麝香通心滴丸改善AngⅡ介导的血管平滑肌细胞功能紊乱作用机制  

Mechanism of Shexiang Tongxin Dripping Pills in Ameliorating AngⅡ-induced Vascular Smooth Muscle Cell Dysfunction

作　　者：胡玥瑶 王伟 黄明月[1,2] 郭抒娟 姚骏凯 HU Yueyao;WANG Wei;HUANG Mingyue;GUO Shujuan;YAO Junkai(School of Traditional Chinese Medicine(TCM),Beijing University of Chinese Medicine,Beijing 100029,China;Key Laboratory of Basic Research on Syndromes and Prescriptions,Ministry of Education,Beijing University of Chinese Medicine,Beijing 100029,China;College of TCM,Guangzhou University of Chinese Medicine,Guangzhou 510006,China)

机构地区：[1]北京中医药大学中医学院,北京100029 [2]北京中医药大学证候与方剂基础研究教育部重点实验室,北京100029 [3]广州中医药大学中药学院,广州510006

出　　处：《中国实验方剂学杂志》2025年第8期97-106,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81930113,82305216);中国博士后科学基金项目(2023M730345)。

摘　　要：目的:研究麝香通心滴丸含药血清(STDP)缓解血管紧张素Ⅱ(AngⅡ)诱导的细胞表型转化、增殖迁移改善血管平滑肌细胞功能紊乱的作用机制。方法:采用AngⅡ诱导血管平滑肌细胞建立增殖迁移模型,分为空白组(Control)、模型组(Model)、STDP(5%、10%、20%)剂量组干预24 h。通过免疫荧光检测血管α-平滑肌肌动蛋白(α-SMA)、基质金属蛋白酶-2(MMP-2)的表达量。细胞增殖与活性检测(CCK-8)试剂盒、5-乙炔基-2'-脱氧尿嘧啶核苷(EdU)染色检测血管平滑肌细胞增殖情况,划痕实验检测血管平滑肌细胞迁移情况。蛋白免疫印迹法(Western blot)检测血管紧张素转化酶2(ACE2)、血管紧张素Ⅱ受体2(AT2)、血管紧张素Ⅱ受体1(AT1)等通路蛋白,以及Ⅰ型胶原蛋白(ColⅠ)、骨桥蛋白(OPN)等增殖迁移蛋白的表达。结果:与模型组比较,免疫荧光结果表明STDP可明显增加α-SMA表达量,明显抑制MMP-2表达量(P<0.05);CCK-8及EdU染色结果显示STDP明显抑制血管平滑肌细胞增殖(P<0.05);划痕实验检测结果表明STDP明显抑制血管平滑肌细胞迁移(P<0.05);Western blot结果显示STDP可明显提高ACE2、AT2、MAS1的表达量,明显抑制AT1的表达量(P<0.05),明显抑制PCNA、ColⅠ、MMP-9、Rock1、Rock2、SRF蛋白的表达水平(P<0.05)。与STDP组比较,ACE2抑制剂逆转了STDP的调控作用。结论:STDP抑制血管平滑肌细胞表型转化、增殖迁移,并调控细胞增殖迁移相关蛋白的表达,改善血管平滑肌细胞功能紊乱,其机制与提高ACE2-AT2/MAS通路蛋白表达,抑制AT1/Rock信号通路蛋白表达有关。Objective:To study the mechanism of Shexiang Tongxin Dripping pills-containing serum(STDP)in ameliorating angiotensinⅡ(AngⅡ)-induced cell phenotype transformation,proliferation,migration,and dysfunction of vascular smooth muscle cells.Methods:An AngⅡ-induced proliferation and migration model of vascular smooth muscle cells was established.The cells were treated with STDP at 5%,10%,and 20%for 24 h.The immunofluorescence assay was employed to detect the expression ofαsmooth muscle actin(α-SMA)and matrix metalloproteinase-2(MMP-2).The cell-counting kit-8(CCK 8)assay and 5-ethynyl-2'-deoxyuridine(EdU)staining were employed to detect the proliferation of vascular smooth muscle cells,and the scratch assay was employed to detect the migration of the cells.Western blot was employed to determine the expression levels of pathway proteins such as angiotensin-converting enzyme 2(ACE2),angiotensinⅡtype 2(AT2),angiotensinⅡtype 1(AT1),as well as proliferation and migration proteins such as typeⅠcollagen(ColⅠ)and osteopontin(OPN).Results:Compared with the model group,STDP increased the expression ofα-SMA,reduced the expression of MMP-2,and inhibited the proliferation and migration of vascular smooth muscle cells(P<0.05).Furthermore,STDP up-regulated the expression levels of ACE2,AT2,and MAS1,while down-regulating the expression level of AT1,PCNA,ColⅠ,MMP-9,Rock1,Rock2,and SRF(P<0.05).Compared with the STDP group,the ACE2 inhibitor reversed the regulatory effects of STDP.Conclusion:STDP inhibits the phenotype transformation,proliferation,and migration of vascular smooth muscle cells and regulates the expression of cell proliferation and migration-related proteins to ameliorate the dysfunction of vascular smooth muscle cells.It exerts the effects by up-regulating the expression of proteins in the ACE2-AT2/MAS pathway and down-regulating the expression of proteins in the AT1-Rock signaling pathway.

关 键 词：麝香通心滴丸 血管平滑肌细胞功能紊乱 细胞增殖迁移 血管紧张素Ⅱ(AngⅡ)信号通路 

分 类 号：R242[医药卫生—中医临床基础] R289[医药卫生—中医学] R259