miR-214-3p调控PI3K/AKT通路对软骨细胞自噬和凋亡的影响  

Effects of miR-214-3p on autophagy and apoptosis of chondrocytes through regulation of PI3K/AKT pathway

作　　者：付亚静 雷豫 张莉莹 刘超 汪伟 FU Yajing;LEI Yu;ZHANG Liying;LIU Chao;WANG Wei(Department of Anesthesiology,People’s Hospital of Wuhan University,Wuhan 430000,China;Department of Spinal Orthopedics,the Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine,Guiyang 550003,China;School of Acupuncture and Bone Injury,Hubei University of Chinese Medicine,Wuhan 430065,China;Department of Orthopedics and Traumatology,the Affiliated Hospital of Hubei University of Chinese Medicine,Hubei Provincial Hospital of Traditional Chinese Medicine,Hubei Provincial Institute of Traditional Chinese Medicine,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院麻醉科,湖北武汉430000 [2]贵州中医药大学第二附属医院脊柱骨科,贵州贵阳550003 [3]湖北中医药大学针灸骨伤学院,湖北武汉430065 [4]湖北中医药大学附属医院/湖北省中医院/湖北省中医药研究院骨伤科,湖北武汉430060

出　　处：《中国骨质疏松杂志》2024年第11期1561-1567,共7页Chinese Journal of Osteoporosis

基　　金：湖北省自然科学基金(2021CFB217)。

摘　　要：目的探讨miR-214-3p对IL-1β诱导的软骨细胞自噬和凋亡的影响及其可能的机制。方法采用10 ng/mL的IL-1β诱导大鼠关节软骨细胞24 h,构建骨关节炎(osteoarthritis,OA)细胞模型,将细胞分为对照组、模型组、miR-NC组、miR-214-3p mimics组、740Y-P(PI3K/AKT通路激活剂)+miR-214-3p mimics组和LY294002(PI3K/AKT通路抑制剂)+miR-214-3p mimics组。通过CCK8检测细胞活力、流式细胞术检测细胞凋亡、透射电镜观察细胞中的自噬小体、Western blot检测凋亡(Bcl-2、Bax)、自噬(LC3II、Beclin-1)和PI3K/AKT通路(AKT、p-AKT)相关蛋白的表达。结果与对照组相比,模型组细胞活力降低(P<0.01),细胞凋亡率升高(P<0.01),自噬小体明显减少,细胞中Bcl-2、LC3II和Beclin-1蛋白表达水平降低(P<0.01),Bax和p-AKT/AKT水平升高(P<0.01);与miR-NC组相比,miR-214-3p mimics组细胞活力升高(P<0.01),细胞凋亡率降低(P<0.01),自噬小体明显增多,细胞中Bcl-2、LC3II和Beclin-1蛋白表达水平升高(P<0.01),Bax和p-AKT/AKT水平降低(P<0.01);与miR-214-3p mimics单独作用相比,联合740Y-P能够逆转miR-214-3p mimics的上述作用,而联合LY294002能够进一步加重miR-214-3p mimics的上述作用。结论miR-214-3p能够促进OA细胞增殖和自噬,并抑制细胞凋亡,其机制可能与抑制PI3K/AKT通路的激活有关。Objective To investigate the effects of miR-214-3p on chondrocyte autophagy and apoptosis induced by IL-1βand its possible mechanism.Methods Rat articular chondrocytes were induced by 10 ng/mL IL-1βfor 24 h,to establish osteoarthritis(OA)cell model.The cells were divided into control group,model group,miR-NC group,miR-214-3p mimics group,740Y-P(PI3K/AKT pathway activator)+miR-214-3p mimics group,and LY294002(PI3K/AKT pathway inhibitor)+miR-214-3p mimics group.Cell viability was detected with CCK8.Cell apoptosis was detected with flow cytometry.Autophagosomes in cells were observed using transmission electron microscopy.Expressions of apoptosis(Bcl-2,Bax),autophagy(LC3II,Beclin-1),and PI3K/AKT pathway(AKT,p-AKT)related proteins were detected using Western blotting.Results Compared to those in the control group,the cell viability in the model group decreased(P<0.01),the apoptosis rate increased(P<0.01),autophagosomes decreased,the expression levels of Bcl-2,LC3II,and Beclin-1 proteins decreased(P<0.01),and Bax and p-AKT/AKT levels increased(P<0.01).Compared to those in the miR-NC group,cell viability in miR-214-3p mimics group increased(P<0.01),the apoptosis rate decreased(P<0.01),autophagosomes increased,the expression levels of Bcl-2,LC3II and Beclin-1 proteins increased(P<0.01),and Bax and p-AKT/AKT levels decreased(P<0.01).Compared to those in the effects of miR-214-3p mimics alone,the combination of 740Y-P reversed the above effects of miR-214-3p mimics,while the combination of LY294002 further aggravated the above effects of miR-214-3p mimics.Conclusion miR-214-3p promotes OA cell proliferation and autophagy,and inhibits apoptosis,which may be related to the inhibition of the activation of PI3K/AKT pathway.

关 键 词：miR-214-3p 骨关节炎 软骨细胞 自噬 凋亡 PI3K/AKT通路 

分 类 号：R684[医药卫生—骨科学]