三七总皂苷通过ADAM10/Notch3信号通路干预大鼠PASMCs增殖  

The role of ADAM10/Notch3 signaling pathway in the proliferation of rat PASMCs and intervention of total saponins of Panax notoginseng

作　　者：黄曼 白相书 田云娜 徐俊鹏 王肖婷 张赛 袁琳波 王万铁 HUANG Man;BAI Xiangshu;TIAN Yunna;XU Junpeng;WANG Xiaoting;ZHANG Sai;YUAN Linbo;WANG Wantie(Department of Pathology and Pathophysiology,School of Basic Medicine,Wenzhou Medical University,Wenzhou 325035,Zhejiang,China;Public Health Department of Pingyang County People's Hospital,Pingyang 325400,Zhejiang,China;Department of Functions,School of Basic Medicine,Wenzhou Medical University,Wenzhou 325035,Zhejiang,China)

机构地区：[1]温州医科大学基础医学院病理学与病理生理学系,浙江温州325035 [2]平阳县人民医院公共卫生科,浙江平阳325400 [3]温州医科大学基础医学院机能学系,浙江温州325035

出　　处：《中国临床药理学与治疗学》2025年第4期487-492,共6页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：浙江省介入肺脏病重点实验室建设项目(2019E10014);温州市基础性科研项目(2024Y3124)。

摘　　要：目的:探讨在野百合碱(MCT)作用下三七总皂苷(PNS)对大鼠肺动脉平滑肌细胞(PASMCs)增殖的干预作用及其机制。方法:体外培养的PASMCs随机分为正常对照(Control)组、野百合碱(MCT)组、三七总皂苷(PNS)组、敲低(M+Si ADAM10)组、敲低后处理(M+P+Si ADAM10)组、过表达(M+OE ADAM10)组和过表达后处理(M+P+OE ADAM10)组。造模结束后,采用CCK-8法测各组细胞活力;蛋白质免疫印迹(Western blot)法分别检测细胞增殖细胞核抗原(PCNA)、解整合素金属蛋白酶10(ADAM10)、细胞神经源性基因座notch同源蛋白-3(Notch3)蛋白的表达情况。结果:在MCT作用下,PASMCs活力显著增强(P<0.05或P<0.01);0~400 mg/L的PNS对正常细胞活力无毒性,100 mg/L的PNS可显著抑制MCT诱导的细胞活力(P<0.01)。在敲低ADAM10后PASMCs活力显著减弱(P<0.01),PCNA蛋白表达明显下降(P<0.05),尤以M+P+Si ADAM10组为著;ADAM10、Notch3蛋白表达均显著下降(P<0.05或P<0.01),尤以M+P+Si ADAM10组为著。过表达ADAM10后PASMCs活力显著增强(P<0.01),PCNA蛋白表达明显增高(P<0.01),M+P+OE ADAM10组PCNA值稍偏高(P>0.05),ADAM10、Notch3蛋白表达均显著升高(P<0.05);而过表达ADAM10的同时使用PNS的PASMCs与敲低ADAM10的PASMCs相比,PCNA蛋白表达显著下降(P<0.01),ADAM10、Notch3蛋白表达不同程度降低(P>0.05)。结论:PNS可能通过抑制ADAM10/Notch3信号通路,减弱大鼠MCT诱导的PASMCs增殖。AIM:To investigate the effect and mechanism of panax notoginseng saponins(PNS)inhibiting the proliferation of pulmonary artery smooth muscle cells(PASMCs)in rats under the effect of monocrotaline(MCT).METHODS:PASMCs cultured in vitro were randomly divided into the normal control(Control)group,the monocrotaline(MCT)group,the panax notoginseng saponins(PNS)group,the knockdown(M+Si ADAM10)group,the knockdown postconditioning(M+P+Si ADAM10)group,the overexpression(M+OE ADAM10)group,and the overexpression postconditioning(M+P+OE ADAM10)group.After the model was constructed,the cell viability of each group was measured using the CCK-8 assay,along with Western blot utilized to detect the expression of proliferating cell nuclear antigen(PCNA),disintegrin metalloproteinase 10(ADAM10),and notch homology protein-3(Notch3)at the cellular neurogenic locus,respectively.RESULTS:Under the effect of MCT,the viability of PASMCs was significantly enhanced(P<0.05 or P<0.01);0-400 mg/L PNS was not toxic to the viability of normal cells,and 100 mg/L PNS could significantly inhibit the MCT-induced viability(P<0.01).After the knockdown of ADAM10,the viability of PASMCs significantly declined(P<0.01),and the expression of PCNA protein was significantly decreased(P<0.05),evidently in the M+P+Si ADAM10 group.Meanwhile,the expression of ADAM10 and Notch3 protein was significantly decreased(P<0.05 or P<0.01),evidently in the M+P+Si ADAM10 group.After overexpression of ADAM10,the viability of PASMCs was significantly enhanced(P<0.01),the expression of PCNA protein was significantly increased(P<0.01),the PCNA value was slightly higher(P>0.05),and the expression of ADAM10 and Notch3 protein was significantly elevated(P<0.05)in the M+P+OE ADAM10 group.Additionally,PASMCs overexpressing ADAM10 with concomitant PNS exhibited a significant decrease in the expression of PCNA protein compared with PASMCs knocking down ADAM10(P<0.01),and the expression of ADAM10 and Notch3 protein declined to varying degrees(P>0.05).CONCLUSION:Panax notoginseng saponi

关 键 词：肺动脉平滑肌细胞 三七总皂苷 野百合碱 ADAM10/Notch3通路 大鼠 

分 类 号：R965.2[医药卫生—药理学]