芪白平肺胶囊通过抑制PI3K/Akt通路促进低氧大鼠肺动脉平滑肌细胞凋亡  

Qibai Pingfei Capsule Promotes the Apoptosis of Pulmonary Artery Smooth Muscle Cells in Rats with Hypoxia by Inhibiting the Phosphoinositide 3-Kinase/Protein Kinase B Pathway

作　　者：张璐 童祥丽 王小乐[1] 朱洁[1] 李泽庚[1,2,3] ZHANG Lu;TONG Xiang-li;WANG Xiao-le;ZHU Jie;LI Ze-geng(Anhui University of Chinese Medicine, Anhui Hefei 230012, China;The First Affiliated Hospital of Anhui University of Chinese Medicine, Anhui Hefei 230031, China;Institute of Respiratory Disease Prevention and Treatment of Traditional Chinese Medicine, Anhui Academy of Chinese Medicine, Anhui Hefei 230031, China)

机构地区：[1]安徽中医药大学,安徽合肥230012 [2]安徽中医药大学第一附属医院,安徽合肥230031 [3]安徽省中医药科学院中医呼吸病防治研究所,安徽合肥230031

出　　处：《安徽中医药大学学报》2022年第2期55-60,共6页Journal of Anhui University of Chinese Medicine

基　　金：国家自然科学基金项目(U20A20398,81974569);安徽省自然科学基金青年项目(1908085QH369)。

摘　　要：目的从细胞水平探讨芪白平肺胶囊通过磷脂酰肌醇3激酶(phosphatidylinositol 3-kinasePI3K/蛋白激酶B protein kinas B,Akt)通路对低氧诱导的大鼠肺动脉平滑肌细胞(pulmonary arterial smooth muscle cells,PASMCs)凋亡的影响。方法采用组织贴块法分离培养大鼠PASMCs,制备含药血清作用于低氧诱导的大鼠PASMCs,将细胞分为常氧组、低氧组、低氧+芪白平肺胶囊含药血清组、低氧+芪白平肺胶囊含药血清组+LY294002组。采用细胞计数试剂盒检测芪白平肺胶囊含药血清对低氧诱导的大鼠PASMCs活性的影响;蛋白免疫印迹法检测芪白平肺胶囊含药血清单独及加入LY294002对Akt、磷酸化Akt(phosphorylated-Akt,p-Akt)、B淋巴细胞瘤2(B cell lymphoma/leukemia-2,Bcl-2)、半胱氨酸天冬氨酸蛋白水解酶-3(cysteine containing aspartate protein hydrolase-3,Caspase-3)蛋白表达水平的影响;Hoechst 33258染色观察芪白平肺胶囊含药血清对低氧诱导的大鼠PASMCs凋亡的影响。结果20%芪白平肺胶囊含药血清能明显降低低氧诱导的大鼠PASMCs的活力(P<0.05),并显著上调Caspase-3蛋白的表达水平(P<0.05),下调Bcl-2、p-Akt蛋白的表达水平(P<0.05)。加入LY294002后p-Akt蛋白的表达水平下降,且Bcl-2表达水平下降,Caspase-3蛋白表达水平上调(P<0.05)。结论芪白平肺胶囊可能通过抑制PI3K/Akt信号通路促进低氧诱导的大鼠PASMCs凋亡,减缓低氧性肺动脉高压的进程。Objective To investigate the effect of Qibai Pingfei Capsule on hypoxia-induced apoptosis of pulmonary artery smooth muscle cells(PASMCs)in rats via the phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt)pathway at the cellular level.Methods Rat PASMCs were isolated and cultured using the tissue patch method,and drug-containing serum was prepared to act on hypoxia-induced rat PASMCs.PASMCs were divided into normoxia group,hypoxia group,hypoxia+serum containing Qibai Pingfei Capsule group,and hypoxia+serum containing Qibai Pingfei Capsule+LY294002 group.Cell Counting Kit-8 was used to observe the effect of serum containing Qibai Pingfei Capsule on the viability of hypoxia-induced PASMCs;Western blotting was used to investigate the effect of serum containing Qibai Pingfei Capsule alone or in combination with LY294002 on the protein expression levels of Akt,p-Akt,Bcl-2,and caspase-3;Hoechst33258 assay was used to examine the effect of serum containing Qibai Pingfei Capsule on hypoxia-induced apoptosis of rat PASMCs.Results The 20%serum containing Qibai Pingfei Capsule significantly reduced the viability of hypoxia-induced PASMCs(P<0.05),significantly upregulated the protein expression level of caspase-3,and significantly downregulated the protein expression levels of Bcl-2 and p-Akt(P<0.05).After the addition of LY294002,there were significant reductions in the protein expression levels of p-Akt and the anti-apoptotic protein Bcl-2,and there was a significant increase in the protein expression level of caspase-3(P<0.05).Conclusion Qibai Pingfei Capsule may promote the hypoxia-induced apoptosis in PASMCs by inhibiting the PI3K/Akt signaling pathway,thereby slowing down the progression of hypoxic pulmonary hypertension.

关 键 词：芪白平肺胶囊 肺动脉高压 慢性阻塞性肺疾病 血清药理学 磷脂酰肌醇3激酶 蛋白激酶B 凋亡 

分 类 号：R544.16[医药卫生—心血管疾病]