The role of miRNA-1 in cardiomyocyte pyroptosis and injury after cardiac arrest cardiopulmonary resuscitation in rats
YIN Jiana1, LI Bingcan1, ZHOU Peisen2, LEI Yuanli1, WANG Dongsheng1, XU Huaqing1, SONG Wenxing1, HE Aiwen1, LI Zhangping3
1.Department of Emergency, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325015, China; 2.Department of Emergency, the Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325027, China; 3.Department of Emergency, the Quzhou Hospital Affiliated to Wenzhou Medical University, Quzhou People’s Hospital, Quzhou 324000, China
YIN Jiana,LI Bingcan,ZHOU Peisen, et al. The role of miRNA-1 in cardiomyocyte pyroptosis and injury after cardiac arrest cardiopulmonary resuscitation in rats[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2023, 53(1): 29-35，41.
摘要目的：探讨微小RNA-1（miR-1）在大鼠心搏骤停/心肺复苏（CA/CPR）后心肌细胞焦亡与损伤中的作用。方法：心搏骤停/心肺复苏模型的建立和分组：SD大鼠按照随机数字表法分为心搏骤停复苏（CA）组和假手术对照（Sham）组。前者再分为5组：CA组，CA+antago miR-1组，CA+antago miR NC组，CA+ago miR-1组，CA+ago miR NC组，每组n=6。所有6组大鼠在自主循环恢复（ROSC）后6 h取心尖部心肌组织、血样待测。用Real-time PCR法检测各组心肌细胞中miR-1的mRNA水平，Western blot法检测各组心肌细胞中焦亡相关的蛋白NLRP3、Caspase-1、IL-1β、IL-18的表达情况，用TUNEL染色技术检测心肌细胞焦亡率，ELISA技术检测血清中CK-MB、cTnI浓度。结果：各组大鼠体质量、基础平均动脉压（MAP）差异无统计学意义（P＞0.05）。与Sham组比较，CA组大鼠miR-1表达水平、心肌焦亡相关蛋白NLRP3、Caspase-1、IL-1β、IL-18表达、焦亡率、血清CK-MB与cTnI浓度升高（P＜0.05）。与CA组比，CA+antago miR-1组大鼠miR-1表达水平、心肌焦亡相关蛋白NLRP3、Caspase-1、IL-1β、IL-18表达、焦亡率、血清CK-MB与cTnI浓度降低（P＜0.05），CA+ago miR-1组大鼠miR-1表达水平、心肌焦亡相关蛋白NLRP3、Caspase-1、IL-1β、IL-18表达、焦亡率、血清CK-MB与cTnI浓度升高（P＜0.05），CA+antago miR NC组与CA+ago miR NC组大鼠miR-1表达水平、心肌焦亡相关蛋白NLRP3、Caspase-1、IL-1β、IL-18表达、焦亡率、血清CK-MB与cTnI浓度差异无统计学意义（P＞0.05）。结论：CA/CPR大鼠ROSC后存在明显心肌焦亡和心肌损伤，静脉使用ago miR-1和antago miR-1可以调节CA/CPR后大鼠心肌miR-1的表达，miR-1促进大鼠CA/CPR后心肌焦亡和心肌损伤，miR-1调节大鼠CA/CPR后心肌损伤机制可能和其促进心肌细胞焦亡相关。
Abstract：Objective: To investigate the role of miR-1 in myocardial pyroptosis and injury after cardiac arrest/cardiopulmonary resuscitation (CA/CPR) in rats. Methods: Cardiac arrest/cardiopulmonary resuscitation model was establishment and Spragu-Dawley (SD) rats were divided as cardiac arrest resuscitation (CA) group and sham operation control (Sham) group according to random number table method. The former was subdivided into 5 groups: CA group, CA+antago miR-1 group, CA+antago miR NC group, CA+ago miR-1 group, CA+ago miR NC group (n=6). All cardiac apical myocardial tissue and blood samples of all 6 groups were tested 6 hours after ROSC. The mRNA level of miRNA-1 in each group was detected using Real-time PCR. The expression of pyroptosis-related proteins including NLRP3, Caspase-1, IL-1β and IL-18 in each group was detected by Western blot. The pyroptosis rate of cardiomyocytes was detected by Tunel stain assay.The concentration of CK-MB and cTnI in serum was detected by ELISA. Results: There were no significant difference in body weight and basic MAP among the groups (P>0.05). Compared with Sham group, CA group had an increased expression of miR-1, NLRP3, Caspase-1, IL-1β, IL-18, pyroptosis rate, serum CK-MB and cTnI concentration (P<0.05). Compared with CA group, CA+antago miR-1 group had decreased expression of miR-1, NLRP3, Caspase-1, IL-1β, IL-18, pyroptosis rate, serum CK-MB and cTnI concentration (P<0.05); the expression of miR-1 and the expression of myocardial pyroptosis related proteins NLRP3, Caspase-1, IL-1β, IL-18, pyroptosis rate, serum CK-MB and cTnI concentration increased in CA+ago miR-1 group (P<0.05); there was no significant change in the expression of miR-1, NLRP3, Caspase-1, IL-1β, IL-18, pyroptosis rate, serum CK-MB and cTnI concentration in CA+antago miR NC group and CA+ago miR NC group (P>0.05). Conclusion: There was obvious myocardial pyroptosis and injury in CA/CPR rats after ROSC. Intravenous administration of ago miR-1 and antago miR-1 could regulate the expression of miR-1 in rat cardiomyocytes after CA/CPR, and miR-1 promoted myocardial pyroptosis and injury in rats after CA/CPR. The mechanism of miR-1 regulating myocardial injury after CA/CPR might be related to its promotion of myocardial pyroptosis.
