Objective To investigate the role and possible mechanism of miRNA-130a-3p in cardiomyocyte injury induced by LPS. Methods H9C2 cells were divided into the following four groups: a normal control group, a LPS model group, a miRNA negative control group and a miRNA-130a-3p mimics group. The expression level of miRNA-130a-3p mRNA in each group was detected by RT-qPCR. The CCK-8 assay was used to detect the cell activity in each group. The contents of TNF-α, IL-6 and IL-1β were detected by ELISA assay. Western blot was used to detect the protein expression levels of NF-κBp65, IκBα, Bax, Bcl-2 and Cleaved-Caspase-3. Results The results of RT-qPCR showed that the level of miRNA-130a-3p mRNA in the LPS model group was significantly lower than that in normal control group. The results of CCK-8 showed that the cell activity of LPS group was significantly lower than that of normal control group, while compared with the LPS group, the cell activity in miRNA-130a-3p mimics group increased significantly. The results of ELISA showed that the contents of TNF-α, IL-6 and IL-1β in cell culture medium of LPS group were significantly higher than those of normal control group, while compared with the LPS group, the contents of TNF-α, IL-6 and IL-1β in cell culture medium of miRNA-130a-3p mimics group decreased significantly. The results of Western blot showed that compared with normal control group, the protein expression levels of NF-κBp65, Bax and Cleaved-Caspase-3 increased significantly in the LPS group , while the protein expression levels of IκBα and Bcl-2 decreased significantly. Compared with the LPS group, the protein expression levels of NF-κBp65, Bax and Cleaved-Caspase-3 decreased significantly, while the protein expression levels of IκBα and Bcl-2 increased significantly in miRNA-130a-3p mimics group. Conclusions miRNA-130A-3P can inhibit cell apoptosis, improve cell activity and reduce the release of inflammatory factors, so as to alleviate cardiomyocyte injury induced LPS, through inhibiting the activation of NF-κB signaling pathway.
Key words
miRNA-130a-3p; /
/
NF-κB signaling pathway; /
/
Cell injury; /
/
Apoptosis
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
References
[1] Singer M, Deutschman CS, Seymour CW, et al. The third international consensus definitions for sepsis and septic shock (Sepsis-3)[J]. JAMA, 2016, 315(8): 801-810. DOI: 10.1001/jama.2016.0287.
[2] Kolhe R, Hunter M, Liu S, et al. Gender-specific differential expression of exosomal miRNA in synovial fluid of patients with osteoarthritis[J]. Sci Rep, 2017, 7(1): 2029. DOI: 10.1038/s41598-017-01905-y.
[3] 赵轶男, 孙强, 毕龙, 等. miR-130a-3p调控SOX4对骨关节炎软骨细胞增殖、分化和炎症因子释放的影响[J]. 现代生物医学进展, 2019, 19(22): 4201-4207. DOI: 10.13241/j.cnki.pmb.2019.22.001.
[4] 王和杰, 范忠诚, 金旭红, 等. 长链非编码RNA靶向微小RNA130a-3p基因对骨关节炎软骨细胞增殖及炎性因子分泌的影响[J]. 中华实验外科杂志, 2021, 38(5): 871-873. DOI: 10.3760/cma.j.cn421213-20201106-01923.
[5] 许德星, 王伟, 张静怡, 等. miR-130a-3p靶向TRIM37蛋白缓解H2O2诱导的心肌损伤[J]. 华中科技大学学报（医学版）, 2021, 50(1): 14-20. DOI: 10.3870/j.issn.1672-0741.2021.01.003.
[6] 周涛, 杨富瑜, 高士豪, 等. 微小RNA-130a-3p在心肌梗死大鼠心肌组织的表达及其对炎症的影响[J]. 中华实验外科杂志, 2021, 38(8): 1459-1462. DOI: 10.3760/cma.j.cn421213-20210106-01001.
[7] Weiss SL, Asaro LA, Flori HR, et al. Multiple organ dysfunction in children mechanically ventilated for acute respiratory failure[J]. Pediatr Crit Care Med, 2017,18(4): 319⁃329. DOI: 10.1097/PCC.000000 0000001091.
[8] Zhang N, Feng H, Liao HH, et al. Myricetin attenuated LPS induced cardiac injury in vivo and in vitro[J]. Phytother Res, 2018, 32(3): 459-470. DOI: 10.1002/ptr.5989.
[9] 刘安雷, 刘聚源, 徐军, 等. 交感神经兴奋与树突状细胞活化加剧脂多糖诱导大鼠心肌损伤[J]. 基础医学与临床, 2018, 38(7): 973-977. DOI: 10.3969/j.issn.1001-6325.2018.07.014.
