α_2-肾上腺素能受体激动剂B-TH933抑制LPS处理的心肌细胞释放TNF-α  

α_2-adrenoceptor agonist B-HT933 suppresses LPS-induced TNF-α production in neonatal rat cardiomyocytes

作　　者：朱琳欣 杨多猛 唐翔诩 王媛[2] 吕秀秀[2] 李红梅[2] 严玉霞[3] 戚仁斌[2] 陆大祥[2] 王华东[2] 

机构地区：[1]暨南大学医学院临床医学系,广东广州510632 [2]暨南大学医学院病理生理学系,国家中医药管理局三级科研实验室,广东广州510632 [3]暨南大学医学院生物化学系,广东广州510632

出　　处：《中国病理生理杂志》2015年第9期1595-1600,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81170222;No.81372028);广东省教育厅学科建设科技创新项目(No.2013KJCX0019);广州市科技计划(No.12C22071599;No.201508020005)

摘　　要：目的:观察α2-肾上腺素能受体激动剂B-HT933对脂多糖(LPS)刺激的心肌细胞产生肿瘤坏死因子α(TNF-α)的影响,并初步分析其作用机制。方法:分离培养SD大鼠乳鼠心肌细胞。利用免疫荧光染色观察心肌α2A-肾上腺素能受体的分布;B-HT933和/或LPS处理心肌细胞一定时间后,用ELISA方法检测细胞培养液中TNF-α的含量、实时荧光定量PCR测定心肌细胞TLR4和TNF-αmRNA的表达、Western blot分析心肌细胞中相关信号分子的磷酸化水平。结果:免疫荧光染色证实乳鼠心肌细胞中存在α2A-肾上腺素能受体。LPS以剂量和时间依赖的方式刺激心肌细胞产生TNF-α,0.1μmol/L的B-HT933处理能显著抑制LPS诱导的TNF-αmRNA的表达和TNF-α蛋白的产生。而且,BHT能抑制LPS诱导的心肌细胞IκBα的磷酸化。结论:乳鼠心肌细胞上存在α2A-肾上腺素能受体,其激动剂B-HT933可能通过抑制心肌细胞IκBα的磷酸化来减少LPS诱导的TNF-α产生。AIM：To observe the effect of B-HT933, a selective α2-adrenoceptor agonist, on lipopolysaccha-ride （ LPS ）-induced TNF-αproduction in neonatal rat cardiomyocytes and to explore the underlying mechanisms . METHODS：The neonatal rat cardiomyocytes were cultured .The localization of α2A-adrenoceptor in the cardiomyocytes was examined by immunofluorescence staining .The cardiomyocytes were exposed to LPS or/and B-HT933 for different time.The level of TNF-αin the supernatants and the mRNA expression of TNF-αwere detected by ELISA and real-time PCR, respectively.In addition, LPS-associated signal molecules in the cardiomyocytes were also examined by Western blotting.RESULTS： Immunofluorescence staining showed that α2A-adrenoceptors were localized in the cardiomyocytes . LPS stimulated TNF-αproduction in the cardiomyocytes in a dose and time-dependent manner .B-HT933 pretreatment sig-nificantly inhibited the expression of TNF-αat mRNA and protein levels in LPS-treated cardiomyocytes .Furthermore, LPS exposure induced IκBαand p38 phosphorylation in cardiomyocytes and only IκBαphosphorylation was prevented by B-HT933 treatment.CONCLUSION：α2A-adrenoceptors are present in neonatal rat cardiomyocytes and its agonist B -HT933 inhibits LPS-induced TNF-αproduction in cardiomyocytes via suppressing IκBαphosphorylation .

关 键 词：乳鼠 心肌细胞 脂多糖 α2-肾上腺素能受体 

分 类 号：R285.5[医药卫生—中药学] R363[医药卫生—中医学]