摘要
背景:关于超敏C-反应蛋白水平正常者血清同型半胱氨酸浓度与冠状动脉病变的关系研究显示,在超敏C-反应蛋白水平正常者中,血清同型半胱氨酸浓度是冠状动脉疾病患病率和严重程度的独立预测因子。目的:探讨稳斑汤调控PI3K/Akt信号通路对同型半胱氨酸诱导大鼠心脏微血管内皮细胞凋亡的保护作用。方法:体外原代培养大鼠心脏微血管内皮细胞,分为对照组、模型组(用10 mmol/L同型半胱氨酸诱导细胞损伤)、稳斑汤组(同型半胱氨酸+50 mg/L稳斑汤给药)。CCK-8检测各组细胞存活能力;ELISA检测各组细胞上清中乳酸脱氢酶、丙二醛、全血氧化氢酶、超氧化物歧化酶、谷胱甘肽过氧化物酶、白细胞介素6、细胞间黏附分子1、白细胞介素1β、肿瘤坏死因子α含量;并在稳斑汤治疗基础上加入了PI3K抑制剂LY294002,流式细胞术检测各组细胞凋亡情况;Western blot法检测各组细胞中PI3K、Akt、p-Akt、Bax、Bcl-2和caspase3蛋白水平的表达。实验获得辽宁中医药大学动物实验伦理委员会批准(批准号为21000092018010)。结果与结论:①与对照组相比,模型组心脏微血管内皮细胞存活能力显著降低,乳酸脱氢酶的渗漏量显著增加,引起氧化应激以及炎症因子的释放,最终导致细胞大量凋亡,差异有显著性意义;与模型组相比,稳斑汤组心脏微血管内皮细胞存活能力增加,乳酸脱氢酶的渗漏量降低,细胞中白细胞介素1β、细胞间黏附分子1、白细胞介素6和肿瘤坏死因子α的浓度降低(P<0.05),细胞凋亡数目减少;②PI3K抑制剂能够逆转稳斑汤对同型半胱氨酸诱导的心脏微血管内皮细胞凋亡的抑制作用;③结果说明,稳斑汤可显著提高同型半胱氨酸损伤的心脏微血管内皮细胞存活能力,降低乳酸脱氢酶的渗漏量,抑制氧化应激水平以及炎症因子的释放,最终减少细胞凋亡数,这种作用与抑制PI3K/Akt信号通路有关。
BACKGROUND:Studies have shown that serum homocysteine concentration is an independent predictor of the prevalence and severity of coronary artery disease for patients with normal hypersensitivity C-reactive protein levels.OBJECTIVE:To investigate the protective effect of Wenban Decoction on the apoptosis of rat cardiac microvascular endothelial cells(CMECs)induced by homocysteine by regulating PI3K/Akt signaling pathway.METHODS:Rat CMECs were primarily cultured in vitro,and the cells were randomly divided into control group,model group and Wenban Decoction group(50 mg/L Wenban Decoction).The cells in the latter two groups were injured by 10 mmol/L homocysteine prior to the treatment.Cell counting kit-8 was used to detect the cell viability of each group.ELISA was used to determine serum lactate dehydrogenase,malondialdehyde,whole blood catalase,superoxide dismutase,glutathione peroxidase,interleukin 6,intercellular adhesion molecule 1,interleukin 1β,and tumor necrosis factorα.Flow cytometry was used to detect cell apoptosis after addition of LY294002 based on the treatment with Wenban Decoction.Western blot was used to detect the expression of PI3K,Akt,p-Akt,Bax,Bcl-2 and caspase3 protein in the cells.An ethic approval was given by the Animal Experiment Ethics Committee of Liaoning University of Traditional Chinese Medicine(approval No.21000092018010).RESULTS AND CONCLUSION:Compared with the control group,the survival ability of CMECs in the model group was significantly reduced,the leakage of lactate dehydrogenase was significantly increased,which caused oxidative stress and the release of inflammatory factors,and finally led to a large number of apoptosis.Compared with the model group,Wenban Decoction improved the survival ability of CMECs,reduced the leakage of lactate dehydrogenase,significantly decreased the intracellular levels of interleukin 1β,intercellular adhesion molecule 1,interleukin 6 and tumor necrosis factorα(P<0.05),as well as reduced the number of apoptotic cells.PI3K inhibitor reversed the
作者
张雯雯
金颂峰
赵国梁
宮丽鸿
Zhang Wenwen;Jin Songfeng;Zhao Guoliang;Gong Lihong(Liaoning University of Traditional Chinese Medicine,Shenyang 110032,Liaoning Province,China;Third Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,Liaoning Province,China;First Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,Liaoning Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2021年第5期723-728,共6页
Chinese Journal of Tissue Engineering Research
基金
辽宁省科学技术基金项目(20180530016),项目负责人:宮丽鸿
辽宁特聘教授课题项目(辽委发[2017]3号),项目负责人:宮丽鸿。
关键词
冠心病
心脏
微血管
内皮细胞
细胞凋亡
通路
中药
实验
coronary heart disease
heart
microvessel
endothelial cell
apoptosis
pathway
Chinese herb
experiment
