatase(ALP)、gamma-glutamyl transferase(GGT),HBsA g and HBc Ab.Also,the expression levels of Vit D and VDR were related to the severity of CHB.The expression levels of Vit D in serum and VDR in liver tissue of severe CHB patients were lower than those of mild and moderate CHB patients.The expression of VDR,JAK-STAT signal transduction molecules IRF-9 and downstream antiviral protein Mx A in the Vit D combination with IFN-αgroup were higher and the secretion of HBV-related antigens was lower than those in the blank group and the single treatment group.Conclusion There may be a decline in the function of the Vit D/VDR signaling pathway in patients with CHB.Vit D may play a synergistic IFN-αantiviral effect by increasing the VDR expression and thereby enhancing the IFN-JAK-STAT signaling pathway.