[10] 雷家秀, 朱雅萍, 张雨. 依托咪酯上调miR-290-5p减轻LPS诱导的大鼠心肌细胞系H9C2 损伤[J]. 基础医学与临床, 2021, 41(6): 848-853. DOI: 10.3969/j.issn.1001-6325.2021.06.012.
[11] 邢若, 王鹏, 李兴杰. 山药多糖对LPS诱导的心肌H9C2细胞炎症因子表达和细胞凋亡的影响及机制[J]. 中国免疫学杂志, 2021, 37(6): 683-688. DOI: 10.3969/j.issn.1000-484X.2021.06.009.
[12]雷肖蠢, 张海良, 丁鹏. LncRNA MFI2-AS1靶向miR-331-3p调节LPS诱导的心肌细胞损伤[J]. 河北医药, 2021, 43(8): 1142-1146. DOI: 10.3969/j.issn.1002-7386.2021.08.004.
[13]Huang ZQ, Xu W, Wu JL, et al. MicroRNA-374a protects against myocardial ischemia-reperfusion injury in mice by targeting the MAPK6 pathway[J]. Life Sci, 2019, 232: 116619. DOI: 10.1016/j.lfs.2019.116619.
[14] Shi J, Bei Y, Kong X, et al. miR-17-3p contributes to exercise-induced cardiac growth and protects against myocardial ischemia-reperfusion injury[J]. Theranostics, 2017, 7(3): 664-676. DOI: 10.7150/thno.15162.
[15] Ren Q, Zhao S, Ren C, et al. Astragalus polysaccharide alleviates LPS-induced inflammation injury by regulating miR-127 in H9c2 cardiomyoblasts[J]. Int J Immunopathol Pharmacol, 2018, 32: 2058738418759180. DOI: 10.1177/2058738418759180.
[16] Wang L, Zhang Y, Zhu G, et al. miR-16 exhibits protective function in LPS-treated cardiomyocytes by targeting DOCK2 to repress cell apoptosis and exert anti-inflammatory effect[J]. Cell Biol Int, 2020, 44(8): 1760-1768. DOI: 10.1002/cbin.11371.
[17]王晓姣, 曲径, 李东旭, 等. 过表达miR-130a-3p对心肌细胞肥大的缓解作用研究[J].生物医学工程杂志, 2020, 37(2): 340-348. DOI: 10.7507/1001-5515.201911049
[18] Lawrence T. The nuclear factor NF-kappaB pathway in inflammation[J]. Cold Spring Harb Perspect Biol, 2009, 1(6): a001651. DOI: 10.1101/cshperspect.a001651.
[19] Balzano T, Arenas YM, Dadsetan S, et al. Sustained hyperammonemia induces TNF-a IN Purkinje neurons by activating the TNFR1-NF-κB pathway[J]. J Neuroinflammation, 2020, 17(1): 70. DOI: 10.1186/s12974-020-01746-z.
[20]Zusso M, Lunardi V, Franceschini D, et al. Ciprofloxacin and levofloxacin attenuate microglia inflammatory response via TLR4/NF-kB pathway[J]. J Neuroinflammation, 2019, 16(1): 148. DOI: 10.1186/s12974-019-1538-9.
[21] Wei JL, Wu CJ, Chen JJ, et al. LncRNA NEAT1 promotes the progression of sepsis-induced myocardial cell injury by sponging miR-144-3p[J]. Eur Rev Med Pharmacol Sci, 2020, 24(2): 851⁃861. DOI: 10.26355/eurrev_202001_20069.
[22] Papathanasiou I, Balis C, Trachana V, et al. The synergistic function of miR-140-5p and miR-146a on TLR4-mediated cytokine secretion in osteoarthritic chondrocytes[J]. Biochem Biophys Res Commun, 2020, 522(3): 783⁃791. DOI: 10.1016/j.bbrc.2019.11.168.
[23] Yang Y, Liu DD, Xi Y, et al. Upregulation of miRNA-140-5p inhibits inflammatory cytokines in acute lung injury through the MyD88/NF⁃κB signaling pathway by targeting TLR4[J]. Exp Ther Med, 2018, 16(5): 3913⁃3920. DOI: 10.3892/etm.2018.6692.
[24] 王成, 金卫东, 郭长磊, 等. 沉默MyD88抑制NF-κB p65活化对TNF-α诱导的心肌细胞凋亡和炎症损伤的保护作用[J]. 现代免疫学, 2021, 41(3): 204-209. DOI: 1001-2478(2021)03-0204-06.
[25] 王可, 董平栓, 和素娜, 等. miR-199a在LPS诱导的大鼠原代心肌细胞中通过激活NF-κB信号通路加重心肌细胞损伤[J]. 中国比较医学杂志,2020, 30(10):63-69. DOI: 10.3969/j.issn.1671-7856. 2020. 10. 009.
PDF(3915 KB)
