环状RNA_0124644调控微小RNA-136对高糖诱导的肾小管上皮细胞损伤的影响  

Circular RNA_0124644 regulates microRNA-136 on renal tubular epithelial cells damage induced by high glucose

作　　者：蔡莉 侯保健 贺映侠 CAI Li;HOU Baojian;HE Yingxia(Department of Endocrinology,Third People s Hospital of Hubei Province,Wuhan 430000,China;Department of General Medicine,Wuhan Central Hospital,Wuhan 430000,China)

机构地区：[1]湖北省第三人民医院内分泌科,武汉430000 [2]武汉市中心医院全科医学科

出　　处：《肾脏病与透析肾移植杂志》2024年第5期444-450,共7页Chinese Journal of Nephrology,Dialysis & Transplantation

摘　　要：目的:探讨环状RNA_0124644(circ_0124644)对高糖(HG)诱导的人肾小管上皮细胞(HK-2细胞)损伤的影响。方法:收集29例糖尿病肾病患者及41例健康体检者的血浆,实时荧光定量PCR(qRT-PCR)检测circ_0124644和微小RNA(miR)-136的表达;验证circ_0124644和miR-136的靶向关系。将HK-2细胞随机分为正常糖(NG)组、HG组、HG+si-circ_0124644组、HG+si-NC组、HG+si-circ_0124644+anti-miR-136组、HG+si-circ_0124644+anti-miR-NC组。甲基噻唑基四唑(MTT)法检测细胞抑制率;检测细胞凋亡率和活化的半胱氨酸蛋白酶3(Cleaved caspase-3)蛋白水平以及白细胞介素(IL)-6、IL-10、IL-1β、超氧化物歧化酶(SOD)、乳酸脱氢酶(LDH)、丙二醛(MDA)、活性氧(ROS)水平。结果:与健康体检者相比,糖尿病肾病患者血浆中circ_0124644表达升高,miR-136表达降低(P<0.05)。HG可诱导的HK-2细胞中circ_0124644表达升高,miR-136表达降低,细胞抑制率、凋亡率、Cleaved caspase-3蛋白、Cleaved caspase-3平均荧光强度、IL-6、IL-1β、LDH和MDA水平及ROS活性升高,IL-10水平和SOD活性降低(P<0.05)。沉默circ_0124644后,HK-2细胞抑制率、凋亡率和Cleaved caspase-3蛋白表达、Cleaved caspase-3平均荧光强度降低,IL-6、IL-1β水平降低,IL-10水平升高,SOD活性升高,LDH水平和MDA含量及ROS活性降低(P<0.05)。下调miR-136逆减弱了沉默circ_0124644对HG诱导的HK-2细胞的作用。结论:沉默circ_0124644可能通过调控miR-136抑制HG诱导的肾小管上皮细胞损伤。Objective:To investigate the effect of circular RNA_0124644(circ_0124644)on human renal tubular epithelial cells(HK-2 cells)damage induced by high glucose(HG).Methodology:The plasma of 29 diabetic nephropathy patients and 41 healthy subjects were collected.The expression of circ_0124644 and microrna(Mir-136)were detected by real-time quantitative fluorescent PCR(qRT-PCR);the targeting relationship between circ_0124644 and microRNA(miR)-136 was verified.HK-2 cells were randomly divided into normal glucose(NG)group,HG group,HG+si-circ_0124644 group,HG+si-NC group,HG+si-circ_0124644+anti-miR-136 group,HG+si-circ_0124644+anti-miR-NC group.The cell inhibitory rate was detected by MTT assay;apoptosis rate,Cleaved caspase-3 protein levels,interleukin(IL-6),IL-10,IL-1β,superoxide dismutase(SOD),lactate dehydrogenase(LDH),malondialdehyde(MDA),and reactive oxygen species(ROS)levels were measured.Results:Compared with the healthy subjects,the expression of circ_0124644 in the plasma of diabetic nephropathy patients was increased,and the expression of miR-136 was decreased(P<0.05).In HG-induced HK-2 cells,the expression level of circ_0124644 was increased,the expression of miR-136 was decreased,the cell inhibitory rate,apoptosis rate,the levels of Cleaved caspase-3 protein,average fluorescence intensity of Cleaved caspase-3,IL-6,IL-1β,LDH,MDA,and activity of ROS were increased,level of IL-10 and activity of SOD were decreased(P<0.05).After silencing circ_0124644,HK-2 cell inhibition rate,apoptosis rate,expression level of Cleaved caspase-3 protein and average fluorescence intensity of Cleaved caspase-3 were decreased,levels of IL-6,IL-1βwere decreased,level of IL-10 was increased,activity of SOD was increased,level of LDH,content of MDA and activity of ROS were decreased(P<0.05).Down-regulation of miR-136 reversely weakened the effect of silencing circ_0124644 on HK-2 cells induced by HG.Conclusion:Silencing circ_0124644 may inhibit HG-induced renal tubular epithelial cells injury by regulating miR-136.

关 键 词：糖尿病肾病 环状RNA_0124644 微小RNA-136 肾小管上皮细胞 炎症因子 氧化应激 凋亡 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]