罗哌卡因调节沉默信息调控基因1/叉头转录因子1信号通路对白细胞介素-1β诱导的软骨细胞损伤的影响实验研究  

Effect of ropivacaine on interleukin-1β-induced chondrocyte injury by regulating the SIRT1/FOXO1 signaling pathway

作　　者：王海龙[1] 夏丰娜 高学锋 王朝阳 唐楠[1] 马洲佩 WANG Hailong;XIA Fengna;GAO Xuefeng;WANG Chaoyang;TANG Nan;MA Zhoupei(Department of Anesthesiology,the Seventh People’s Hospital of Hebei,Dingzhou 073000,China;Department of Orthopedics,the Seventh People’s Hospital of Hebei,Dingzhou 073000,China)

机构地区：[1]河北省第七人民医院河北中医药大学第二附属医院麻醉科,河北定州073000 [2]河北省第七人民医院河北中医药大学第二附属医院骨科,河北定州073000

出　　处：《陕西医学杂志》2025年第3期314-318,332,共6页Shaanxi Medical Journal

基　　金：河北省医学科学研究课题(20190781)。

摘　　要：目的:探究罗哌卡因(ROP)对白细胞介素(IL)-1β诱导的软骨细胞损伤的影响及其作用机制。方法:将原代大鼠软骨细胞分为对照(NC)组、模型组、罗哌卡因低剂量(ROP-L)组、罗哌卡因高剂量(ROP-H)组和ROP-H+EX-527组。除NC组外,其余各组均加入IL-1β(10 ng/ml)处理,以构建OA软骨细胞损伤模型。24 h后,ROP-L组和ROP-H组分别加入25、100 mg/L ROP,ROP-H+EX-527组加入100 mg/L ROP和10μmol/L沉默信息调控基因1(SIRT1)抑制剂EX-527培养。实时荧光定量PCR(RT-qPCR)检测软骨细胞中SIRT1、叉头转录因子1(FOXO1)mRNA水平;CCK-8法检测各组软骨细胞增殖情况;TUNEL法检测各组软骨细胞凋亡情况;ELISA检测软骨细胞中IL-8、肿瘤坏死因子-α(TNF-α)、IL-6水平;Western blot检测软骨细胞SIRT1/FOXO1信号通路及基质降解相关蛋白表达水平。结果:与NC组比较,模型组软骨细胞增殖率、SIRT1 mRNA及蛋白水平、FOXO1 mRNA及p-FOXO1/FOXO1水平、Ⅱ型胶原α1(COL2A1)、软骨蛋白聚糖(ACAN)蛋白表达水平降低,细胞凋亡率、IL-8、TNF-α、IL-6、基质金属蛋白酶-13(MMP-13)蛋白表达水平升高(均P<0.05)。与模型组比较,ROP-L和ROP-H组软骨细胞增殖率、SIRT1 mRNA及蛋白水平、FOXO1 mRNA及p-FOXO1/FOXO1水平、COL2A1、ACAN蛋白表达水平升高,细胞凋亡率、IL-8、TNF-α、IL-6水平及MMP-13蛋白表达降低,且作用随着ROP剂量的增加而增强(均P<0.05)。EX-527逆转了ROP对OA软骨细胞的作用。结论:ROP通过激活SIRT1/FOXO1信号通路,增加软骨细胞增殖率,减少细胞凋亡,缓解IL-1β诱导的软骨细胞损伤。Objective:To investigate the effect and mechanism of ropivacaine(ROP)on interleukin-1β(IL-1β)-induced chondrocyte injury.Methods:The primary rat chondrocytes were divided into control(NC)group,model group,Ropivacaine low-dose(ROP-L)group,Ropivacaine high-dose(ROP-H)group and ROP-H+EX-527 group.Except the NC group,the other groups were treated with IL-1β(10 ng/ml)to construct the OA chondrocyte injury model.After 24 hours,the ROP-L group and the ROP-H group were added with 25 mg/L ROP and 100 mg/L ROP,respectively,and the ROP-H+EX-527 group was added with 100 mg/L ROP and 10μmol/L inhibitor of silenced information regulatory gene 1(SIRT1)EX-527.RT-qPCRwas applied to detect the mRNA levels of SIRT1 and FOXO1 in chondrocytes.Chondrocyte proliferation was detected by CCK-8 method.Chondrocyte apoptosis was detected by TUNEL method.ELISA was applied to detect the levels of IL-8,TNF-α,and IL-6 in chondrocytes.Western blot was performed to detect SIRT1/FOXO1 signaling pathway and matrix degradation related protein expression levels in chondrocytes.Results:Compared with the NC group,the proliferation rate of chondrocytes,SIRT1 mRNA and protein levels,FOXO1 mRNA and p-FOXO1/FOXO1 levels,COL2A1 and ACAN protein expression levels in the model group were obviously lower,while the apoptosis rate,IL-8,TNF-α,IL-6,and MMP-13 protein expression levels were obviously higher(all P<0.05).Compared with the model group,the proliferation rate of chondrocytes,SIRT1 mRNA and protein levels,FOXO1 mRNA and p-FOXO1/FOXO1 levels,COL2A1 and ACAN protein expression levels in ROP-L group and ROP-H group were obviously higher,while the apoptosis rate,IL-8,TNF-α,IL-6,and MMP-13 protein expression levels were obviously lower,and the effect enhanced with the increase of ROP dose(all P<0.05).SIRT1 inhibitor EX-527 reversed the effect of ROP on OA chondrocytes.Conclusion:ROP can increase the proliferation rate of chondrocytes,reduce cell apoptosis and alleviate IL-1β-induced chondrocyte injury by activating SIRT1/FOXO1 signaling pathway.

关 键 词：软骨细胞损伤 罗哌卡因 沉默信息调控基因1 叉头转录因子1 白细胞介素-1Β 细胞模型 

分 类 号：R691.3[医药卫生—泌尿科学]