左归丸调控Cx43对血清饥饿诱导MC3T3-E1细胞凋亡的保护作用  

Protective Effect of Zuogui Pill on Cx43 Regulation on Apoptosis of MC3T3-E1 Induced by Serum Starvation

作　　者：桑红灵[1] 周安方[1] 孙志博[2] 陶春晖[1] 赵敏[1] SANG Hongling1, ZHOU Anfang1, SUN Zhibo2, TAO Chunhui1, ZHAO Min(1. Chinese Medicine Clinical College, Hubei University of Chinese Medicine, Wuhan 430061, Hubei, China ; 2. Department of Orthopedics, Pu - Ai Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430034, Hubei, Chin)

机构地区：[1]湖北中医药大学中因临床学院,湖北武汉430061 [2]华中科技大学同济医学院附属普爱医院骨科,湖北武汉430034

出　　处：《中华中医药学刊》2018年第5期1071-1075,I0002,共6页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81403285);国家中医药管理局“周安方全国名老中医药专家传承工作室”建设项目([2014]20)

摘　　要：目的：论证左归丸含药血清抗MC3T3-E1细胞凋亡的机制与其调控Cx43表达及功能有关。方法：将培养细胞分为：正常A组、对照B组、左归丸含药血清C组。后两组采用血清饥饿法诱导细胞凋亡。流式细胞仪检测细胞凋亡、Western blot法检测Caspase3蛋白表达;RT-PCR、Western blot法分别检测Cx43mRNA、蛋白表达;免疫荧光染色检测细胞爬片Cx43蛋白分布及定量;放免法检测细胞上清液c AMP含量。结果：左归丸可明显降低血清饥饿诱导的细胞凋亡、减少caspase3蛋白表达,B、C组比较P〈0.01或P〈0.05。血清饥饿还导致Cx43mRNA及蛋白表达、蛋白荧光分布及MOD值、细胞上清液c AMP含量显著减少,而左归丸可显著提高上述指标,B、C组比较P〈0.05。结论：左归丸含药血清通过调控Cx43表达及功能,发挥抗MC3T3-E1细胞凋亡的保护作用。Objective：To demonstrate the mechanism of Zuogui Pill medicated serum anti MC3T3-E1 cells apoptosis is related to its regulation of Cx43 expression and function. Methods：The cells were divided into normal group A,control B and serum containing Zuogui Pill group C. In the latter two groups,apoptosis was induced by serum starvation. Cell apoptosis was detected by flow cytometry,and the expression of Caspase3 protein was detected at the same time. RT-PCR and Western blot were used to detect the expressions of Cx43 mRNA and protein,and then we tested the distribution and quantification of Cx43 Protein by immunofluorescence. At last,the content of c AMP was detected by radioimmunoassay.Results： Zuogui Pill could obviously reduce the apoptosis induced by serum starvation and decrease the expression of Caspase3 protein. Compared with B and C group,there was significant difference,P〈0. 01 or P〈0. 05. Serum starvation also led to the decrease of the expression of Cx43 mRNA and protein,cell protein fluorescence distribution and MOD value and the c AMP contents,while Zuogui Pill could significantly improve the above indices. Compared with B and C group,there was signficant difference,P〈0. 05. Conclusion： Zuogui Pill medicated serum can regulate the expression and function of Cx43,playing a protective role against apoptosis in MC3T3-E1 cells.

关 键 词：左归丸 含药血清 MC3T3-E1 CX43 凋亡 血清饥饿 

分 类 号：R285.5[医药卫生—中药学]