布托啡诺对白细胞介素-1β诱导的关节软骨细胞焦亡的影响及机制研究  

Effect of butorphanol on interleukin-1βinduced pyroptosis of articular chondrocytes and its mechanism

作　　者：张玲 陈尧 尹航 程超 ZHANG Ling;CHEN Yao;YIN Hang;CHENG Chao(Department of Anesthesiology,Huangshi Hospital of Traditional Chinese Medicine,Huangshi 435000,China)

机构地区：[1]黄石市中医医院麻醉科,湖北黄石435000 [2]黄石市中医医院骨科,湖北黄石435000

出　　处：《陕西医学杂志》2023年第12期1665-1669,1674,共6页Shaanxi Medical Journal

基　　金：湖北省黄石市医疗卫生科技计划项目(2019A29)。

摘　　要：目的:探讨布托啡诺(BT)对白细胞介素-1β(IL-1β)诱导的关节软骨细胞焦亡的影响及相关机制。方法:取对数生长期HC-a细胞,分为对照组、IL-1β组(给予10 ng/ml IL-1β)、BT低浓度(BT-L)组(给予10 ng/ml IL-1β+1μmol/L BT)、BT中浓度(BT-M)组(给予10 ng/ml IL-1β+2μmol/L BT)、BT高浓度(BT-H)组(给予10 ng/ml IL-1β+4μmol/L BT)、BT-H+NOD样受体蛋白3(NLRP3)激活剂(ATP)组(给予10 ng/ml IL-1β+4μmol/L BT+3 mmol/L ATP)。除对照组外,其余各组均经相应浓度物质处理HC-a细胞。48 h后,倒置显微镜观察各组细胞形态变化;MTT法检测细胞增殖率;Hoechst 33342/碘化丙啶(PI)染色检测细胞焦亡变化;实时荧光定量PCR(RT-qPCR)检测细胞NLRP3、半胱天冬酶-1(Caspase-1)、IL-1βmRNA表达;ELISA法检测细胞一氧化氮(NO)、肿瘤坏死因子-α(TNF-α)及IL-6水平;Western blot检测NLRP3/Caspase-1信号通路及IL-1β表达水平。结果:与对照组比较,IL-1β组PI阳性细胞率,NO、IL-6、TNF-α水平,IL-1β、Caspase-1、NLRP3蛋白及mRNA表达增加,细胞增殖率显著下降(均P<0.05)。与IL-1β组比较,BT-L、BT-M、BT-H组PI阳性细胞率,NO、IL-6、TNF-α水平,IL-1β、Caspase-1、NLRP3蛋白及mRNA表达下降,细胞增殖率显著增加且呈剂量依赖性(均P<0.05)。与BT-H组比较,BT-H+ATP组PI阳性细胞率,NO、IL-6、TNF-α水平,IL-1β、Caspase-1、NLRP3蛋白及mRNA表达增加,细胞增殖率显著下降(均P<0.05)。结论:BT可以抑制IL-1β诱导的关节软骨细胞焦亡,其机制可能与抑制NLRP3/Caspase-1信号通路有关。Objective:To To investigate the effect of butorphanol(BT)on interleukin-1β(IL-1β)induced pyroptosis of articular chondrocytes and its related mechanism.Methods:HC-a cells in logarithmic growth phase were taken and divided into control group,IL-1βgroup(treated with 10 ng/ml IL-1β),BT-L group(treated with 10 ng/ml IL-1β+1μmol/L BT),BT-M group(treated with 10 ng/ml IL-1β+2μmol/L BT)BT),BT-H group(treated with 10 ng/ml IL-1β+4μmol/L BT),BT-H+NOD-like receptor protein 3(NLRP3)activator(ATP)group(treated with 10 ng/ml IL-1β+4μmol/L BT+3 mmol/L ATP).Except for the control group,HC-a cells in the other groups were treated with the corresponding concentrations of substances.After 48 hours,the morphological changes of cells in each group were observed by inverted microscope;MTT assay was used to detect cell proliferation rate;Hoechst 33342/PI staining was used to detect pyroptosis;the mRNA expressions of NLRP3,Caspase-1 and IL-1βwere detected by RT-qPCR;the levels of NO,TNF-αand IL-6 were detected by ELISA;Western blot was used to detect NLRP3/Caspase-1 signaling pathway and IL-1βexpression.Results:Compared with the control group,the PI-positive cell rate,the levels of NO,IL-6 and TNF-α,the protein and mRNA expressions of IL-1β,Caspase-1 and NLRP3 were increased,and the cell proliferation rate was decreased significantly in the IL-1βgroup(all P<0.05).Compared with the IL-1βgroup,the PI-positive cell rate,the levels of NO,IL-6 and TNF-α,and the protein and mRNA expressions of IL-1β,Caspase-1 and NLRP3 were significantly decreased,and the cell proliferation rate was significantly increased in a dose-dependent manner in the BT-L,BT-M and BT-H groups(all P<0.05).Compared with the BT-H group,the PI-positive cell rate,the levels of NO,IL-6 and TNF-α,the protein and mRNA expressions of IL-1β,Caspase-1 and NLRP3 were increased,and the cell proliferation rate was decreased significantly in the BT-H+ATP group(all P<0.05).Conclusion:BT can inhibit IL-1β-induced pyroptosis of articular chondrocytes,which may be

关 键 词：骨关节炎 布托啡诺 NOD样受体蛋白3 半胱天冬酶-1 白介素-1Β 细胞焦亡 

分 类 号：R684.3[医药卫生—骨科学]