[1]李晓滨,白建云,赵启兵.右美托咪定对心肌缺血再灌注损伤大鼠的作用及机制研究[J].陕西医学杂志,2022,51(9):1060-1065.[doi:DOI:10.3969/j.issn.1000-7377.2022.09.004]
　LI Xiaobin,BAI Jianyun,ZHAO Qibing.Effect and mechanism of dexmedetomidine on myocardial ischemia-reperfusion injury in rats[J].,2022,51(9):1060-1065.[doi:DOI:10.3969/j.issn.1000-7377.2022.09.004]
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右美托咪定对心肌缺血再灌注损伤大鼠的作用及机制研究
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
51
期数:
2022年9期
页码:
1060-1065
栏目:
基础研究
出版日期:
2022-09-05

文章信息/Info

Title:
Effect and mechanism of dexmedetomidine on myocardial ischemia-reperfusion injury in rats
作者:
李晓滨1; 白建云2; 赵启兵3
(1.宝鸡市中医医院麻醉手术科,陕西 宝鸡 721001; 2.榆林市第三医院麻醉科,陕西 榆林 719000; 3.安康市人民医院麻醉科,陕西 安康 725029)
Author(s):
LI Xiaobin; BAI Jianyun; ZHAO Qibing
(Department of Anesthesiology and Surgery,Baoji Hospital of Traditional Chinese Medicine,Baoji 721001,China)
关键词:
心肌缺血再灌注损伤;  右美托咪定;  丝裂原活化蛋白激酶;  氧化应激;  内质网应激;  大鼠
Keywords:
Myocardial ischemia-reperfusion injury;  Dexmedetomidine;  Mitogen-activated protein kinase;  Oxidative stress;  Endoplasmic reticulum stress;  Rat
分类号:
R 542.2
DOI:
DOI:10.3969/j.issn.1000-7377.2022.09.004
文献标志码:
A
摘要:
目的:探讨右美托咪定(Dex)对心肌缺血再灌注损伤(MIRI)大鼠的作用,并分析其机制。方法:将36只雄性SD大鼠随机分为对照组、MIRI模型组(MIRI组)和Dex治疗组(MIRI+Dex组),每组12只。MIRI组和MIRI+Dex组的24只大鼠用于建立MIRI模型。为研究miR-146a在Dex对MIRI大鼠心肌细胞作用中的机制,将心肌细胞分为 NC组(对照组大鼠心肌细胞)、抑制剂组(对照组大鼠心肌细胞转染miR-146a抑制剂)、MIRI+NC组(MIRI组大鼠心肌细胞)、MIRI+抑制剂组(MIRI组大鼠心肌细胞转染miR-146a抑制剂)、Dex+MIRI+NC组(MIRI+Dex组大鼠心肌细胞)和Dex+MIRI+抑制剂组(MIRI+Dex组大鼠心肌细胞转染miR-146a抑制剂)。为进一步验证丝裂原活化蛋白激酶(MAPK)信号通路是否参与了Dex对MIRI心肌细胞的作用机制,将心肌细胞分为对照组(仅用培养基处理大鼠心肌细胞)、SB组[p38 MAPK信号通路抑制剂SB203580(SB)0.5 μmol/L预处理2 h]、MIRI组(MIRI组大鼠心肌细胞)、MIRI+SB组(MIRI组大鼠心肌细胞用0.5 μmol/L SB预处理2 h)、Dex+MIRI组(MIRI+Dex组大鼠心肌细胞)和Dex+MIRI+SB组(MIRI+Dex组大鼠心肌细胞用0.5 μmol/L SB预处理2 h)。使用相应检测试剂盒测定乳酸脱氢酶(LDH)活性、超氧化物歧化酶(SOD)活性和丙二醛(MDA)水平。采用流式细胞术检测细胞凋亡率和活性氧(ROS)水平。使用定量实时PCR(qPCR)检测葡萄糖调节蛋白78(GRP78)、下游靶基因C/EBP家族同源蛋白(CHOP)、锰超氧化物歧化酶(MnSOD)、醌氧化还原酶1(NQO1)、过氧化氢酶(CAT)和miR-146a的mRNA相对表达量。结果:MIRI组miR-146a水平、SOD活性和细胞活力低于对照组,LDH活性、MDA水平、GRP78和CHOP水平、细胞凋亡率高于对照组(均P<0.05)。MIRI+Dex组miR-146a表达水平、SOD活性和细胞活力高于MIRI组,LDH活性、MDA水平、GRP78和CHOP水平、细胞凋亡率低于MIRI组(均P<0.05)。抑制剂组和MIRI+NC组细胞活力和miR-146a mRNA相对表达量明显低于NC组,Dex+MIRI+NC组细胞活力和miR-146a mRNA相对表达量高于Dex+MIRI+抑制剂组,而ROS水平表现出相反趋势(均P<0.05)。抑制剂组和MIRI+NC组GRP78、CHOP mRNA相对表达量高于NC组,MIRI+NC组低于MIRI+抑制剂组而高于Dex+MIRI+NC组,且Dex+MIRI+抑制剂组高于Dex+MIRI+NC组,而CAT、MnSOD、NQO1 mRNA相对表达量显示出相反趋势(均P<0.05)。MIRI组细胞活力低于对照组,但在MIRI+SB组和Dex+MIRI组中升高(均P<0.05)。MIRI组GRP78、CHOP mRNA相对表达量高于对照组,MIRI+SB和Dex+MIRI组则低于MIRI组(均P<0.05)。但Dex+MIRI+SB组GRP78 mRNA相对表达量低于Dex+MIRI组,CHOP mRNA相对表达量高于Dex+MIRI组(均P<0.05)。经SB预处理2 h后发现,抑制剂组细胞活力低于NC组,而抑制剂+SB组细胞活力高于抑制剂组(均P<0.05); 抑制剂组GRP78、CHOP mRNA相对表达量高于NC组和抑制剂+SB组(均P<0.05); 抑制剂+SB组GRP78、CHOP mRNA相对表达量低于抑制剂(均P<0.05)。结论:Dex可能通过调节miR-146a的表达,进一步调节内质网应激(ERS)和氧化应激,最终通过MAPK信号通路影响MIRI心肌细胞的细胞活力和凋亡。
Abstract:
Objective:To investigate the effect of dexmedetomidine(Dex)on myocardial ischemia-reperfusion injury(MIRI)in rats and analyze its mechanism.Methods:A total of 36 male SD rats were randomly divided into control group,MIRI model group(MIRI group)and Dex treatment group(MIRI+Dex group),with 12 rats in each group.Twenty-four rats in MIRI group and MIRI+Dex group were used to establish the MIRI model.In order to study the mechanism of miR-146a in the action of Dex on MIRI rat cardiomyocytes,cardiomyocytes were divided into NC group(rat cardiomyocytes in control group),inhibitor group(rat cardiomyocytes in control group were transfected with miR-146a inhibitor),MIRI+NC group(rat cardiomyocytes in MIRI group),MIRI+inhibitor group(rat cardiomyocytes in MIRI group were transfected with miR-146a inhibitor),Dex+MIRI+NC group(rat