Zhu Bei, Cheng Xingbo, Jiang Yilan, Cheng Ming, Chen Luping, Bao Jiajun, Tang Xiaofeng
Department of Endocrinology, The First Affiliated Hospital of Soochow University, Suzhou 215006, People's Republic of China, Department of Endocrinology, Rugao People's Hospital, Nantong 226500, People's Republic of China.
Department of Endocrinology, The First Affiliated Hospital of Soochow University, Suzhou 215006, People's Republic of China.
Diabetes Metab Syndr Obes. 2020 Feb 13;13:365-375. doi: 10.2147/DMSO.S225791. eCollection 2020.
Long noncoding RNAs (lncRNAs) can regulate the progression of DN. This research aimed to study the effect of lncRNA KCNQ1OT1 on the oxidative stress and pyroptosis of the renal tubular epithelial cells induced by high glucose (HG).
RT-qPCR analysis detected the KCNQ1OT1 expression in serum with DN and HG-induced HK-2 cells, detect the expression of NLRP3, cleaved-caspase1, P-caspase1, IL-1β, p-IL-1β and GSDMD-N in HG-induced HK-2 cells, and confirm the transfection effects. The expression of NLRP3, cleaved-caspase1, P-caspase1, IL-1β, p-IL-1β and GSDMD-N in HG-induced HK-2 cells was also analyzed by Western blot analysis. ELISA assay detected the levels of TNF-α, IL-6 and MCP-1. The levels of ROS, MDA and SOD were determined by respective ELISA kits and ROS was also detected by the ROS assay kit (containing DCFH-DA).
We found that KCNQ1OT1 was increased in the plasma of patients with DN and HG-induced HK-2 cells and KCNQ1OT1 interference could decrease the inflammation, oxidative stress and pyroptosis of HG-induced HK-2 cells. In addition, KCNQ1OT1 directly targets miR-506-3p. MiR-506-3p was downregulated in the plasma of patients with DN and HG-induced HK-2 cells and KCNQ1OT1 interference promoted the expression of miR-506-3p. MiR-506-3p overexpression suppressed the inflammation, oxidative stress and pyroptosis of HG-induced HK-2 cells.
This study demonstrated that downregulation of KCNQ1OT1 inhibited the inflammation, oxidative stress and pyroptosis of HG-induced HK-2 cells by up-regulating the expression of miR-506-3p, which provide new insights into the treatment of DN.
长链非编码RNA（lncRNA）可调节糖尿病肾病（DN）的进展。本研究旨在探讨lncRNA KCNQ1OT1对高糖（HG）诱导的肾小管上皮细胞氧化应激和焦亡的影响。
采用RT-qPCR分析检测DN患者血清及HG诱导的HK-2细胞中KCNQ1OT1的表达，检测HG诱导的HK-2细胞中NLRP3、裂解型半胱天冬酶-1、磷酸化半胱天冬酶-1、白细胞介素-1β、磷酸化白细胞介素-1β和Gasdermin D-N的表达，并确认转染效果。采用蛋白质免疫印迹分析检测HG诱导的HK-2细胞中NLRP3、裂解型半胱天冬酶-1、磷酸化半胱天冬酶-1、白细胞介素-1β、磷酸化白细胞介素-1β和Gasdermin D-N的表达。酶联免疫吸附测定（ELISA）检测肿瘤坏死因子-α、白细胞介素-6和单核细胞趋化蛋白-1的水平。采用相应的ELISA试剂盒测定活性氧（ROS）、丙二醛（MDA）和超氧化物歧化酶（SOD）的水平，并用ROS检测试剂盒（含2',7'-二氯二氢荧光素二乙酸酯）检测ROS。
我们发现，DN患者血浆及HG诱导的HK-2细胞中KCNQ1OT1升高，KCNQ1OT1干扰可减轻HG诱导的HK-2细胞的炎症、氧化应激和焦亡。此外，KCNQ1OT1直接靶向miR-506-3p。miR-506-3p在DN患者血浆及HG诱导的HK-2细胞中表达下调，KCNQ1OT1干扰可促进miR-506-3p的表达。miR-506-3p过表达可抑制HG诱导的HK-2细胞的炎症、氧化应激和焦亡。
本研究表明，下调KCNQ1OT1可通过上调miR-506-3p的表达抑制HG诱导的HK-2细胞的炎症、氧化应激和焦亡，为DN的治疗提供了新的思路。
