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中国临床药理学与治疗学 ›› 2025, Vol. 30 ›› Issue (4): 487-492.doi: 10.12092/j.issn.1009-2501.2025.04.006

• 基础研究 • 上一篇    下一篇

三七总皂苷通过ADAM10/Notch3信号通路干预大鼠PASMCs增殖

黄曼1,白相书2,田云娜1,徐俊鹏1,王肖婷1,张赛1,袁琳波3,王万铁1   

  1. 1温州医科大学基础医学院病理学与病理生理学系,温州  325035,浙江;2平阳县人民医院公共卫生科,平阳  325400,浙江;3温州医科大学基础医学院机能学系,温州  325035,浙江
  • 收稿日期:2024-04-01 修回日期:2024-05-30 出版日期:2025-04-26 发布日期:2025-04-09
  • 通讯作者: 王万铁,男,教授,研究方向:肺动脉高压及缺血再灌注损伤。 E-mail: wwt@wmu.edu.cn 袁琳波,女,副教授,研究方向:肺动脉高压。 E-mail: 81132302@qq.com
  • 作者简介:黄曼,女,硕士研究生,研究方向:肺动脉高压。 E-mail: 1440480267@qq.com 白相书,女,副主任护师,研究方向:肺动脉高压。 E-mail: 2577001560@qq.com
  • 基金资助:
    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)

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

HUANG Man1, BAI Xiangshu2, TIAN Yunna1, XU Junpeng1, WANG Xiaoting1, ZHANG Sai1, YUAN Linbo3, WANG Wantie1   

  1. 1Department of Pathology and Pathophysiology, School of Basic Medicine, Wenzhou Medical University, Wenzhou 325035, Zhejiang, China; 2Public Health Department of Pingyang County People's Hospital, Pingyang 325400, Zhejiang, China; 3Department of Functions, School of Basic Medicine, Wenzhou Medical University, Wenzhou 325035, Zhejiang, China
  • Received:2024-04-01 Revised:2024-05-30 Online:2025-04-26 Published:2025-04-09

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。

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

Abstract:

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 saponins can mitigate MCT-induced PASMCs proliferation in rats by inhibiting the ADAM10/Notch3 signaling pathway.

Key words: pulmonary artery smooth muscle cells, panax notoginseng saponins, monocrotaline, ADAM10/Notch3 pathway, rats

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