蓝萼甲素调节PINK1/Parkin信号通路对IL-1β诱导的关节软骨细胞自噬和凋亡的影响  

Effects of GLA on IL-1β-Induced Autophagy and Apoptosis in Articular Chondrocytes via PINK1/Parkin Pathway

作　　者：高翔 李巍 陈何 GAO Xiang;LI Wei;CHEN He(Chengdu Bayi Orthopedic Hospital,Sichuan Chengdu 610091,China)

机构地区：[1]四川省成都市八一骨科医院骨科,四川成都610091

出　　处：《河北医学》2024年第8期1244-1249,共6页Hebei Medicine

基　　金：四川省卫生健康委员会科研课题,(编号:19PJ100)。

摘　　要：目的:探讨蓝萼甲素(GLA)调节PTEN诱导的激酶1(PINK1)/E3泛素连接酶(Parkin)信号通路对白细胞介素1β(IL-1β)诱导的关节软骨细胞自噬和凋亡的影响。方法:将关节软骨细胞分为Control组、IL-1β组、L-GLA组、M-GLA组、H-GLA组、Suramin组;细胞计数试剂盒8(CCK8)法检测关节软骨细胞增殖;TEM检测关节软骨细胞自噬;流式细胞仪检测关节软骨细胞凋亡;ELISA检测关节软骨细胞中炎性因子(MMP3、TNF-α、MMP13)和活性氧(ROS)水平;蛋白印迹法检测关节软骨细胞中裂解的胱天蛋白酶3(cleaved caspase-3)、微管相关蛋白1轻链3(LC3)I、LC3Ⅱ、P62、PINK1、Parkin蛋白表达。结果:与Control组相比,IL-1β组存活率、LC3Ⅱ/Ⅰ、PINK1、Parkin降低,自噬空泡数、凋亡率、MMP3、TNF-α、MMP13、ROS、cleaved caspase-3、P62升高(P<0.05);L-GLA组、M-GLA组、H-GLA组存活率、自噬空泡数、LC3Ⅱ/Ⅰ、PINK1、Parkin高于IL-1β组,凋亡率、MMP3、TNF-α、MMP13、ROS、cleaved caspase-3、P62低于IL-1β组(P<0.05);Suramin组存活率、自噬空泡数、LC3Ⅱ/Ⅰ、PINK1、Par-kin低于H-GLA组,凋亡率、MMP3、TNF-α、MMP13、ROS、cleaved caspase-3、P62高于H-GLA组(P<0.05)。结论:GLA通过激活IL-1β诱导的关节软骨细胞自噬,抑制细胞凋亡,进而抑制OA的进展,其机制可能是通过激活PINK1/Parkin信号通路实现的。Objective:To investigate the effect of geniposide(GLA)on PTEN-induced kinase 1(PINK1)/E3 ubiquitin ligase(Parkin)signaling pathway in regulating autophagy and apoptosis of articular chondrocytes induced by interleukin-1β(IL-1β).Methods:Articular chondrocytes were divided into Control,IL-1β,Low-GLA,Medium-GLA,High-GLA,and Suramin groups.Cell proliferation was assessed using the Cell Counting Kit-8(CCK8)assay.Autophagy was detected by transmission electron microscopy(TEM).Apoptosis was measured by flow cytometry.Levels of inflammatory factors(MMP3,TNF-α,MMP13)and reactive oxygen species(ROS)in chondrocytes were detected by ELISA.Protein expressions of cleaved caspase-3,microtubule-associated protein 1 light chain 3(LC3)I,LC3II,P62,PINK1,and Parkin were analyzed by Western blot.Results:Compared with the Control group,the IL-1βgroup showed decreased cell viability,LC3II/I ratio,PINK1,and Parkin,while autophagic vacuole number,apoptosis rate,MMP3,TNF-α,MMP13,ROS,cleaved caspase-3,and P62 were increased(P<0.05).The Low-GLA,Medium-GLA,and High-GLA groups exhibited higher cell viability,autophagic vacuole number,LC3II/I ratio,PINK1,and Parkin than the IL-1βgroup,with lower apoptosis rate,MMP3,TNF-α,MMP13,ROS,cleaved caspase-3,and P62(P<0.05).The Suramin group showed lower cell viability,autophagic vacuole number,LC3II/I ratio,PINK1,and Parkin than the High-GLA group,with higher apoptosis rate,MMP3,TNF-α,MMP13,ROS,cleaved caspase-3,and P62(P<0.05).Conclusion:GLA inhibits the progression of osteoarthritis(OA)by activating autophagy and inhibiting apoptosis in IL-1β-induced articular chondrocytes,possibly through the activation of the PINK1/Parkin signaling pathway.

关 键 词：蓝萼甲素 PTEN诱导的激酶1 E3泛素连接酶 IL-1Β 关节软骨细胞 自噬 凋亡 

分 类 号：R684.3[医药卫生—骨科学]