神经节苷脂预处理对罗哌卡因诱导SH-SY5Y细胞焦亡的神经保护作用  

Neuroprotective effect of ganglioside pretreatment on pyroptosis of SH-SY5Y cells induced by ropivacaine

作　　者：刘丹[1] 刘敬臣[1] Liu Dan;Liu Jingchen(Department of Anesthesiology,The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院麻醉科,南宁530021

出　　处：《广西医科大学学报》2021年第2期236-242,共7页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81660623);广西研究生教育创新计划项目资助(No.YCSW2020117)。

摘　　要：目的:探讨神经节苷脂(GM-1)抑制罗哌卡因诱导神经母细胞瘤细胞(SH-SY5Y)发生焦亡的神经保护作用及分子机制。方法:将对数生长期SH-SY5Y细胞随机分为对照组(C组):无罗哌卡因或GM-1干预;GM-1预处理组(G组):SH-SY5Y细胞中加入5μmol/L的GM-1培养24 h;GM-1预处理组(G+R组):GM-1预处理24 h后,SH-SY5Y细胞中加入1.5 mmol/L的罗哌卡因培养24 h;罗哌卡因组(R组):SH-SY5Y细胞中加入1.5 mmol/L的罗哌卡因培养24 h。分别通过MTT法测定细胞活力、TUNEL染色法检测细胞凋亡、酶联免疫吸附测定法检测细胞上清炎症因子白细胞介素(IL)-1β、IL-18含量、免疫荧光法和蛋白印迹法检测焦亡相关因子Gasdermin D(GSDMD)、Nod样受体蛋白3(NLRP3)、半胱氨酸天冬氨酸酶-1(caspase-1)表达差异。结果:与C组比较,G+R组和R组SH-SY5Y细胞存活率明显降低(P<0.05),细胞凋亡率、IL-1β、IL-18含量及焦亡相关蛋白(GSDMD、NLRP3、caspase-1)表达水平升高(P<0.05),G组细胞凋亡率显著增加(P<0.05),其他指标无显著变化(P>0.05);与G组相比,G+R组和R组上述各指标均有统计学差异(P<0.05);与R组相比,G+R组上述指标变化均显著改善(P<0.05)。结论:GM-1可通过减少GSDMD、NLRP3和caspase-1表达从而减轻焦亡,有效抑制罗哌卡因所致SH-SY5Y细胞的神经毒性作用。Objective: To explore the neuroprotective effect and the molecular mechanism of ganglioside(GM-1) pretreatment on ropivacaine-induced pyroptosis in neuroblastoma cells(SH-SY5 Y). Methods: SH-SY5 Y cells were randomly divided into four groups: control group(C group): without any intervention of ropivacaine or GM-1;GM-1 pretreatment group(G group): SH-SY5 Y cells were cultured with 5 μmol/L GM-1 for 24 h;GM-1 pretreatment group(G+R group): After GM-1 pretreatment for 24 h, SH-SY5 Y cells were cultured with 1.5 mmol/L ropivacaine for 24 h;Ropivacaine group(R group): SH-SY5 Y cells were cultured with 1.5 mmol/L ropivacaine for 24 h. Cell viability was measured by MTT method. Cell apoptosis was detected by TUNEL staining method.Inflammatory factors IL-1β and IL-18 contents in cell supernatant were detected by enzyme-linked immunosorbent assay. Immunofluorescence method and Western blotting were used to detect the expressions of pyroptosisrelated factor Gasdermin D(GSDMD), NOD like receptor thermoprotein domain 3(NLRP3) and cysteine-proteinase-1(caspase-1). Results: Compared with the C group, the survival rate of SH-SY5 Y cells was significantly decreased in the G+R group and R group(P<0.05), and the rate of apoptosis, the contents of IL-1β and IL-18,and the protein expressions of GSDMD, NLRP3 and caspase-1 were significantly increased(P<0.05). The apoptosis rate was markedly increased in the G group(P<0.05), while the other indicators showed no statistic difference(P>0.05). Compared with the G group, the above-mentioned indicators showed statistical significance in the G+R group and R group(P<0.05).Compared with the R group, the changes of the above-mentioned indicators in the G+R group were significantly improved(P<0.05). Conclusion: GM-1 can reduce pyroptosis by reducing the expression of GSDMD, NLRP3 and caspase-1, and effectively inhibit the neurotoxic effect of ropivacaine on SH-SY5 Y cells.

关 键 词：神经节苷脂(GM-1) 罗哌卡因 SH-SY5Y细胞 焦亡 

分 类 号：R96[医药卫生—药理学]