circ_0003204靶向miR-346对高糖诱导肾小管上皮细胞损伤的影响-现代医学

circ_0003204靶向miR-346对高糖诱导肾小管上皮细胞损伤的影响

单位：朝阳市第二医院内分泌科, 辽宁朝阳 122000

关键词：糖尿病肾病 circ_0003204 miR-346 细胞凋亡氧化应激炎症

分类号：R587.2;R363.2

出版年·卷·期（页码）：2022·50·第七期（847-853）

摘要：

目的：探讨circ_0003204对高糖（HG）诱导肾小管上皮细胞损伤的影响及其机制。方法：体外培养肾小管上皮细胞（HK-2细胞），经30 mmol·L^-1 HG处理24 h后，qRT-PCR法检测细胞中circ_0003204和miR-346表达量。双荧光素酶报告基因实验验证circ_0003204和miR-346的调控关系。分别转染circ_0003204小干扰RNA、miR-346模拟物或共转染circ_0003204小干扰RNA和miR-346抑制剂至HK-2细胞中，然后用30 mmol·L^-1 HG处理24 h，CCK-8法检测细胞增殖抑制率，流式细胞仪检测细胞凋亡，试剂盒检测细胞培养上清液中乳酸脱氢酶（LDH）、白细胞介素（IL）-6和IL-10水平及细胞中活性氧（ROS）活性、超氧化物歧化酶（SOD）活性、丙二醛（MDA）含量。结果：HK-2细胞经HG处理后，细胞中circ_0003204表达量升高（P<0.05），miR-346表达量降低（P<0.05）。circ_0003204在HK-2细胞中靶向负调控miR-346。干扰circ_0003204或过表达miR-346的HK-2细胞经HG处理后，细胞增殖抑制率和凋亡率降低（P<0.05），培养上清液中LDH、IL-6水平及细胞中ROS活性、MDA含量降低（P<0.05），培养上清液中IL-10水平和细胞中SOD活性升高（P<0.05）。干扰miR-346逆转了干扰circ_0003204对HG诱导的HK-2细胞凋亡、氧化应激和炎症因子表达的影响。结论：干扰circ_0003204可能通过靶向上调miR-346抑制HG诱导的HK-2细胞凋亡、氧化应激和炎症反应。

Objective: To investigate the effect of circ_0003204 on renal tubular epithelial cell injury induced by high glucose(HG) and its mechanism. Methods: The renal tubular epithelial cells HK-2 were cultured in vitro and treated with 30 mmol·L^-1 HG for 24 h, and then the expressions of circ_0003204 and miR-346 in the cells were detected by qRT-PCR. The dual luciferase reporter gene experiment verified the regulatory relationship between circ_0003204 and miR-346. After HK-2 cells transfected with circ_0003204 small interfering RNA or miR-346 mimic, or co-transfected with circ_0003204 small interfering RNA and miR-346 inhibitor, respectively, they were treated with 30 mmol·L^-1 HG for 24 h. And then CCK-8 method was used to detect the inhibition rate of cell proliferation. Flow cytometry was used to detect cell apoptosis. The kits were used to detect the content of lactate dehydrogenase(LDH), interleukin(IL)-6 and IL-10 in cell culture supernatant and the activity of reactive oxygen(ROS), superoxide dismutase(SOD) and the content of malonaldehyde(MDA) in the cells. Results: After HK-2 cells treated with HG, the expression of circ_0003204 in the cells was increased(P<0.05), but the expression of miR-346 was decreased(P<0.05). circ_0003204 negatively regulated the expression of miR-346 in HK-2 cells. After the HK-2 cells that interfered with circ_0003204 or overexpressed miR-346 were treated with HG, the cell proliferation inhibition rate and apoptosis rate were reduced(P<0.05), the content of LDH and IL-6 in the culture supernatant, and the activity of ROS and the content of MDA in the cells were reduced(P<0.05), but the content of IL-10 in the culture supernatant and the activity of SOD in cells were increased(P<0.05). Interfering with miR-346 reversed the effect of interfering with circ_0003204 on HG-induced HK-2 cell apoptosis, oxidative stress and inflammatory factor expression. Conclusion: Interfering with circ_0003204 may inhibit HG-induced HK-2 cell apoptosis, oxidative stress and inflammation by targeting up-regulation of miR-346.

