miR-323-3p通过靶向TGF-β2/JNK通路影响冠心病大鼠心肌细胞的凋亡-现代医学

miR-323-3p通过靶向TGF-β2/JNK通路影响冠心病大鼠心肌细胞的凋亡

单位：1. 宝鸡市中心医院, 陕西宝鸡 721000;
2. 西安交通大学第二附属医院, 陕西西安 710004

关键词：miR-323-3p 冠心病细胞凋亡转化生长因子β2/c-Jun氨基末端激酶通路大鼠

分类号：R-33;R541.4

出版年·卷·期（页码）：2022·50·第七期（840-846）

摘要：

目的：探讨微小RNA-323-3p（miR-323-3p）对冠心病大鼠心肌细胞凋亡的影响及其可能的作用机制。方法：将SD大鼠分为Control组、Model组、NC agomir组、miR-323-3p agomir组、miR-323-3p agomir+pcDNA组、miR-323-3p agomir+pcDNA-TGF-β2组，每组12只。除Control组外，其它组大鼠均采用高脂饲料喂养以构建冠心病大鼠模型。建模成功后，通过尾静脉注射对应的agomir或pcDNA进行处理，Control组、Model组尾静脉注射等量的生理盐水。2周后，测量大鼠心脏功能指标平均动脉压（MAP）、左室收缩压（LVSP）、心率（HR）的变化；HE染色检测大鼠心肌组织病理变化；TUNEL染色检测各组大鼠心肌细胞凋亡；qRT-PCR检测大鼠心肌组织中miR-323-3p表达；Western blotting检测大鼠心肌组织中Bax、Bcl-2、转化生长因子β2（TGF-β2）及磷酸化c-Jun氨基末端激酶（p-JNK）蛋白表达；双荧光素酶报告基因实验验证miR-323-3p与TGF-β2的关系。结果：与Control组比较，Model组大鼠心肌细胞肿胀，有大量炎症细胞浸润，心肌纤维断裂，MAP、LVSP、HR、miR-323-3p、Bcl-2蛋白表达水平显著降低，细胞凋亡率、Bax、TGF-β2、p-JNK蛋白表达水平显著升高（P<0.05）；与Model组、NC agomir组比较，miR-323-3p agomir组大鼠心肌组织病理改变显著减轻，MAP、LVSP、HR、miR-323-3p、Bcl-2蛋白表达水平显著升高，细胞凋亡率、Bax、TGF-β2、p-JNK蛋白表达水平显著降低（P<0.05）；与miR-323-3p agomir组、miR-323-3p agomir+pcDNA组比较，miR-323-3p agomir+pcDNA-TGF-β2组大鼠心肌组织病理改变显著加重，MAP、LVSP、HR、Bcl-2蛋白表达水平显著降低，细胞凋亡率、Bax、TGF-β2、p-JNK蛋白表达水平显著升高（P<0.05），而miR-323-3p水平变化差异无统计学意义（P>0.05）；miR-323-3p靶向调控TGF-β2的表达。结论：过表达miR-323-3p可能通过抑制TGF-β2/JNK通路抑制冠心病大鼠细胞凋亡。

Objective: To investigate the effect of miRNA-323-3p on cardiomyocyte apoptosis in rats with coronary heart disease and its possible mechanism. Methods: The SD rats were divided into Control group, Model group, NC agomir group, miR-323-3p agomir group, miR-323-3p agomir+pcDNA group, and miR-323-3p agomir+pcDNA-TGF-β2 group, 12 rats in each group. Except for the control group, rats in the other groups were fed with high-fat diet to construct a coronary heart disease rat model. After the modeling was successfully estabilshed, the corresponding agomir or pcDNA was injected into the tail vein for treatment, the control group and the model group were injected with the same amount of normal saline through the tail vein. Two weeks later, the changes in mean arterial pressure(MAP), left ventricular systolic pressure(LVSP), and heart rate(HR) of the rats' heart function indexes were measured; HE staining was used to detect the pathological changes of rat myocardial tissue; TUNEL staining was used to detect myocardial cell apoptosis in rats of each group; qRT-PCR was used to detect the miR-323-3p expression in rat myocardial tissue; Western blotting was used to detect the Bax, Bcl-2, transforming growth factor β2(TGF-β2) and phosphorylated c-Jun N-terminal kinase(p-JNK) protein expression in rat myocardial tissue; and dual luciferase reporter gene experiment was used to verify the relationship between miR-323-3p and TGF-β2. Results: Compared with the control group, the myocardial cells were swollen, with a large number of inflammatory cells infiltrating and myocardial fibers breaking, the MAP, LVSP, HR, and the miR-323-3p and Bcl-2 protein expression levels reduced significantly, and apoptosis rate, Bax, TGF-β2 and p-JNK protein expression levels increased significantly in the model group(P<0.05); compared with the model group and NC agomir group, the pathological changes of myocardial tissue were significantly reduced, the MAP, LVSP, HR, the miR-323-3p and Bcl-2 protein expression levels increased significantly, and apoptosis rate, Bax, the TGF-β2 and p-JNK protein expression levels reduced significantly in the miR-323-3p agomir group(P<0.05); compared with the miR-323-3p agomir group and the miR-323-3p agomir+pcDNA group, the pathological changes of myocardial tissue were significantly worse, the MAP, LVSP, HR, and the Bcl-2 protein expression level reduced significantly, and apoptosis rate, the Bax, TGF-β2 and p-JNK protein expression levels increased significantly in the miR-323-3p agomir+pcDNA-TGF-β2 group(P<0.05), while there was no statistically significant difference in the level of miR-323-3p(P>0.05); in addition, miR-323-3p targeted the regulation of the expression of TGF-β2. Conclusion: Overexpression of miR-323-3p may inhibit apoptosis of coronary heart disease rats by inhibiting the TGF-β2/JNK pathway.

