摘要
目的探讨miR-21对小鼠心脏缺血再灌注损伤的影响及其机制。方法 40只C57BL/6J小鼠随机分为对照组、模型组、agomiR-21组和antagomiR-21组,结扎心冠状动脉前降支进行缺血再灌注损伤造模。agomiR-21组和antagomiR-21组分别给予miR-21的激动剂和拮抗剂,对照组和模型组给予等体积的生理盐水。给药4周后,Elisa检测血清炎症因子TNF-α、IL-6、IL-1β表达水平;HE染色检测心肌组织病理变化,原位缺口末端转移酶标记法(TUNEL)检测小鼠心肌细胞凋亡水平;Western blot检测各组小鼠心肌组织中程序性细胞死亡因子4(PDCD4)的蛋白表达量;荧光素酶报告实验验证miR-21与PDCD4靶向关系;心肌细胞H9C2体外转染PDCD4的siRNA,然后流式细胞仪检测H9C2细胞凋亡水平。结果与模型组比较,agomiR-21组血清炎症因子TNF-α、IL-6、IL-1β水平显著减少(P<0.05),antagomiR-21组则显著增加;与模型组相比,agomiR-21组心肌组织病理变化显著缓解,且心肌细胞凋亡显著减少(P<0.05),miR-21靶基因PDCD4蛋白表达量显著下调(P<0.05),antagomiR-21组心肌组织病变加重,心肌细胞凋亡显著增加(P<0.05),PDCD4蛋白表达量显著上调(P<0.05);体外抑制PDCD4表达后心肌细胞H9C2凋亡显著减少(P<0.05)。结论 miR-21可能通过靶向PDCD4减轻小鼠心脏缺血再灌注损伤。
Objective To study the effect and mechanism of miR-21 on myocardial ischemia-reperfusion injury in mice. Methods C57 BL/6 J mice were modeled as ischemia-reperfusion injury. After modeling,they were divided into control group,model group,agomiR-21 group and antagomiR-21 group. Agmi-21 group and antagomiR-21 group were given miR-21 respectively. The agonist and antagonist,the control group and the model group were given an equal volume of normal saline.After 4 weeks of administration,the expression levels of serum inflammatory factors TNF-α,IL-6 and IL-1β were detected by Elisa;myocardial histopathology was detected by HE staining. The changes of mouse cardiomyocytes were detected by in situ nick end transferase labeling(TUNEL). The relationship between miR-21 and PDCD4 was verified by luciferase reporter assay. The expression of PDCD4 in myocardial tissue of each group was detected by Western blot. The cardiomyocyte H9 C2 was transfected with PDCD4 siRNA in vitro,and then the apoptosis level of H9 C2 cells was detected by flow cytometry.Results The serum inflammatory factors TNF-α,IL-6 and IL-1β in the agomiR-21 group were significantly decreased(P<0.05),and the antagomiR-21 group was significantly increased. The pathological changes of myocardial tissue in the agomiR-21 group were significantly relieved,and the myocardial cells were withered. The expression of PDCR4 was significantly decreased(P<0.05),the expression of PDCD4 was down-regulated(P<0.05),the myocardial tissue of antagomiR-21 group was aggravated,the apoptosis of cardiomyocytes was significantly increased(P<0.05),and the expression of PDCD4 was significant. Up-regulation(P<0.05);H9 C2 apoptosis in cardiomyocytes was significantly decreased after inhibition of PDCD4 expression in vitro(P<0.05). Conclusion The miR-21 may attenuate myocardial ischemia-reperfusion injury in mice by targeting PDCD4.
作者
范丽
姚亚妮
李瑜
FAN Li;YAO Ya-ni;LI Yu(Department of Cadre Ward,First Affiliated Hospital of Xinjiang Medical University,Urumqi,830054,China)
出处
《解剖学研究》
CAS
2019年第5期407-411,共5页
Anatomy Research
基金
新疆维吾尔自治区自然科学基金（2017D01C349）
