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| 地氟醚通过上调miR-29b表达减轻大鼠的心肌损伤 |
| Desflurane alleviates myocardial injury of rats by up-regulating miR-29b expression |
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| DOI：10.12089/jca.2021.11.014 |
| 中文关键词: 地氟醚 心肌梗死 微小RNA-29b 信号转导和转录激活因子3 心肌保护 |
| 英文关键词: Desflurane Myocardial infarction MiR-29b Signal transducer and activator of transcription 3 Myocardial protection |
| 基金项目:武汉市卫生计生委科研计划资助项目（WX20D40） |
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| 中文摘要: |
目的 探讨微小RNA-29b（miR-29b）在地氟醚处理减轻大鼠心肌梗死中的作用。
方法 成年雄性健康Wistar大鼠80只，10周龄，体重350～400 g，随机分为五组：假手术组（S组）、心肌梗死组（MI组）、地氟醚组（MD组）、空载AAV9组（MA组）和miR-29b沉默组（MR组），每组16只。S组仅分离冠状动脉不结扎，其余组均建立心肌梗死模型，S组和MI组造模前3 d经尾静脉注射磷酸缓冲液（PBS），MD组造模前3 d经尾静脉注射PBS，并于造模前30 min吸入5.9%地氟醚1 h，之后吸入1.3%地氟醚维持麻醉，MA组和MR组造模前3 d分别经尾静脉注射AAV9、AAV9-miR-29b-sponge，余同MD组。再灌注120 min后，采用Werstern blot法检测心肌组织转录激活因子3（STAT3）、p-STAT3、白介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）蛋白含量；RT-qPCR检测心肌组织miR-29b表达量；红四氮唑（TTC）和HE染色观察心肌梗死面积百分比和组织形态；TargetScan预测并用双荧光素酶实验验证miR-29b与STAT3的靶向关系。
结果 与S组比较，MI组STAT3、p-STAT3、IL-1β、TNF-α蛋白相对含量明显升高，miR-29b相对表达量明显降低，心肌梗死面积百分比明显增大（P＜0.05），心肌组织可见炎症浸润等损伤。与MI组比较，MD组STAT3、p-STAT3、IL-1β、TNF-α蛋白相对含量明显降低，miR-29b相对表达量明显升高，心肌梗死面积百分比明显减小（P＜0.05），心肌组织损伤减轻。与MA组比较，MR组STAT3、p-STAT3、IL-1β、TNF-α蛋白相对含量明显升高，miR-29b相对表达量明显降低，心肌梗死面积百分比明显增大（P＜0.05），心肌组织损伤加重。转染STAT3-3′UTR-WT、miR-29b mimics的心肌细胞荧光素酶活性明显弱于转染STAT3-3′UTR-WT、miR-29b NC的心肌细胞（P＜0.05）。
结论 地氟醚处理可通过上调miR-29b表达，靶向抑制STAT3表达，抑制STAT3相关炎症反应，减轻大鼠心肌梗死损伤，发挥心肌保护作用。 |
| 英文摘要: |
Objective To investigate the role of microRNA-29b (miR-29b) in reducing myocardial infarction in rats treated by desflurane.
Methods Eighty Wistar rats, aged 10 weeks and weighing 350-400 g, were randomly divided into five groups: sham operation group (group S), myocardial infarction group (group MI), desflurane group (group MD), empty AAV9 group (group MA) and miR-29b silence group (group MR), 16 rats in each group. Only the rats of group S were isolated their coronary artery without ligation, the rats of other group established the model of myocardial infarction. The rats of group S and group MI were injected PBS via tail vein 3 days before modeling. The rats of group MD were injected PBS via tail vein 3 days before modeling, and inhaled 5.9% desflurane 30 minutes before modeling, and inhaled 1.3% desflurane to maintain anesthesia. The rats of group MA and group MR were respectively injected AAV9 and AAV9-miR-29b-sponge via tail vein 3 days before modeling, and both inhaled 5.9% desflurane 30 minutes before modeling, and inhaled 1.3% desflurane to maintain anesthesia. 120 minutes after reperfusion, Western blot was used to detect the protein expression of signal transducer and activator of transcription 3(STAT3), p-STAT3, interleukin(IL-1β), tumor necrosis factor(TNF)-α; RT-qPCR was used to detect the miR-29b expression; 2,3,5-triphenyl-2H-tetrazolium chloride(TTC) staining and hematoxylin-eosin(HE) staining were performed to observe the myocardial infarction volume and histological morphology, TargetScan was used to predict the targeting relationship between miR-29b and STAT3 and dual luciferase experiment was used to verify it.
Results Compared with group S, STAT3, p-STAT3, IL-1β, TNF-α protein levels and myocardial infarction area percentage increased in group MI, and miR-29b level decreased (P < 0.05), the myocardial tissue showed inflammation infiltration. Compared with group MI, STAT3, p-STAT3, IL-1β, TNF-α protein levels and myocardial infarction area percentage decreased in group MD, and miR-29b level increased (P < 0.05), the myocardial tissue damage was reduced. Compared with group MA, STAT3, p-STAT3, IL-1β, TNF-α protein levels and myocardial infarction area percentage increased in group MR, and miR-29b level decreased (P < 0.05), the myocardial tissue damage was aggravated. The luciferase activity of cardiomyocytes transfect of STAT3-3′UTR-WT and miR-29b mimics group was lower than that of STAT3-3′UTR-WT and miR-29b NC group (P < 0.05).
Conclusion Desflurane treatment may up-regulate the expression of miR-29b, targeting inhibition of STAT3 expression, inhibiting STAT3-related inflammation, thus reduces injury and protects myocardial tissue in myocardial infarction rats. |
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