参考文献：

[1] LIN F, CHEN H W, ZHAO G A, et al.Advances in research on the circRNA-miRNA-mRNA network in coronary heart disease treated with traditional Chinese medicine[J].Evid Based Complement Alternat Med, 2020, 2020(1):1-10.
[2] MA L Y, CHEN W W, GAO R L, et al.China cardiovascular diseases report 2018:an updated summary[J].J Geriatr Cardiol, 2020, 17(1):1-8.
[3] XIANG Y, PENG J Q, LIU Q, et al.The association between CD69 and EGR1 levels, and CHD patients without reflow after PCI[J].Exp Ther Med, 2019, 17(5):3913-3920.
[4] YIN H, LIU Y, MA H, et al.Associations of mood symptoms with NYHA functional classes in angina pectoris patients:a cross-sectional study[J].BMC Psychiatry, 2019, 19(1):85-97.
[5] QIN J, SUN Y, LIU S, et al.MicroRNA-323-3p promotes myogenesis by targeting Smad2[J].J Cell Biochem, 2019, 120(11):18751-18761.
[6] ZHAO Y, TAO M, WEI M, et al.Mesenchymal stem cells derived exosomal miR-323-3p promotes proliferation and inhibits apoptosis of cumulus cells in polycystic ovary syndrome(PCOS)[J].Artif Cells Nanomed Biotechnol, 2019, 47(1):3804-3813.
[7] 郭伟伟, 张晓鹏, 李霞, 等.三七皂苷R1调控AMPK/Nrf-2/HO-1信号通路缓解冠心病大鼠心肌损伤的研究[J].中国现代应用药学, 2021, 38(1):36-41.
[8] JIAN W, LI L, WEI X M, et al.Prognostic value of angiopoietin-2 for patients with coronary heart disease after elective PCI[J].Medicine(Baltimore), 2019, 98(5):1-7.
[9] 蒋长荣, 马小峰.SATB1基因沉默介导NF-κB通路对冠心病小鼠主动脉内皮细胞炎症损伤的保护作用[J].东南大学学报(医学版), 2020, 39(1):35-40.
[10] RONG J, XU J, LIU Q, et al.Anti-inflammatory effect of up-regulated microRNA-221-3p on coronary heart disease via suppressing NLRP3/ASC/pro-caspase-1 inflammasome pathway activation[J].Cell Cycle, 2020, 19(12):1478-1491.
[11] TANG L, LIU L, LI G, et al.Expression profiles of long noncoding RNAs in intranasal LPS-mediated alzheimer's disease model in mice[J].Biomed Res Int, 2019, 2019(1):1-14.
[12] YANG X, HE T, HAN S, et al.The role of traditional chinese medicine in the regulation of oxidative stress in treating coronary heart disease[J].Oxid Med Cell Longev, 2019, 2019(1):1-13.
[13] FENG J, WANG M, LI M, et al.Serum miR-221-3p as a new potential biomarker for depressed mood in perioperative patients[J].Brain Res, 2019, 1720(1):146296-146327.
[14] JING R, ZHONG Q Q, LONG T Y, et al.Downregulated miRNA-26a-5p induces the apoptosis of endothelial cells in coronary heart disease by inhibiting PI3K/AKT pathway[J].Eur Rev Med Pharmacol Sci, 2019, 23(11):4940-4947.
[15] SHI C C, PAN L Y, ZHAO Y Q, et al.MicroRNA-323-3p inhibits oxidative stress and apoptosis after myocardial infarction by targeting TGF-β2/JNK pathway[J].Eur Rev Med Pharmacol Sci, 2020, 24(12):6961-6970.
[16] WANG T, LIU Y, LV M, et al.miR-323-3p regulates the steroidogenesis and cell apoptosis in polycystic ovary syndrome(PCOS) by targeting IGF-1[J].Gene, 2019, 683(1):87-100.
[17] MORIKAWA M, DERYNCK R, MIYAZONO K.TGF-β and the TGF-β family:context-dependent roles in cell and tissue physiology[J].Cold Spring Harb Perspect Biol, 2016, 8(5):1-24.
[18] 王凤仙.表没食子儿茶素没食子酸酯通过TGF-β1/JNK信号通路改善糖尿病大鼠心肌纤维化的作用及相关机制[D].锦州:锦州医科大学, 2018.
[19] 杨雅, 刘霞.木补对心肌缺血大鼠ST段及Omentin-1/iNOS和TGF-β1/JNK通路的影响[J].成都医学院学报, 2015, 10(1):31-34.

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