右美托咪定调节SIRT1/PGC-1α/Nrf2信号通路对IL-1β诱导的软骨细胞损伤的影响OA
Effect of dexmedetomidine on IL-1β-induced chondrocyte injury by modulating SIRT1/PGC-1α/Nrf2 signaling pathway
目的:探究右美托咪定(DEX)调节脱乙酰酶沉默信息调节因子1(SIRT1)/PPARγ共激活因子-1α(PGC-1α)/核因子E2相关因子2(Nrf2)信号通路对IL-1β诱导的软骨细胞损伤的影响.方法:体外分离获取大鼠软骨细胞,甲苯胺蓝染色和免疫荧光用于软骨细胞鉴定.用浓度为3.125～200 nmol/L的DEX与IL-1β共同处理软骨细胞,CCK-8法检测细胞活性,筛选DEX最佳浓度;将软骨细胞分为Control组、IL-1β组、DEX低、中、高浓度组(DEX-L组、DEX-M组、DEX-H组)、DEX高浓度+SIRT1抑制剂组(DEX-H+EX527组);流式细胞术检测细胞凋亡;试剂盒测定细胞炎症因子(IL-6、TNF-α、IFN-γ)和氧化应激(ROS、MDA)水平;Western blot检测凋亡蛋白(Bax、Caspase-1)、细胞外基质相关蛋白(MMP-13、MMP3、Collagen Ⅱ)、通路蛋白表达.结果:与Control组比较,IL-1β组细胞凋亡率、Caspase-3、Bax、MMP13、MMP3表达量、IL-6、TNF-α、IFN-γ、ROS、MDA水平升高,Collagen Ⅱ、SIRT1、PGC-1α、Nrf2、Nrf1、p-AMPK/AMPK、TFAM表达量降低(P<0.05);与IL-1β组比较,DEX-L、DEX-M、DEX-H组细胞凋亡率、Caspase-3、Bax、MMP13、MMP3表达量、IL-6、TNF-α、IFN-γ、ROS、MDA水平降低,Collagen Ⅱ、SIRT1、PGC-1α、Nrf2、Nrf1、p-AMPK/AMPK、TFAM表达量升高(P<0.05);与DEX-H组相比,DEX-H+EX527组细胞凋亡率、Caspase-3、Bax、MMP13、MMP3表达量、IL-6、TNF-α、IFN-γ、ROS、MDA水平上升,Collagen Ⅱ、SIRT1、PGC-1α、Nrf2、Nrf1、p-AMPK/AMPK、TFAM表达量下降(P<0.05).结论:DEX通过激活SIRT1/PGC-1α/Nrf2信号通路缓解IL-1β诱导的软骨细胞损伤.
Objective:To investigate the effect of dexmedetomidine(DEX)on IL-1β-induced chondrocyte injury by modulating silent information regulator 1(SIRT1)/PPARγ coactivator factor-1α(PGC-1α)/nuclear factor E2 related factor 2(Nrf2)signaling pathway.Methods:Rat chondrocytes were isolated and cultured in vitro,toluidine blue staining and immunofluorescence were used to identify chondrocytes.Chondrocytes were co-treated with DEX at concentrations ranging from 3.125 to 200 nmol/L and IL-1β.CCK-8 method was used to detect cell viability and screen for the optimal concentration of DEX.Chondrocytes were assigned into Control group,IL-1β group,DEX low,medium and high concentration groups(DEX-L group,DEX-M group,DEX-H group)and DEX high concentration+SIRT1 inhibitor group(DEX-H+EX527 group).Cell apoptosis was detected by flow cytometry.Levels of cellular inflam-matory factors(IL-6,TNF-α,IFN-γ)and oxidative stress(ROS,MDA)were measured by kits.Western blot was used to measure expressions of apoptotic proteins(Bax,Caspase-1),extracellular matrix related proteins(MMP-13,MMP3,Collagen Ⅱ),and path-way proteins.Results:Compared with Control group,apoptosis rate,Caspase-3,Bax,MMP13,MMP3 expression levels,IL-6,TNF-α,IFN-γ,ROS and MDA levels were increased in IL-1β group,while Collagen Ⅱ,SIRT1,PGC-1α,Nrf2,Nrf1,p-AMPK/AMPK and TFAM expression levels were decreased(P<0.05).Compared with IL-1β group,apoptosis rate,Caspase-3,Bax,MMP13,MMP3 expression levels,IL-6,TNF-α,IFN-γ,ROS and MDA levels were decreased in DEX-L,DEX-M,and DEX-H groups,while the Collagen Ⅱ,SIRT1,PGC-1α,Nrf2,Nrf1,p-AMPK/AMPK and TFAM expression levels were increased(P<0.05).Compared with DEX-H group,apoptosis rate,Caspase-3,Bax,MMP13,MMP3 expression levels,IL-6,TNF-α,IFN-γ,ROS and MDA levels were increased in DEX-H+EX527 group,while Collagen Ⅱ,SIRT1,PGC-1α,Nrf2,Nrf1,p-AMPK/AMPK and TFAM expression levels were decreased(P<0.05).Conclusion:DEX alleviates IL-1β-induced chondrocyte injury by activating the SIRT1/PGC-1α/Nrf2 signaling pathway.
刘洪芳;杨猛;刘希明
烟台市烟台山医院药剂科,烟台 264001滕州市中心人民医院创伤骨科,滕州 277500滕州市中心人民医院麻醉科,滕州 277500
医药卫生
右美托咪定SIRT1/PGC-1α/Nrf2信号通路IL-1β诱导软骨细胞损伤
DexmedetomidineSIRT1/PGC-1α/Nrf2 signaling pathwayIL-1β inductionChondrocyte injury
《中国免疫学杂志》 2026 (8)
1834-1839,1846,7
徐州医科大学附属医院资助项目(XYFM202422).
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