四五护脉汤对同型半胱氨酸损伤后人脐静脉内皮细胞的保护机制探讨

徐孟娇; 袁文雯; 王雨晴; 徐杰; 金潇; 原方; 韩冉; 杨宝钟; 张凡帆

doi:10.16025/j.1674-1307.2022.11.009

您当前的位置：

首页 >

文章列表页 >

四五护脉汤对同型半胱氨酸损伤后人脐静脉内皮细胞的保护机制探讨

实验研究 | 更新时间：2023-02-15

- 四五护脉汤对同型半胱氨酸损伤后人脐静脉内皮细胞的保护机制探讨
- Exploration of the protective mechanism of Siwu Humai Decoction on homocysteine-induced injury in HUVEC
- 北京中医药 2022年41卷第11期页码：1245-1251
- 作者机构：
  
  1.北京中医药大学东方医院外四科，北京 100078
  2.北京中医药大学东方医院检验科，北京 100078
- 作者简介：
  
  徐孟娇，女，24岁，硕士研究生。研究方向：周围血管疾病的中医诊疗。
  张凡帆，E-mail：fanfandoc@163.com
- 基金信息：
  
  北京中医药大学青年教师项目(2019-JYB-JS-150)
- DOI：10.16025/j.1674-1307.2022.11.009
  中图分类号：
扫描看全文
徐孟娇，袁文雯，王雨晴，等.四五护脉汤对同型半胱氨酸损伤后人脐静脉内皮细胞的保护机制探讨［J］.北京中医药，2022，41（11）：1245-1251.

XU Meng-jiao,YUAN Wen-wen,WANG Yu-qing,et al.Exploration of the protective mechanism of Siwu Humai Decoction on homocysteine-induced injury in HUVEC[J]. Beijing Journal of Traditional Chinese Medicine,2022,41(11):1245-1251.
徐孟娇，袁文雯，王雨晴，等.四五护脉汤对同型半胱氨酸损伤后人脐静脉内皮细胞的保护机制探讨［J］.北京中医药，2022，41（11）：1245-1251. DOI： 10.16025/j.1674-1307.2022.11.009.

XU Meng-jiao,YUAN Wen-wen,WANG Yu-qing,et al.Exploration of the protective mechanism of Siwu Humai Decoction on homocysteine-induced injury in HUVEC[J]. Beijing Journal of Traditional Chinese Medicine,2022,41(11):1245-1251. DOI： 10.16025/j.1674-1307.2022.11.009.

摘要

目的,2,观察四五护脉汤对同型半胱氨酸（homocysteinemia，HCY）诱导人脐静脉内皮细胞（human umbilical vein endothelial cell，HUVEC）损伤的影响及机制。,方法,2,将HUVEC常规培养传代后，MTT法检测细胞活力并确定药物的最佳实验浓度。将细胞分组，A组为正常HUVEC，B~F组加入125 μmol/L的HCY 100 μL制备细胞损伤模型，培养48 h后，C组加入31.25 μmol/L NaHS 100 μL，D、E、F组分别加入1.6、2.2、2.8 g/L四五护脉汤100 μL，继续培养48 h。MTT法检测细胞活力，流式细胞术检测细胞凋亡，Transwell法检测细胞迁移，硝酸还原酶法检测细胞培养液NO水平，Western blot法检测细胞磷脂酰肌醇3-激酶-蛋白激酶B（PI3K）、磷酸化AKT蛋白（p-AKT）抗体、小窝蛋白-1（Caveolin-1，CAV-1）、内皮型一氧化氮合酶（endothelial nitric oxide synthase，eNOS）蛋白表达，RT-PCR法检测细胞内eNOS mRNA表达，BCA测定细胞黏附率。,结果,2,HCY、NaHS最佳实验浓度分别为125、31.25 μmol/L，四五护脉汤干预浓度为1.6、2.2、2.8 g/L。HCY干预后，HUVEC活力下降，凋亡率上升，迁移率下降，NO水平降低，eNOS、PI3K、p-AKT蛋白相对表达量降低，CAV-1蛋白相对表达量升高，eNOS mRNA相对表达量降低，单核细胞黏附率升高（,P,<,0.05）；而四五护脉汤可逆转上述变化（,P,<,0.05），且呈浓度依赖性。,结论,2,四五护脉汤能够通过调节PI3K-AKT-eNOS信号通路，提高NO表达，减少内皮细胞凋亡，提高内皮细胞活力、迁移，抑制单核细胞黏附，从而减少内皮细胞损伤，达到保护内皮细胞的目的。

