摘要
目的探讨丙泊酚对糖氧剥夺再灌注诱导的星形胶质细胞损伤的影响及其机制。方法将体外培养的大鼠脑皮质星形胶质细胞分为对照组、模型组、模型+丙泊酚组、模型+丙泊酚+空载体组和模型+丙泊酚+高迁移率族蛋白1(HMGB1)组,后4组构建糖氧剥夺再灌注细胞损伤模型,后3组给予浓度为10μmol/L的丙泊酚处理,后两组分别在转染pcDNA3.1空载体质粒和pcDNA3.1-HMGB1过表达载体质粒后建模。采用噻唑蓝(MTT)法检测细胞存活率,比色法检测乳酸脱氢酶(LDH)漏出率,流式细胞仪检测细胞凋亡率,ELISA法检测细胞上清液中HMGB1、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)含量,Western blot检测HMGB1、B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)和活化的含半胱氨酸的天冬氨酸蛋白水解酶-3(Cleaved caspase-3)等蛋白表达。结果与对照组相比,模型组细胞存活率和Bcl-2蛋白表达水平显著降低,而LDH漏出率、凋亡率和HMGB1、Bax、Cleaved caspase-3蛋白的表达水平以及细胞上清液中HMGB1、IL-1β、TNF-α含量均显著升高(F=171.199~391.008,q=24.398~49.269,P<0.05)。给予丙泊酚处理后糖氧剥夺再灌注引起的上述变化均显著减弱(q=11.711~35.032,P<0.05)。成功上调HMGB1表达后,丙泊酚对糖氧剥夺再灌注损伤的保护作用得以逆转。结论丙泊酚可能通过下调HMGB1表达抑制细胞凋亡和炎症反应对糖氧剥夺再灌注诱导的星形胶质细胞发挥保护作用。
Objective To investigate the effect of propofol on astrocyte injury induced by glucose-oxygen deprivation and reperfusion and its mechanism.Methods Rat cerebral cortical astrocytes cultured in vitro were divided into control group,model group,model+propofol group,model+propofol+empty vector group,and model+propofol+high-mobility group box 1(HMGB1)group.The latter four groups were used to establish a model of cell injury caused by glucose-oxygen deprivation and reperfusion;the latter three groups were treated with propofol at a concentration of 10μmol/L;the latter two groups were used for modeling after being transfected with pcDNA3.1 empty vector plasmid and pcDNA3.1-HMGB1 overexpression vector plasmid,respectively.MTT assay was used to measure cell viability;colorimetry was used to measure lactate dehydrogenase(LDH)leakage rate;flow cytometry was used to measure cell apoptosis rate;ELISA was used to measure the content of HMGB1,interleukin-1β(IL-1β),and tumor necrosis factor-α(TNF-α)in cell supernatant,and Western blot was used to measure the protein expression of HMGB1,B-cell lymphoma/leukemia-2(Bcl-2),Bcl-2-associated X protein(Bax),and activated Cleaved caspase-3.ResultsCompared with the control group,the model group had significant reductions in cell viability and the protein expression level of Bcl-2 and significant increases in LDH leakage rate,cell apoptosis rate,the protein expression levels of HMGB1,Bax,and Cleaved caspase-3,and the content of HMGB1,IL-1β,and TNF-αin supernatant(F=171.199-391.008,q=24.398-49.269,all P<0.05).The above changes induced by glucose-oxygen deprivation and reperfusion were significantly alleviated after propofol treatment(q=11.711-35.032,all P<0.05).After the expression of HMGB1 was upregulated successfully,the protective effect of propofol against the injury induced by glucose-oxygen deprivation and reperfusion was reversed.Conclusion Propofol can protect astrocytes against the injury induced by glucose-oxygen deprivation and reperfusion by downregulating the expression of HMGB1 to inhibit cell apoptosis and inflammatory response.
作者
宋海军
杨族悌
刘军
刘国磊
SONG Haijun;YANG Zuti;LIU Jun;LIU Guolei(Department of Anesthesiology, Nanyang Nanshi Hospital, Nanyang 473000, China)
出处
《青岛大学学报(医学版)》
2021年第6期903-907,共5页
Journal of Qingdao University(Medical Sciences)
基金
河南省医学科技攻关计划项目(201806281)。