cardiomyocytes in MIRI+Dex group)and Dex+MIRI+inhibitor group(rat cardiomyocytes in MIRI+Dex group were transfected with miR-146a inhibitor).In order to further verify whether the mitogen-activated protein kinase(MAPK)signaling pathway was involved in the mechanism of Dex on MIRI cardiomyocytes,cardiomyocytes were divided into control group(rat cardiomyocytes was only treated with culture medium),SB group [rat cardiomyocytes were pretreated with 0.5 μmol/L p38 MAPK signaling pathway inhibitor SB203580(SB)for 2 hours],MIRI group(rat cardiomyocytes in MIRI group),MIRI+SB group(rat cardiomyocytes in MIRI group were pretreated with 0.5 μmol/L SB for 2 hours),Dex+MIRI group(rat cardiomyocytes in MIRI+Dex group)and Dex+MIRI+SB group(rat cardiomyocytes in MIRI+Dex group were pretreated with 0.5 μmol/L SB for 2 hours).Lactate dehydrogenase(LDH)activity,superoxide dismutase(SOD)activity,and malondialdehyde(MDA)levels were determined by using corresponding detection kits.Flow cytometry was performed to determine apoptosis rate and reactive oxygen species(ROS)level.Quantitative real-time PCR(qPCR)was used to detect the relative mRNA expression of GRP78,CHOP,MnSOD,NQO1,CAT and miR-146a.Results:MiR-146a level,SOD activity and cell viability in MIRI group were lower than those in control group,while LDH activity,MDA,GRP78 and CHOP levels,and apoptosis rate were higher than those in control group(all P<0.05).MiR-146a level,SOD activity and cell viability in MIRI+Dex group were higher than those in MIRI group,while LDH activity,MDA,GRP78 and CHOP levels,and apoptosis rate were lower than those in MIRI group(all P<0.05).The cell viability and relative expression of miR-146a mRNA in inhibitor group and MIRI+NC group were significantly lower than those in NC group,and the cell viability and relative expression of miR-146a mRNA in Dex+MIRI+NC group were higher than those in Dex+MIRI+inhibitor group,and the ROS level showed the opposite trend(all P<0.05).The relative mRNA expression levels of GRP78 and CHOP in inhibitor group and MIRI+NC group were higher than those in NC group,and those in MIRI+NC group were lower than those in MIRI+inhibitor group but higher than those in Dex+MIRI+NC group,and those in Dex+MIRI+inhibitor group were higher than those in Dex+MIRI+NC group,while the relative mRNA expression levels of CAT,MnSOD and NQO1 showed opposite trend(all P<0.05).Cell viability in MIRI group was lower than that in control group,but increased in MIRI+SB group and Dex+MIRI group(all P<0.05).The relative mRNA expression levels of GRP78 and CHOP in MIRI group were higher than those in control group,while those in MIRI+SB group and Dex+MIRI group were lower than those in MIRI group(all P<0.05).However,the relative mRNA expression level of GRP78 in Dex+MIRI+SB group was lower than that in Dex+MIRI group,and the relative mRNA expression level of CHOP was higher than that in Dex+MIRI group(all P<0.05).After pretreatment with SB for 2 hours,it was found that the cell viability of inhibitor group was lower than that of NC group,while that of inhibitor+SB group was higher than that of inhibitor group(all P<0.05).The relative mRNA expression levels of GRP78 and CHOP in inhibitor group were higher than those in NC group and inhibitor+SB group(all P<0.05).Conclusion:Dex may further regulate endoplasmic reticulum stress and oxidative stress by regulating the expression of miR-146a,and finally affect the cell viability and apoptosis of MIRI cardiomyocytes through the MAPK signaling pathway.

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备注/Memo

备注/Memo:
基金项目:陕西省榆林市科协青年人才托举计划项目(20190109)
更新日期/Last Update: 2022-09-05