TXNIP、NLRP3在冠状动脉粥样硬化斑块中的表达情况及其与冠心病猝死的关系

Expression of TXNIP,NLRP3 in coronary atherosclerotic plaque and their relationship with sudden death of coronary heart disease

目的探讨TXNIP、NLRP3在冠状动脉粥样硬化斑块中的表达及其与斑块继发病变、冠心病猝死的关系。方法回顾性分析2019年1月至2022年3月贵州医科大学法医司法鉴定中心经尸体解剖提取的105例心脏冠状动脉标本及来源者相关资料。根据冠状动脉有无硬化斑块将其分为无病变组(n=20)和斑块组(n=85),再根据冠状动脉是否存在继发病变及其是否为冠心病猝死案例将斑块组分为非冠心病猝死组(n=25)、冠心病猝死无继发病变组(n=30)和冠心病猝死合并继发病变组(n=30)。苏木素-伊红(HE)染色制片及利用IPP6.0图像分析软件测量冠状动脉内膜及病灶厚度、纤维帽厚度、坏死灶厚度及管腔狭窄程度。免疫组织化学法、Western blot和实时荧光定量逆转录-PCR(qRT-PCR)检测冠状动脉TXNIP、NLRP3分布特点及表达情况。结果与无病变组比较,其余3组内膜及病灶厚度、纤维帽厚度、坏死灶厚度更厚,管腔狭窄程度更高,差异有统计学意义(P<0.05)。与冠心病猝死无继发病变组比较,冠心病猝死合并继发病变组内膜及病灶厚度、坏死灶厚度更厚,管腔狭窄程度更高,差异有统计学意义(P<0.05)。无病变组冠状动脉管壁未见TXNIP、NLRP3蛋白表达。非冠心病猝死组TXNIP、NLRP3蛋白强阳性表达率为40.0%、36.0%,弱阳性表达率为32.0%、36.0%,较弱阳性表达率为28.0%、28.0%;冠心病猝死无继发病变组强阳性表达率为50.0%、43.3%,较强阳性表达率为33.3%、36.7%,弱阳性表达率为16.7%、20.0%;冠心病猝死合并继发病变组强阳性表达率为73.3%、76.7%,较强阳性表达率为26.7%、23.3%。冠心病猝死合并继发病变组冠状动脉TXNIP、NLRP3蛋白、mRNA表达水平高于其余3组,差异有统计学意义(P<0.05)。冠状动脉斑块内TXNIP与NLRP3表达的吸光度值、蛋白及mRNA水平呈正相关(P<0.05)。TXNIP、NLRP3表达水平与内膜及病灶厚度呈正相关,与纤维帽厚度呈负相关(P<0.05);冠状动脉坏死灶厚度与TXNIP、NLRP3呈正相关(P<0.05)。结论TXNIP、NLRP3可作为冠心病猝死的诊断指标。

Objective To investigate the expression of TXNIP and NLRP3 in atherosclerotic plaque of coronary artery and their relationship with secondary lesion of plaque and sudden death of coronary heart disease.Methods A total of 105 cases of cardiac coronary samples extracted from autopsy anatomy and related data in the Forensic Judicial Appraisal Center of Guizhou Medical University from January 2019 to March 2022 were analyzed retrospectively.They were divided into the non-lesion group(n=20)and plaque group(n=85)according to whether or not having harden plaque in coronary artery.Then the plaque group was divided into secondary lesion group(n=30)and coronary heart disease sudden death complicating secondary lesion group(n=30).The hematoxylin-eosin(HE)dyed section was prepared.The IPP6.0 image analysis software was used to measure the thickness of coronary intima and lesion,the thickness of fibrous cap,the thickness of necrotic lesion and the degree of lumen stenosis.Immunohistochemical method,Western blot and real-time fluorescent quantitative reverse transcription-PCR(qRT-PCR)were used to detect the distribution characteristics and expression levels of TXNIP and NLRP3 in coronary arteries.Results Compared with the non-lesion group,the thickness of the intima,lesion,fibrous cap and necrosis lesion in the other three groups was thicker,the stenosis degree of lumen was higher,and the differences were statistically significant(P<0.05).Compared with the coronary heart sudden death without secondary lesion group,the thickness of the intima,lesion and necrose lesion in the coronary heart disease sudden death complicating secondary lesion group was thicker,the necrosis degree of lumen was higher,and the differences were statistically significant(P<0.05).The TXNIP and NLRP3 proteins expressions were not seen in the coronary arterial wall of the no-lesion group.The strong positive expression rates of TXNIP and NLRP3 in the non-coronary sudden death group were 40.0%and 36.0%,the weak positive expression rates were 32.0%and 36.0%,and the weaker positive expression rates were 28.0%and 28.0%.The strong positive expression rates in the coronary heart disease sudden death without secondary lesion group were 50.0%and 43.3%,the stronger positive expression rates were 33.3%and 36.7%,and the weak positive expression rates were 16.7%and 20.0%;the strong positive expression rates in the coronary heart disease sudden death complicating secondary lesion group were 73.3%and 76.7%,the stronger positive expression rates were 26.7%and 23.3%.The coronary artery TXNIP and NLRP protein and mRNA levels in the coronary heart disease sudden death complicating secondary lesion group were higher than those in the other three groups with statistical difference(P<0.05).TXNIP in coronary arterial plaque was positively correlated with the absorbance value of NLRP3 expression absorbance value,protein and mRNA expression level(P<0.05).The TXNIP and NLRP3 expression levels were positively correlated with the intima and lesion thickness,and negatively correlated with the fibrous cap thickness(P<0.05).The necrosis lesion area of coronary artery was positively correlated with the TXNIP and NLRP3(P<0.05).Conclusion TXNIP and NLRP3 could serve as the diagnostic indicators of coronary heart disease sudden death.