中国现代医学杂志2024，Vol.34Issue(16)：17-25,9.DOI:10.3969/j.issn.1005-8982.2024.16.004
乙型肝炎病毒通过自分泌运动因子/溶血磷脂酸信号损伤对糖稳态的作用及其机制研究
Hepatitis B virus impairs glucose homeostasis via autotaxin/lysophosphatidic acid signaling
摘要
Abstract
Objective To investigate the effects and underlying mechanisms of hepatitis B virus on impairing glucose homeostasis through the autotaxin(ATX)/lysophosphatidic acid(LPA)signaling.Methods Gene expression databases were used to analyze the effect of HBV on the expression of ATX.Western blotting(WB)was performed to detect the protein expressions of ATX in HBV-expressing cell line HepG2215,HBX-HepG2 and PRES2-HepG2 cells stably expressing HBV trans-regulatory proteins HBx and PreS2,and normal control(NC)HepG2 cells.The dual luciferase reporter assay was applied to detect the effects of HBx and PreS2 on the promoter activity of ATX.HBx-NIT and PreS2-NIT,the mice insulin-secreting cells with stable expressions of HBx and pre-S2,were constructed to detect the effects of HBx and pre-S2 on insulin secretion.Mice models of HBV infection were established by injecting C57BL/6 mice with rAAV8-1.3 HBV via tail veins,while C57BL/6 mice injected with the same volume of normal saline were set as NCs.The glucose tolerance test(GTT)was performed by intraperitoneal injection of glucose at a dosage of 2 g/kg body weight of mice.Serum LPA,serum insulin and blood glucose were measured by liquid chromatography-mass spectrometry,ELISA kits and blood glucose analyzer,respectively.Results The protein expression of ATX in HepG2215 cells was higher than that in HepG2 cells(P＜0.05).The luciferase activity in those co-transfected with PreS2 and ATX promoter was higher than that in those transfected with ATX promoter alone(P＜0.05),and that in those co-transfected with HBX and ATX promoter was also higher compared with that in those transfected with ATX promoter alone(P＜0.05).The protein expression of ATX in HBx-HepG2 cells was higher than that in HepG2 cells(P＜0.05),and that in PreS2-HepG2 cells was higher than that in HepG2 cells(P＜0.05).There were differences in insulin secretion among NIT cells treated with different concentrations of LPA(P＜0.05),and the concentration of LPA was negatively correlated with insulin secretion when it was within the range of 1-3 pmol/L(r=-0.990,P＜0.05).The level of insulin in HBx-NIT cells before treatment with Ki16425 was lower than that in NIT cells(P＜0.05),whereas that in PreS2-NIT cells was lower than that in NIT cells(P＜0.05).The levels of insulin in NIT,HBx-NIT and PreS2-NIT cells after treatment with Ki 16425 were higher than those before the treatment(P＜0.05).The serum level of LPA,the mean concentration of fasting blood glucose,and mean concentrations of blood glucose and the areas under curves(AUCs)of blood glucose concentrations 60 and 120 min after glucose injection in the experimental group were higher than those in the control group(P＜0.05).The serum concentration of insulin and the AUC thereof 15 min after glucose injection in the experimental group were lower than those in the control group(P＜0.05).The receiver operating characteristic(ROC)curve based on blood glucose concentrations in the GTT of HBV-infected mice showed an AUC of 0.770(95％CI:0.556,0.984),a specificity of 60.00％(95％CI:0.122,0.738),and a sensitivity of 90.00％(95％CI:0.555,0.998).The ROC curve based on the concentrations of fasting blood glucose in HBV-infected mice showed an AUC of 0.865(95％CI:0.703,1.027),a specificity of 80.00％(95％CI:0.444,0.975),and a sensitivity of 60.00％(95％CI:0.262,0.878).The homeostasis model assessment-beta cell and the insulin sensitivity index were lower and the insulin resistance index was higher in the experimental group than in the control group(P＜0.05).Conclusions HBV trans-regulatory proteins HBx and PreS2 up-regulate the expression of ATX and enhance the ATX/LPA signaling,leading to inhibition of insulin secretion,abnormal glucose tolerance and impaired glucose homeostasis.关键词
乙型肝炎病毒/反式调节蛋白/自分泌运动因子/溶血磷脂酸信号/糖稳态Key words
hepatitis B virus/trans-regulatory protein/autotaxin/lysophosphatidic acid signaling/glucose homeostasis分类
医药卫生引用本文复制引用
朱政全,梁斌,石明连,谷云艳,周先丽,梁成钦,刘永明,苏何玲..乙型肝炎病毒通过自分泌运动因子/溶血磷脂酸信号损伤对糖稳态的作用及其机制研究[J].中国现代医学杂志,2024,34(16):17-25,9.基金项目
国家自然科学基金(No:81460320,No:82060787,No:31560100) （No:81460320,No:82060787,No:31560100）