Abstract

Objective,2,To observe the effects and mechanism of Siwu Humai Decoction on the injury of human umbilical vein endothelial cells （HUVEC） induced by homocysteine （HCY）.,Methods,2,After conventional culture and passage of HUVEC，MTT assay was used to detect the cell viability and determine the best experimental concentration of the drug. The cells were divided into groups： group A was normal HUVEC， group B to F was added with 125 μmol/L HCY 100 μL to make cell injury model， after 48 hours of culture， group C was added with 31.25 μmol/L NaHS 100 μL， and groups D， E and F were added with 1.6， 2.2 and 2.8 g/L Siwu Humai Decoction 100 μL respectively， and continued to culture.MTT method was used to detect cell viability of each group， flow cytometry to detect apoptosis， transwell method to detect cell migration rate in each group；nitrate reductase method to detect NO level in cell culture medium. Western blot to detect PI3K， p-AKT， CAV-1， eNOS protein relative expression in each group， and BCA measurement of endothelial cell-monocyte adhesion rate.,Results,2,The optimal concentrations of HCY and NaHS were 125 μmol/L and 31.25 μmol/L， respectively. The intervention concentration of Siwu Humai Decoction was selected as 1.6， 2.2， and 2.8 g/L. After HCY intervention， the cell viability values were decreased， apoptosis rate increased， migration rate decreased， NO level decreased，eNOS，PI3K，p-AKT protein relative expression decreased， CAV-1 protein relative expression increased， eNOS mRNA relative ct value decreased and adhesion rate with monocytes increased， while Siwu Humai Decoction could reverse the above expressions（,P,<,0.05），and is concentration-dependent.,Conclusion,2,Siwu Humai Decoction can protect HUVEC after HCY injury by regulating the PI3K-AKT-eNOS signaling pathway， increasing NO expression， reducing endothelial cell apoptosis， and improving endothelial cell viability and migration， inhibiting monocyte adhesion， thus reducing endothelial cell damage and achieving the purpose of protecting endothelial cells.

关键词

深静脉血栓四五护脉汤同型半胱氨酸内皮细胞PI3K-AKT-eNOS信号通路

Keywords

Deep vein thrombosisSiwu Humai Decoctionhomocysteineendothelial cell functionPI3K-AKT-eNOS signaling pathway

references

DI NISIO M, VAN ES N, BÜLLER HR. Deep vein thrombosis and pulmonary embolism[J]. Lancet, 2016,388(10063):3060-3073.

KÖKTÜRK N, KANBAY A, AYDOĞDU M, et al. Hyperhomocysteinemia prevalence among patients with venous thromboembolism[J]. Clin Appl Thromb Hemost, 2011,17(5):487-493.

WOLBERG AS, ROSENDAAL FR, WEITZ JI, et al. Venous thrombosis[J]. Nat Rev Dis Primers, 2015,1:15006.

张伟, 贺冰, 李亮,等. 补阳还五汤促进内皮祖细胞修复损伤血管内皮[J]. 中国病理生理杂志,2017,33(11):1969-1974.

刘湘, 张玥, 公茂旺,等. 基于网络药理学的四妙勇安汤治疗深静脉血栓形成机制研究[J]. 海南医学院学报,2021,27(11):855-860,867.

MOLL S, VARGA EA. Homocysteine and MTHFR mutations[J]. Circulation, 2015,132(1):e6-9.

林晶, 李友山, 张凡帆,等. 庞鹤教授运用虫类药治疗下肢深静脉血栓经验总结[J]. 现代中医临床,2018,25(4):36-39.

LAI WK, KAN MY. Homocysteine-induced endothelial dysfunction[J]. Ann Nutr Metab, 2015,67(1):1-12.

FU Y, WANG X, KONG W. Hyperhomocysteinaemia and vascular injury: advances in mechanisms and drug targets[J]. Br J Pharmacol, 2018,175(8):1173-1189.

YOSHIOKA K, YOSHIDA K, CUI H, et al. Endothelial PI3K-C2α, a class II PI3K, has an essential role in angiogenesis and vascular barrier function[J]. Nat Med, 2012,18(10):1560-1569.

FÖRSTERMANN U, XIA N, LI H. Roles of vascular oxidative stress and nitric oxide in the pathogenesis of atherosclerosis[J]. Circ Res, 2017,120(4):713-735.

ZHANG Y, JANSSENS SP, WINGLER K, et al. Modulating endothelial nitric oxide synthase: a new cardiovascular therapeutic strategy[J]. Am J Physiol Heart Circ Physiol, 2011,301(3):H634-646.

LIN F, YANG Y, WEI S, et al. Hydrogen sulfide protects against high glucose-induced human umbilical vein endothelial cell injury through activating PI3K/Akt/eNOS pathway[J]. Drug Des Devel Ther, 2020,14:621-633.

VON BRÜHL ML, STARK K, STEINHART A, et al. Monocytes, neutrophils, and platelets cooperate to initiate and propagate venous thrombosis in mice in vivo[J]. J Exp Med, 2012,209(4):819-835.

MAO XH, TAO SU, ZHANG XH. Apelin-13 promotes monocyte adhesion to human umbilical vein endothelial cell-mediated by the phosphatidylinositol 3-kinase signaling pathway[J]. Prog Biochem Biophys, 2011,38:1162-1170.

尤心怡, 王文凤, 徐霞,等. 补阳还五汤治疗动脉粥样硬化的物质基础及作用机制研究进展[J]. 中药新药与临床药理,2021,32(10):1575-1581.

谢盈彧,刘璐,李渊芳,等.基于网络药理学的四妙勇安汤在血管新生中的作用机制研究[J].中草药,2018,49(18):4319-4330.

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

暂无数据