CIRC_0044235靶向miR-574-5p减轻IL-1β诱导的人软骨细胞损伤  

CIRC_0044235 targeting miR-574-5p attenuates IL-1β-induced injury in human chondrocyte

作　　者：汪博 张明焕 WANG Bo;ZHANG Minghuan(Department of Anesthesiology,Wuhan Third Hospital,Tongren Hospital Affiliated to Wuhan University,Wuhan 430071,China;Department of Orthopedics,Wuhan Third Hospital,Tongren Hospital Affiliated to Wuhan University,Wuhan 430071,China)

机构地区：[1]武汉市第三医院武汉大学附属同仁医院麻醉科,湖北武汉430071 [2]武汉市第三医院武汉大学附属同仁医院骨科,湖北武汉430071

出　　处：《基础医学与临床》2023年第3期419-426,共8页Basic and Clinical Medicine

基　　金：2021年武汉市医学科研项目(WX21A01)。

摘　　要：目的观察CIRC_0044235是否靶向miR-574-5p影响白细胞介素(IL)-1β诱导的软骨细胞凋亡、氧化应激及炎性因子表达。方法将细胞分为对照组、IL-1β组、分别转染pcDNA、pcDNA-CIRC_0044235、anti-miR-NC、anti-miR-574-5p、pcDNA-CIRC_0044235+miR-NC和pcDNA-CIRC_0044235+miR-574-5p,干预IL-1β。RT-qPCR检测细胞中CIRC_0044235和miR-574-5p表达。流式细胞测量术与Western blot分别检测细胞凋亡、相关蛋白Bcl-2与Bax的表达;比色法检测超氧化物歧化酶(SOD)和乳酸脱氢酶(LDH)活力;ELISA检测IL-6和IL-10的水平。双荧光素酶报告实验检测CIRC_0044235和miR-574-5p的靶向结合。结果与对照组相比,IL-1β组软骨细胞中CIRC_0044235表达量、Bcl-2蛋白表达量、SOD活力和IL-10含量降低(P<0.05),而miR-574-5p表达量、凋亡率、Bax蛋白表达量、LDH活力和IL-6含量升高(P<0.05);与IL-1β组或pcDNA干预组相比,pcDNA-CIRC_0044235组软骨细胞中CIRC_0044235表达量、Bcl-2蛋白表达量、SOD活力和IL-10含量增多,而miR-574-5p表达量、凋亡率、Bax蛋白表达量、LDH活力和IL-6含量减少(P<0.05)。CIRC_0044235靶向调控miR-574-5p。anti-miR-574-5p组软骨细胞中miR-574-5p表达量、凋亡率、Bax蛋白表达量、LDH活力和IL-6含量比anti-miR-NC组减少,Bcl-2蛋白表达量、SOD活力与IL-10含量比anti-miR-NC组增加(P<0.05)。转染pcDNA-CIRC_0044235+miR-574-5p组软骨细胞中miR-574-5p表达量、凋亡率、Bax蛋白表达量、LDH活力和IL-6含量高于pcDNA-CIRC_0044235+miR-NC组,Bcl-2蛋白表达量、SOD活力及IL-10含量低于pcDNA-CIRC_0044235+miR-NC组(P<0.05)。结论CIRC_0044235通过靶向miR-574-5p,抑制IL-1β诱导的人关节软骨细胞凋亡、氧化应激和炎性因子表达。Objective To investigate whether CIRC_0044235 targeting miR-574-5p could affect the levels of chondro-cyte apoptosis,oxidative stress and inflammatory factor expression induced by interleukin(IL)-1β.Methods The cells were divided into control group,IL-1βgroup,transfected with pcDNA,pcDNA-CIRC_0044235,anti-miR-NC,anti-miR-574-5p,pcDNA-CIRC_0044235+miR-NC,pcDNA-CIRC_0044235+miR-574-5p,interfere with IL-1β.RT-qPCR was used to detect the expression of CIRC_0044235 and miR-574-5p in cell.Flow cytometry and Western blot were used to detect apoptosis and the expression of related proteins Bcl-2 and Bax,respectively.Colorimetry was used to superoxide dismutase(SOD)and lactate dehydrogenase(LDH)activity were detected,and ELISA was used to detect the levels of IL-6 and IL-10.A dual-luciferase reporter assay was used for examining the targeted binding of CIRC_0044235and miR-574-5p.Results Compared with the control group,the expression of CIRC_0044235,the expression of Bcl-2 protein,the activity of SOD and the content of IL-10 in chondrocytes in the IL-1βgroup were decreased(P<0.05),while the expression of miR-574-5p and the rate of apoptosis were decreased(P<0.05).Bax protein expression,LDH activity and IL-6 content all increased(P<0.05).Compared with IL-1βgroup or pcDNA intervention group,CIRC_0044235 expression,Bcl-2 protein expression,SOD activity and IL-10 content in chondrocytes of pcDNA-CIRC_0044235 group increased,while miR-574-5p expression,apoptosis rate,Bax protein expression,LDH activity and IL-6 content decreased(P<0.05).CIRC_0044235 targeted miR-574-5p.miR-574-5p expression,apoptosis rate,Bax protein expression,LDH activity and IL-6 content in chondrocytes of anti-miR-574-5p group were all lower than those found in anti-miR-NC group.Bcl-2 protein expression,SOD activity and IL-10 content were higher than those in anti-miR-NC group(P<0.05).miR-574-5p expression,apoptosis rate,Bax protein expression,LDH activity and IL-6 content in chondrocytes of pcDNA-CIRC_0044235+miR-574-5p group were higher than those of

关 键 词：骨关节炎 CIRC_0044235 miR-574-5p 凋亡 氧化应激 

分 类 号：R684.3[医药卫生—骨科学]