参考文献：

[1] 韩昕健, 郝伟, 孙秀燕, 等.CD36在糖尿病肾病患者肾组织中的表达及调控Wnt/β-catenin信号通路对人肾小管上皮细胞增殖凋亡的实验研究[J].东南大学学报(医学版), 2017, 36(5):704-710.
[2] WAN H, YOU T, LUO W.circ_0003204 regulates cell growth, oxidative stress, and inflammation in ox-LDL-induced vascular endothelial cells via regulating miR-942-5p/HDAC9 axis[J/OL].Front Cardiovasc Med, 2021, 8.doi:10.3389/fcvm.2021.646832.
[3] HUANG X B, SONG K J, CHEN G B, et al.Circular RNA hsa_circ_0003204 promotes cervical cancer cell proliferation, migration, and invasion by regulating MAPK pathway[J].Cancer Biol Ther, 2020, 21(10):972-982.
[4] SU Q X, DONG X H, TANG C H, et al.Knockdown of circ_0003204 alleviates oxidative low-density lipoprotein-induced human umbilical vein endothelial cells injury:circulating RNAs could explain atherosclerosis disease progression[J].Open Med(Wars), 2021, 16(1):558-569.
[5] ZHANG Y, XIAO H Q, WANG Y, et al.Differential expression and therapeutic efficacy of microRNA-346 in diabetic nephropathy mice[J].Exp Ther Med, 2015, 10(1):106-112.
[6] 王殿云, 卢聪聪, 范国霞.桃叶珊瑚苷通过调控miR-30a-5p表达对高糖诱导的肾小管上皮细胞损伤的影响[J].中国免疫学杂志, 2021, 37(4):415-420.
[7] 吴青华, 李冰涛, 涂珺.复方中药治疗糖尿病的研究进展[J].中国中药杂志, 2019, 44(6):1104-1109.
[8] 杨洋, 董敏, 韩滨, 等.溪黄草水提物对高糖诱导的大鼠HBZY-1细胞炎症反应和氧化应激的抑制作用以及对TLR4/NF-κB/NLRP3通路的作用[J].中国医院药学杂志, 2019, 39(21):2157-2161.
[9] 华欣, 孙岳, 朱晓莉.环状RNA在肺腺癌中的差异表达分析[J].东南大学学报(医学版), 2019, 38(6):1019-1023.
[10] YANG H, WANG H, SHANG H, et al.Circular RNA circ_0000950 promotes neuron apoptosis, suppresses neurite outgrowth and elevates inflammatory cytokines levels via directly sponging miR-103 in Alzheimer's disease[J].Cell Cycle, 2019, 18(18):2197-2214.
[11] SUN B, LIU X X, PENG H, et al.Circular RNA _NLRP1 targets mouse microRNA-199b-3p to regulate apoptosis and pyroptosis of hippocampal neuron under oxygen-glucose deprivation exposure[J].Bioengineered, 2021, 12(1):3455-3466.
[12] XIAO L, LUO D, PI P, et al.Up-regulation of miR-135b expression induced by oxidative stress promotes the apoptosis of renal tubular epithelial cells under high glucose condition[J].Clin Exp Pharmacol Physiol, 2020, 47(8):1410-1419.
[13] ZHAO C S, YANG F, WEI X N, et al.miR-139-5p upregulation alleviated spontaneous recurrent epileptiform discharge-induced oxidative stress and apoptosis in rat hippocampal neurons via regulating the Notch pathway[J].Cell Biol Int, 2021, 45(2):463-476.
[14] CHEN H M, JIN G J.Downregulation of Salusin-β protects renal tubular epithelial cells against high glucose-induced inflammation, oxidative stress, apoptosis and lipid accumulation via suppressing miR-155-5p[J].Bioengineered, 2021, 12(1):6155-6165.
[15] 孟斌, 阎成全, 才胜勇.雷公藤多苷对糖尿病肾病大鼠肾组织的保护作用研究[J].中国临床药理学杂志, 2020, 36(14):1270-1273.
[16] XIAO J, LIU D, JIAO W J, et al.Effects of microRNA-346 on epithelial-mesenchymal transition in mouse podocytes[J].Gene, 2015, 560(2):195-199.
[17] ZHENG X W, LI J X, FAN Q, et al.Dexmedetomidine alleviates myocardial ischemia/reperfusion-induced injury and Ca overload via the microRNA-346-3p/CaMKIId axis[J].Int J Cardiol, 2021, 338:185-195.

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