外源性胆绿素对氧糖缺失-复氧复糖损伤PC12细胞Litaf表达的影响  

Effects of exogenous biliverdin on expression of Litaf in PC12 cells subjected to oxygen-glucose deprivation and restoration

作　　者：李俊杰[1] 邹志瑶 彭丽佳 罗靖 熊莉 刘佳[1] 王廷华[1] 邵建林[1] Li Junjie;Zou Zhiyao;Peng Lijia;Luo Jing;Xiong Li;Liu Jia;Wang Tinghua;Shao Jianlin(Department of Anesthesiology,First Affiliated Hospital of Kunming Medical University,Kunming 650000,China)

机构地区：[1]昆明医科大学第一附属医院麻醉科,650000

出　　处：《中华麻醉学杂志》2020年第3期347-349,共3页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(81760248,81960250);云南省应用基础研究重点项目(2018FA042)。

摘　　要：目的评价外源性胆绿素对氧糖缺失-复氧复糖损伤PC12细胞Litaf表达的影响。方法将PC12细胞以1×104个/孔的密度接种于96孔细胞培养板培养3 d,采用随机数字表法分为3组(n=18):对照组(C组)、氧糖缺失-复氧复糖组(OGD/R组)和胆绿素组(BV组)。C组37℃培养箱(95%空气+5%CO2)中培养6 h;采用无糖培养基,37℃培养箱(95%N2+5%CO2)中培养2 h后将培养基更换为正常培养基,37℃培养箱(95%空气+5%CO2)继续培养的方法制备PC12细胞氧糖缺失-复氧复糖模型,BV组于复氧复糖即刻加入2μg/ml胆绿素孵育。于复氧复糖6 h时每组随机取6孔,采用Western blot和RT-PCR法检测Litaf及其mRNA表达,采用ELISA法检测上清液TNF-α浓度。结果与C组比较,OGD/R组Litaf及其mRNA表达上调,上清液TNF-α浓度升高(P<0.05);与OGD/R组比较,BV组Litaf及其mRNA表达下调,上清液TNF-α浓度降低(P<0.05)。结论外源性胆绿素减轻PC12细胞氧糖缺失-复氧复糖损伤的机制与抑制Litaf表达上调,减轻炎症反应有关。Objective To evaluate the effects of exogenous biliverdin(BV)on the expression of Litaf in PC12 cells subjected to oxygen-glucose deprivation and restoration(OGD/R).Methods PC12 cells were seeded in a 96-well cell culture plate at a density of 1×104 cells/well for 3 days and were divided into 3 groups(n=18 each)by a random number table method:control group(group C),OGD/R group,and biliverdin group(BV group).Group C was incubated in a 37℃incubator(95%air+5%CO2)for 6 h.To establish the OGD/R model,cells were incubated with sugar-free medium in a 37℃incubator(95%air+5%CO2)for 2 h,and the medium was then replaced with normal medium and cells were continuously incubated in a 37℃incubator(95%N2+5%CO2).In BV group,2μg/ml biliverdin was added immediately after oxygen-glucose restoration.Cells in 6 wells in each group were selected at 6 h of restoration for determination of the expression of Litaf protein and mRNA(by real-time polymerase chain reaction)and tumor necrosis factor-alpha(TNF-α)concentration(by enzyme-linked immunosorbent assay).Results Compared with group C,the expression of Litaf protein and mRNA was significantly up-regulated,and TNF-αconcentration in supernatant was increased in group OGD/R(P<0.05).Compared with group OGD/R,the expression of Litaf protein and mRNA was significantly down-regulated,and TNF-αconcentration in supernatant was decreased in group BV(P<0.05).Conclusion The mechanism by which exogenous biliverdin reduces OGD/R damage to PC12 cells is related to inhibiting up-regulated expression of Litaf and alleviating the inflammatory responses.

关 键 词：胆绿素 PC12细胞 低氧 Litaf 

分 类 号：R743[医药卫生—神经病学与精神病学]