罗哌卡因诱导骨肉瘤细胞MG-63自噬性死亡和促炎因子表达  

Ropivacaine inducing autophagic death of osteosarcoma cells MG-63 and expressions of pro-inflammatory factors

作　　者：刘芳坊[1] 徐国亭[1] 刘彦锋[1] 徐国兴[1] 李海中[1] LIU Fangfang;XU Guoting;LIU Yanfeng;XU Guoxing;LI Haizhong(Department of Anesthesiology and Periopera-tive Medicine,the First Affiliated Hospital of Nanyang Medical College,Nanyang 473058,China)

机构地区：[1]南阳医学高等专科学校第一附属医院麻醉与围手术医学科,南阳473058

出　　处：《中国免疫学杂志》2022年第17期2064-2068,共5页Chinese Journal of Immunology

基　　金：2018年河南省医学科技攻关计划项目(2018020546)。

摘　　要：目的:观察罗哌卡因(Rop)对骨肉瘤MG-63细胞自噬性死亡及炎症因子表达的作用。方法:体外采用不同浓度(0、0.3、0.6、0.125、0.25、0.5、1.0、2.0、4.0、8.0、16.0 mmol/L)Rop处理骨肉瘤MG-63细胞,CCK-8筛选出0、0.5、1.0、2.0 mmol/L4个浓度,克隆形成实验检测Rop对MG-63细胞增殖的影响,Annexin V-FITC/PI双染流式细胞术检测细胞凋亡情况;荧光显微镜观察Rop对MG-63细胞自噬的影响,Western blot检测LC3Ⅱ/LC3Ⅰ、Beclin1蛋白表达;ELISA、RT-PCR分别检测Rop对MG-63细胞IL-6、iNOS分泌及IL-6、iNOS mRNA表达的影响。结果:CCK-8结果显示,Rop以剂量依赖方式抑制MG-63细胞增殖,当Rop≥1 mmol/L时,其对骨肉瘤MG-63细胞增殖具有明显抑制作用(P<0.05)。当Rop浓度为1.0、2.0 mmol/L时,与未处理组相比,MG-63细胞克隆形成率、凋亡率明显降低(P<0.05),LC3Ⅱ/LC3Ⅰ、Beclin1蛋白表达明显升高(P<0.05),且细胞质内荧光颗粒明显增多,细胞自噬率显著升高(P<0.05),IL-6、iNOS mRNA表达及IL-6、iNOS含量明显降低(P<0.05)。结论:Rop对骨肉瘤MG-63细胞具有增殖抑制作用,并能诱导MG-63细胞自噬性死亡,降低促炎因子表达。Objective:To observe effects of ropivacaine(Rop)on autophagic death of osteosarcoma cells MG-63 and expressions of inflammatory factors.Methods:Osteosarcoma MG-63 cells were treated with different concentrations of Rop(0,0.3,0.6,0.125,0.25,0.5,1.0,2.0,4.0,8.0,16.0 mmol/L)in vitro,and 4 concentrations(0,0.5,1.0,2.0 mmol/L)were screened out by CCK-8. Effect of Rop on proliferation of MG-63 cells was detected by clone formation assay. Apoptosis was detected by Annexin VFITC/PI double-staining flow cytometry. Effect of Rop on autophagy of MG-63 cells was observed by fluorescence microscope. Expressions of LC3Ⅱ/LC3Ⅰ and Beclin1 proteins were detected by Western blot. Effects of Rop on secretions of IL-6,iNOS and expressions of IL-6 and iNOS mRNA were detected by ELISA and RT-PCR.Results:CCK-8 results showed that Rop inhibited proliferation of MG-63 cells in way of dose-dependence,who showed significant inhibitory effects at Rop≥1 mmol/L(P<0.05). When Rop concentrations were 1.0 and 2.0 mmol/L,compared with non-treatment group,clone formation rate and apoptosis rate of MG-63 cells were significantly decreased(P<0.05),expressions of LC3Ⅱ/LC3Ⅰ and Beclin1 proteins were significantly increased(P<0.05),fluorescent particles in cytoplasm were significantly increased,autophagy rate was significantly increased(P<0.05),expressions of IL-6 and iNOS mRNA,contents of IL-6 and iNOS were significantly decreased(P<0.05).Conclusion:Rop can inhibit proliferation of osteosarcoma MG-63 cells,induce autophagic death,and reduce expressions of pro-inflammatory factors.

关 键 词：罗哌卡因 骨肉瘤细胞 凋亡 自噬 促炎因子 

分 类 号：R966[医药卫生—药理学]