基于Akt/ERK探讨丹蒌片对主动脉平滑肌细胞增殖、迁移和凋亡的影响  

To Explore the Effect of Danlou Tablets on Proliferation,Migration and Apoptosis of Aortic Smooth Muscle Cells Based on Akt/ERK Signaling Pathways

作　　者：司春婴 王贺[2] 孙文博[2] 刘俊呈[1] 陈玉善 解金红[2] 关怀敏[1] Si Chunying;Wang He;Sun Wenbo;Liu Juncheng;Chen Yushan;Xie Jinhong;Guan Huaimin(The First Clinical Medical College,Henan University of Chinese Medicine,Zhengzhou 450000,China;Heart Center of The First Affiliated Hospital,Henan University of Chinese Medicine,Zhengzhou 450000,China)

机构地区：[1]河南中医药大学第一临床医学院,郑州450000 [2]河南中医药大学第一附属医院心脏中心,郑州450000

出　　处：《世界科学技术-中医药现代化》2020年第6期1883-1890,共8页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：国家自然科学基金委员会面上项目(81273949):丹蒌片促进兔髂动脉粥样硬化药物涂层支架置入后在内皮化的调控机制研究,负责人:关怀敏;国家自然科学基金委员会面上项目(81473507):化痰活血解毒方通过MAPKs信号通路抑制支架内新生动脉粥样硬化形成的机制研究,负责人:关怀敏;河南省科技攻关项目(182102310093):小檗碱联合姜黄素对ACS患者单核-巨噬细胞极化能力及分型的影响,负责人:陈玉善。

摘　　要：目的观察丹蒌片对同型半胱氨酸(homocysteine,Hcy)诱导的兔主动脉血管平滑肌细胞(rabbit aortic smooth muscle cell,RASMC)增殖、迁移和凋亡的影响,并探讨其与Akt及ERK信号通路的关系。方法培养RASMC,建立Hcy诱导RASMC增殖模型,实验分为空白组、Hcy组、丹蒌片组、瑞舒伐他汀组(即可定组)、丹蒌片联合瑞舒伐他汀组(即混合组),后三组分别给予丹蒌片、瑞舒伐他汀及丹蒌片联合瑞舒伐他汀含药血清。作用24 h后,Cell Titer-Glo法测细胞增殖,Transwell法测细胞迁移,流式细胞仪Annexin V/PI双染法测细胞凋亡率,蛋白印迹法(Western blot)测信号通路相关蛋白Akt和ERK总蛋白及磷酸化蛋白表达量。结果①与空白组相比,Hcy组细胞增殖和迁移显著增加(P<0.01),凋亡率显著降低(P<0.01);与Hcy组相比,三种含药血清组细胞增殖和迁移明显减少(P<0.01),凋亡率明显增加(P<0.01);其中混合组增殖和迁移较丹蒌片组和可定组明显降低(P<0.01),凋亡率更高(P<0.01);与可定组比,丹蒌片组增殖略高,但差异均无统计学意义(P>0.05),丹蒌片组细胞迁移数明显多于可定组(P<0.05),凋亡率明显低于可定组(P<0.05)。②各组Akt及ERK总蛋白表达水平无明显变化(P>0.05);与空白组比较,Hcy组p-Akt及p-ERK蛋白表达量均显著上调(P<0.01);与Hcy组相比,三种含药血清组p-Akt及p-ERK蛋白表达量明显下调(P<0.01);其中混合组的p-Akt及p-ERK蛋白表达较丹蒌片组和可定组进一步下降(P<0.01);与可定组比,丹蒌片组p-Akt及p-ERK蛋白表达量增加,但差异无统计学意义(P>0.05)。结论丹蒌片可拮抗Hcy诱导的兔动脉血管平滑肌细胞增殖和迁移,促进其凋亡,其作用可能与上调p-Akt及p-ERK信号通路蛋白有关。Objective To observe the effects of Danlou Tablets on the proliferation,migration and apoptosis of rabbit aortic vascular smooth muscle cells(RASMC)induced by homocysteine(Hcy),and to explore its relationship with Akt and ERK signaling pathways.Methods The model of proliferation of rabbit aortic vascular smooth muscle cells was established by Hcy.After 24 hours of different drug-containing serum,Cell Titer-Glo method was used to measure cell proliferation,transwell method was used to measure cell migration,flow cytometry Annexin V/PI double staining method was used to detect cell apoptosis rate,western blot was used to analysis protein expression of the total and phosphorylated Akt and ERK protein.Results①Compared with the control group,the proliferation and migration of Hcy group were significantly increased(P<0.01),and the apoptosis rate was significantly decreased(P<0.01).Compared with Hcy group,the proliferation,migration and apoptosis rate of three drug-containing serum groups were obvious decreased(P<0.01).However,the proliferation and migration of Danlou tablets group and Rosuvastatin group were significantly higher than those of the mixed group(P<0.01),and the apoptosis rate was significantly lower than that of the mixed group(P<0.01).Compared with the Rosuvastatin group,the proliferation of Danlou tablets was slightly higher,but the difference was not statistically significant(P>0.05).The cells migration numbers of Danlou tablets group were significantly higher than that of the determinable group(P<0.05).The apoptosis rate was significantly lower than the determinable group(P<0.05).②There was no significant change in the expression of total protein of Akt and ERK in each group(P>0.05).Compared with the control group,the expression of p-Akt and p-ERK protein in Hcy group were significantly up-regulated(P<0.01);Compared with Hcy group,the protein expression levels of p-Akt and pERK in three drug-containing serum groups were significantly down-regulated(P<0.01).However,compared with the mixed group,the

关 键 词：血管平滑肌细胞 增殖 迁移 凋亡 支架内再狭窄 丹蒌片 

分 类 号：R282[医药卫生—中药学]