天麻素通过调控LINC01619/miR-216a-5p对高糖诱导的肾小管上皮细胞HK-2损伤的影响

Effect of Gastrodin on High Glucose-induced HK-2 Injury by Regulating LINC01619/miR-216a-5p

目的探讨天麻素对高糖诱导的肾小管上皮细胞(HK-2)损伤的影响及其机制。方法体外培养HK-2细胞分为正常糖(NG)组、高糖(HG)组、HG+天麻素低剂量组、HG+天麻素中剂量组、HG+天麻素高剂量组、HG+pcDNA组、HG+pcDNA-LINC01619组、HG+天麻素高剂量+si-NC组、HG+天麻素高剂量+si-LINC01619组。实时定量PCR(qRT-PCR)检测LINC01619和miR-216a-5p表达。CCK-8法和流式细胞术检测HK-2细胞活力和凋亡率。试剂盒检测丙二醛(MDA)含量以及超氧化物歧化酶(SOD)和乳酸脱氢酶(LDH)活性。双荧光素酶报告基因实验验证LINC01619和miR-216a-5p靶向关系。结果高糖诱导后HK-2细胞凋亡率、miR-216a-5p表达量、MDA含量、LDH活性显著升高(均P<0.05),SOD活性、LINC01619表达量显著降低(均P<0.05)。中、高剂量的天麻素显著减弱高糖诱导后HK-2细胞凋亡(P<0.05),降低MDA含量、LDH活性、miR-216a-5p表达量(均P<0.05),并增加SOD活性、LINC01619表达量(均P<0.05)。过表达LINC01619显著减弱高糖诱导后HK-2细胞凋亡(P<0.05),降低MDA含量、LDH活性、miR-216a-5p表达量(均P<0.05),并增加SOD活性(P<0.05)。干扰LINC01619表达能够减弱高剂量天麻素对高糖诱导的HK-2细胞凋亡和氧化损伤的影响(均P<0.05)。结论天麻素能够减弱高糖诱导的肾小管上皮细胞HK-2凋亡和氧化损伤,其机制可能与调控LINC01619/miR-216a-5p通路有关。

Objective To investigate the effect of gastrodin on the injury of renal tubular epithelial cells(HK-2)induced by high glucose and its mechanism.Methods HK-2 cells cultured in vitro were divided into normal glucose(NG)group,high glucose(HG)group,HG+low dose gastrodin group,HG+medium dose gastrodin group,HG+high dose gastrodin group,HG+pcDNA group,HG+pcDNA-LINC01619 group,HG+high dose gastrodin+si-NC group,and HG+high dose gastrodin+si-LINC01619 group.Real-time quantitative PCR(qRT-PCR)was used to detect the expression of LINC01619 and miR-216 a-5 p.CCK-8 method and flow cytometry were conducted to detect HK-2 cell viability and apoptosis rate.The testing kit was used to detect the content of malondialdehyde(MDA),the activity of superoxide dismutase(SOD)and lactate dehydrogenase(LDH).The dual luciferase reporter experiment was applied to verify the targeting relationship between LINC01619 and miR-216 a-5 p.Results HK-2 cell apoptosis rate,miR-216 a-5 p expression,MDA content,and LDH activity were significantly increased(all P<0.05).SOD activity and LINC01619 expression were significantly decreased after treatment with high glucose(both P<0.05).Medium and high dose of gastrodin significantly reduced high glucose-induced HK-2 cells apoptosis,MDA content,LDH activity and miR-216 a-5 p expression(all P<0.05),while increased SOD activity and LINC01619 expression(both P<0.05).LINC01619 overexpression significantly reduced high glucose-induced HK-2 cell apoptosis,MDA content,LDH activity and miR-216 a-5 p expression(all P<0.05),while increased SOD activity(P<0.05).Interference with LINC01619 expression could attenuate the effect of high dose gastrodin on the high glucose-induced apoptosis and oxidative damage of HK-2 cell(P<0.05).Conclusion Gastrodin could attenuate high glucose-induced apoptosis and oxidative damage of renal tubular epithelial cell HK-2.The mechanism might be related to the regulation of the LINC01619/miR-216 a-5 p pathway.