中介素对缺氧、缺糖诱导小鼠HL-1细胞焦亡的作用  

Effect of intermedin on pyroptosis of murine HL-1 cells induced by hypoxia and glucose deprivation

作　　者：李礼 刘尊国 杨生平[1] 孙红茜[3] 薛莉[1] LI Li;LIU Zunguo;YANG Shengping;SUN Hongqian;XUE Li(Department of Cardiovascular Medicine,General Hospital of Ningxia Medical University,Yinchuan 750004,China;Department of Geriatrics and Specialty Medicine,General Hospital of Ningxia Medical University,Yinchuan 750004,China;ECG Room,General Hospital of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学总医院心血管内科,宁夏银川750004 [2]宁夏医科大学总医院老年与特需医学科,宁夏银川750004 [3]宁夏医科大学总医院心电图室,宁夏银川750004

出　　处：《宁夏医学杂志》2025年第1期19-23,共5页Ningxia Medical Journal

基　　金：宁夏自然科学基金项目(2022AAC03480);宁夏医科大学校级项目(XM2021008)。

摘　　要：目的研究中介素(IMD)对小鼠心肌细胞(HL-1)缺氧、缺糖损伤后细胞焦亡的影响及分子机制。方法将体外培养的HL-1细胞分为对照组(Control组)、急性心肌梗死模拟组(AMI组)、急性心肌梗死模拟+IMD干预组(AMI+IMD组),各组细胞分别处理0 h、4 h、12 h、24 h。在光镜下观察各组细胞形态;采用Western blot方法检测各组细胞天冬氨酸蛋白水解酶1(Caspase-1)、天冬氨酸蛋白水解酶11(Caspase-11)、核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)、消化素家族D蛋白分子(GSDMD)焦亡相关蛋白表达水平;采用荧光定量PCR法验证各组细胞Caspase-1、Caspase-11、NLRP3、GSDMD的mRNA表达水平。结果与Control组比较,AMI组细胞形态在显微镜下表现为细胞体积肿胀膨大、破裂及细胞膜起泡等细胞焦亡典型改变,在24 h时焦亡损伤最严重,而经过IMD干预可以缓解这种现象;与Control组比较,AMI组细胞在4 h、12 h、24 h时Caspase-1、Caspase-11、NLRP3、GSDMD蛋白表达均明显升高(P<0.05);与AMI组比较,AMI+IMD组细胞在4 h、12 h、24 h时Caspase-1、Caspase-11、NLRP3、GSDMD蛋白表达均明显下调(P<0.05)。与Control组比较,AMI组细胞在4 h、12 h、24 h时Caspase-1、Caspase-11、NLRP3、GSDMD的mRNA表达均显著上升(P<0.05);与AMI组比较,AMI+IMD组细胞在4 h时仅Caspase-1的mRNA表达水平开始有所下调,在12 h时Caspase-1、Caspase-11、NLRP3、GSDMD的mRNA表达水平均出现显著下调,在24 h时Caspase-1、Caspase-11、NLRP3的mRNA表达水平继续下调(P<0.05),而GSDMD的mRNA表达水平与AMI组比较,差异无统计学意义(P>0.05)。结论IMD可减轻缺氧、缺糖诱导的HL-1细胞焦亡,改善急性心肌梗死的发展,此作用可能与抑制NLRP3/Caspase-1/GSDMD焦亡经典通路和Caspase-11/GSDMD非经典通路有关。Objective To investigate the effect and molecular mechanism of intermedin(IMD)on pyroptosis of mice cardiomyocytes(HL-1)after hypoxia and glucose deprivation injury.Methods HL-1 cells in vitro culture divided into control group(Control group),acute myocardial infarction simulation group(AMI group),and acute myocardial infarction simulation+IMD intervention group(AMI+IMD group),and the cell of each group processed separately 0 h,4 h,12 h and 24 h.The morphology of cells in each group was observed under light microscopy.Western-blot method was used to detect the expression levels of proptosis-related proteins of Caspase-1,Caspase-11,NLRP3 and Gasdermin D(GSDMD)in each group.The mRNA expression levels of Caspase-1,Caspase-11,NLRP3 and GSDMD were verified by quantitative PCR.Results Compared with the control group,the morphology of cells in the AMI group showed typical changes of pyroptosis,such as cell volume swelling,rupture,and cell membrane blistering,and the pyroptosis damage was the most serious at 24 h,which could be alleviated by IMD intervention.Compared with the control group,the protein expressions of Caspase-1,Caspase-11,NLRP3 and GSDMD were significantly increased at 4 h,12 h and 24 h in the AMI group(P<0.05),and compared with the AMI group,the proteins of Caspase-1,Caspase-11,NLRP3 and GSDMD in the AMI+IMD group were significantly down-regulated at 4h,12h and 24h(P<0.05).Compared with the control group,the mRNA expressions of Caspase-1,Caspase-11,NLRP3 and GSDMD in the AMI group were significantly increased at 4 h,12 h and 24 h(P<0.05);Compared with the AMI group,only the mRNA expression level of Caspase-1 in the AMI+IMD group began to be down-regulated at 4h,the mRNA expression levels of Caspase-1,Caspase-11,NLRP3 and GSDMD were significantly down-regulated at 12h,and the mRNA expression levels of Caspase-1,Caspase-11 and NLRP3 continued to be down-regulated at 24 h(P<0.05),while the mRNA expression level of GSDMD was not statistically significant compared with that of the AMI group(P>0.05).Conclusion

关 键 词：中介素 心肌细胞 细胞焦亡 消化素家族D蛋白分子 

分 类 号：R541.4[医药卫生—心血管疾病]