程序化细胞死亡因子5对肿瘤坏死因子-α诱导大鼠关节软骨细胞损伤的影响  

Effect of PDCD5 on articular chondrocyte injury induced by TNF-α

作　　者：华植 韦嵩 陈志煌 李晓昊 张娴娴 侯春福 刘颖琬 李慧 HUA Zhi;WEI Song;CHEN Zhi-Huang(Traditional Chinese Medicine Hospital of Guangzhou General Hospital of PLA, Guangzhou 510010, Guangdong, China)

机构地区：[1]解放军南部战区总医院中医科,广东广州510010 [2]广州中医药大学

出　　处：《中国老年学杂志》2019年第18期4519-4523,共5页Chinese Journal of Gerontology

基　　金：国家自然科学基金(No.81573883)

摘　　要：目的研究程序化细胞死亡因子(PDCD)5对肿瘤坏死因子(TNF)-α诱导的大鼠关节软骨细胞损伤的影响。方法以实时聚合酶链反应(PCR)和Western印迹分别测定关节软骨细胞经TNF-α诱导后细胞中PDCD5表达水平。用PDCD5 siRNA慢病毒感染关节软骨细胞,检测细胞中PDCD5表达水平。噻唑蓝(MTT)法测定细胞增殖,流式细胞术测定细胞凋亡,Western印迹检测细胞中剪切型Caspase(Cleaved Caspase)-3、-9蛋白水平,试剂盒检测细胞中超氧化物歧化酶(SOD)和一氧化氮合酶(NOS)活性。结果 TNF-α诱导后的关节软骨细胞中PDCD5 mRNA和蛋白水平升高。PDCD5 siRNA慢病毒感染能够下调TNF-α条件下关节软骨细胞中PDCD5表达。关节软骨细胞经TNF-α诱导以后,细胞增殖能力降低,细胞凋亡率升高,细胞中Cleaved Caspase-3、-9蛋白水平升高,细胞中SOD活性降低,NOS活性升高,与未经TNF-α诱导的细胞比较,差异有统计学意义(P<0.05)。沉默PDCD5后的关节软骨细胞经TNF-α诱导以后,细胞的增殖能力升高,细胞凋亡率降低,细胞中Cleaved Caspase-3、-9蛋白水平降低,细胞中SOD活性升高,NOS活性降低,与没有沉默PDCD5的细胞比较,差异有统计学意义(P<0.05)。结论沉默PDCD5能够减少TNF-α诱导的关节软骨细胞凋亡,减少细胞损伤。Objective To study the effect of PDCD5 on articular chondrocyte injury induced by TNF-α.Methods The expression level of PDCD5 in articular chondrocytes was determined by Realtime PCR and Western blot. PDCD5 siRNA lentivirus was used to infect articular cartilage cells, then the expression of PDCD5 in the cells was detected. MTT assay was used to determine cell proliferation. Cell apoptosis was measured by flow cytometry. Western blot was used to detect the levels of Cleaved Caspase-3 and Cleaved Caspase-9 protein in the cells. The activities of SOD and NOS in the cells were detected by kit. Results PDCD5 mRNA and protein levels in articular chondrocytes were increased after TNF-α induction. PDCD5 siRNA lentivirus infection silenced PDCD5 expression in articular chondrocytes under TNF-α condition. After induced by TNF-α, the proliferation of cells were reduced, the rate of apoptosis was increased, the levels of Cleaved Caspase-3 and Cleaved Caspase-9 protein were increased, the activity of SOD was reduced, the activity of NOS was elevated, the differences were statistically significant(P<0.05). After silencing PDCD5, the articular cartilage cells were induced, the proliferation of cells were increased, the rate of apoptosis was reduced, the levels of Cleaved Caspase-3 and Cleaved Caspase-9 protein were decreased, the activity of SOD was increased, the activity of NOS was reduced, the differences were statistically significant(P<0.05).Conclusions Silencing PDCD5 could reduce the apoptosis,reduce cell damage of articular chondrocytes induced by TNF-α.

关 键 词：关节软骨细胞 肿瘤坏死因子(TNF)-α 程序化细胞死亡因子(PDCD)5 凋亡 

分 类 号：R684.3[医药卫生—骨科学]