摘要
目的探讨混合谱系激酶结构域样蛋白(MLKL)、caspase-8及受体相互作用蛋白激酶3(RIPK3)mRNA在不同阶段乙型肝炎病毒(HBV)感染患者中的表达水平及诊断价值。方法选择2016年11月至2019年11月新乡市传染病医院收治的175例HBV感染患者为研究对象,根据病情程度将患者分为慢性乙型肝炎(CHB)组(n=67)、肝硬化(LC)组(n=61)和肝细胞肝癌(HCC)组(n=47);另选择同期于本院体检的80例体检健康者为对照组。采集各组受试者外周静脉血,应用实时荧光定量聚合酶链反应法检测外周血单个核细胞(PBMC)中MLKL、caspase-8及RIPK3 mRNA表达水平;采用Pearson相关检验分析HBV感染不同阶段患者PBMC中MLKL、caspase-8及RIPK3 mRNA表达水平的相关性,受试者操作特征曲线下面积(AUC)评估MLKL、caspase-8及RIPK3 mRNA表达水平对不同阶段HBV感染患者的鉴别诊断价值。结果4组受试者PBMC中MLKL、RIPK3、caspase-8 mRNA表达水平比较差异有统计学意义(F=1019.364、1009.381、159.407,P<0.05)。CHB组、LC组、HCC组患者PBMC中MLKL、RIPK3 mRNA表达水平显著高于对照组,caspase-8 mRNA表达水平显著低于对照组(P<0.05)。LC组、HCC组患者PBMC中MLKL、RIPK3 mRNA表达水平显著高于CHB组,caspase-8 mRNA表达水平显著低于CHB组(P<0.05)。HCC组患者PBMC中MLKL、RIPK3 mRNA表达水平显著高于LC组,caspase-8 mRNA表达水平显著低于LC组(P<0.05)。Pearson相关检验结果显示,不同阶段HBV感染组患者PBMC中MLKL mRNA表达与RIPK3 mRNA表达呈正相关(r=0.414、0.432、0.449,P<0.01),MLKL mRNA表达与caspase-8 mRNA表达呈负相关(r=-0.556、-0.378、-0.721,P<0.01),RIPK3 mRNA表达与caspase-8 mRNA表达呈负相关(r=-0.415、-0.400、-0.416,P<0.01)。PBMC中MLKL、caspase-8及RIPK3 mRNA表达水平鉴别CHB和HCC的AUC分别为0.918、0.859和0.912,三者联合鉴别CHB和HCC的AUC为0.945。PBMC中MLKL、caspase-8及RIPK3 mRNA表达水平鉴别LC和HCC的AUC分别为0.768、0.834和0.839,三者联合鉴别LC和HCC的AUC为0.895。结论HBV感染不同阶段患者PBMC中MLKL、caspase-8及RIPK3 mRNA表达水平显著上升,且三者之间存在显著相关性,MLKL、caspase-8及RIPK3 mRNA表达水平可作为鉴别诊断不同阶段HBV感染患者的生物学标志物。
Objective To investigate the expression and diagnostic value of mixed lineage kinase domain-like protein(MLKL),caspase-8 and receptor interacting protein kinase 3(RIPK3)mRNA in patients at different stages of hepatitis B virus(HBV)infection.Methods A total of 175 HBV infected patients admitted to Xinxiang Infectious Disease Hospital from November 2016 to November 2019 were selected as the study subjects,and they were divided into chronic hepatitis B(CHB)group(n=67),liver cirrhosis(LC)group(n=61)and hepatocellular carcinoma(HCC)group(n=47)according to the disease severity of patients.Another 80 healthy subjects who underwent physical examination in the hospital during the same period were selected as the control group.Peripheral venous blood was collected from all subjects and the expression levels of MLKL,caspase-8 and RIPK3 mRNA in peripheral blood mononuclear cell(PBMC)were detected by quantitative real-time fluorescence polymerase chain reaction.The correlation between the MLKL,caspase-8 and RIPK3 mRNA expression levels in PBMC of patients with HBV infection at different stages was analyzed by Pearson correlation test.The differential diagnostic value of expression levels of MLKL,caspase-8 and RIPK3 mRNA on different stages of HBV infection was evaluated by area under the curve(AUC)in receiver operating characteristic curve.Results There was significant difference in the expression levels of MLKL,RIPK3 and caspase-8 mRNA in PBMC of subjects among the four groups(F=1019.364,1009.381,159.407;P<0.05).The expression levels of MLKL and RIPK3 mRNA in PBMC of patients in the CHB,LC and HCC groups were significantly higher than those in the control group,while the expression level of caspase-8 mRNA was significantly lower than that in the control group(P<0.05).The expression levels of MLKL and RIPK3 mRNA in PBMC of patients in the LC and HCC groups were significantly higher than those in the CHB group,while the expression level of caspase-8 mRNA was significantly lower than that in the CHB group(P<0.05).The expression levels of MLKL and RIPK3 mRNA in PBMC of patients in the HCC group were significantly higher than those in the LC group,while the expression level of caspase-8 mRNA was significantly lower than that in the LC group(P<0.05).Pearson correlation test results showed that the expression of MLKL mRNA was positively correlated with the expression RIPK3 mRNA in PBMC of patients with different stages of HBV infection(r=0.414,0.432,0.449;P<0.01);the expression of MLKL mRNA was negatively correlated with the expression of caspase-8 mRNA(r=-0.556,-0.378,-0.721;P<0.01);and the expression of RIPK3 mRNA was negatively correlated with the expression of caspase-8 mRNA(r=-0.415,-0.400,-0.416;P<0.01).The AUC of MLKL,caspase-8 and RIPK3 mRNA expression levels in PBMC in identifying of CHB and HCC was 0.918,0.859,0.912,respectively;the AUC of above three indexes in combination in identifying of CHB and HCC was 0.945.The AUC of MLKL,caspase-8 and RIPK3 mRNA expression levels in PBMC in identifying of LC and HCC was 0.768,0.834 and 0.839,respectively;the AUC of above three indexes in combination in identifying of LC and HCC was 0.895.Conclusion The expression levels of MLKL,caspase-8 and RIPK3 mRNA in PBMC of patients at different stages of HBV infection are significantly increased;and there is a significant correlation among the three indicators.The expression levels of MLKL,caspase-8 and RIPK3 mRNA can be used as biological markers for differential diagnosis of patients at different stages of HBV infection.
作者
文正燕
孙传俊
杨献光
WEN Zhengyan;SUN Chuanjun;YANG Xianguang(Department of Hepatology,Xinxiang Infectious Disease Hospital,Xinxiang 453000,Henan Province,China;Department of Clinical Laboratory,Xinxiang Infectious Disease Hospital,Xinxiang 453000,Henan Province,China;School of Life Science,Henan Normal University,Xinxiang 453000,Henan Province,China)
出处
《新乡医学院学报》
CAS
2023年第7期628-633,共6页
Journal of Xinxiang Medical University
基金
河南省重点科技攻关项目(编号:222102310244)。
