越婢汤含药血清对阿霉素损伤的人脐静脉血管内皮细胞的干预作用  

Interventional effects of drug‑containing serum of Yuebitang on adriamycin‑injured human umbilical vein vascular endothelial cell

作　　者：李婷婷[1,2,3,4] 程苏 王艳茹 邵命海 LI Tingting;CHENG Su;WANG Yanru;SHAO Minghai(Department of Nephrology,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;TCM Institute of Kidney Disease,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Key Laboratory of Liver and Kidney Diseases,Ministry of Education,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Shanghai Key Laboratory of Traditional Chinese Clinical Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属曙光医院肾病科,上海201203 [2]上海中医药大学中医肾病研究所,上海201203 [3]上海中医药大学肝肾疾病病证教育部重点实验室,上海201203 [4]上海市中医临床重点实验室,上海201203

出　　处：《上海中医药杂志》2024年第3期25-31,共7页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81973807)。

摘　　要：目的基于小窝蛋白-1(Cav-1)/内皮型一氧化氮合酶(eNOS)通路,探讨越婢汤含药血清对阿霉素(ADR)损伤的人脐静脉血管内皮细胞(HUVEC)模型的作用机制。方法制备越婢汤含药血清及空白血清,体外培养HUVEC,采用ADR诱导建立HUVEC损伤模型,将细胞分为对照组、模型组、越婢汤组(2.5%、5%、10%、20%含药血清),给予相应药物处理24 h。细胞增殖-毒性检测(CCK-8)法检测细胞存活率,划痕实验检测细胞迁移能力,Western blot法检测Cav-1/eNOS信号通路相关蛋白表达,免疫荧光法检测细胞Cav-1表达水平,酶联免疫吸附测定(ELISA)法检测细胞上清液Cav-1的含量。结果与对照组比较,模型组细胞增殖及迁移能力降低(P<0.05),细胞内Cav-1、磷酸化内皮型一氧化氮合酶(p-eNOS)、血管内皮细胞钙黏蛋白(VE-cadherin)表达下降(P<0.05),细胞上清液Cav-1含量上升(P<0.05)。与对照组比较,各浓度越婢汤含药血清对HUVEC细胞活力无明显影响(P>0.05)。与模型组比较,越婢汤各组能促进细胞迁移(P<0.05);2.5%浓度的含药血清对细胞Cav-1等蛋白表达无影响(P>0.05);5%、10%、20%浓度含药血清能增加细胞内Cav-1、p-eNOS、VE-cadherin蛋白的表达(P<0.05),下调细胞上清液Cav-1的含量(P<0.05)。结论越婢汤含药血清可以降低ADR对HUVEC细胞的损伤,促进HUVEC细胞的增殖、迁移,其保护作用可能与调控Cav-1/eNOS信号通路有关。Objective To investigate the mechanism of action of serum containing Yuebitang on adriamycin(ADR)-injured human umbilical vein vascular endothelial cell(HUVEC)model based on the caveolin-1(Cav-1)/endothelial-type nitric oxide synthase(eNOS)pathway.Methods The HUVEC were cultured in vitro with the serum containing Yuebitang and blank serum.ADR induction was used to establish the HUVEC injury model,and the cells were divided into control group,model group and the serum containing Yuebitang groups(2.5%,5%,10%and 20%drug-containing serum)and treated with corresponding drugs for 24 h.The cell viability was detected by cell proliferation-toxicity assay(CCK-8),the cell migration ability by the scratch assay,the expression of the Cav-1/eNOS signaling pathway-related proteins by Western blot,the expression level of Cav-1 in the cells by the immunofluorescence assay,and the content of Cav-1 in cell supernatants by ELISA assay.Results Compared with the control group,the proliferation and migration ability of cells in the model group decreased(P<0.05),and the expression of intracellular Cav-1,p-eNOS and VEcadherin proteins decreased(P<0.05),while the content of Cav-1 in the cell supernatant increased(P<0.05).Compared with the control group,there was no significant effect on HUVEC cell viability at all concentrations of Yuebitang containing serum(P>0.05).Compared with the model group,each group of Yuebitang could promote cell migration(P<0.05);2.5%concentration of drug-containing serum had no statistically significant effect on cellular expression of Cav-1 and other proteins(P>0.05);5%,10%,and 20%concentration of drug-containing serum increased the expression of intracellular Cav-1,p-eNOS and VE-cadherin proteins(P<0.05)and down-regulated the content of Cav-1 in cell supernatant(P<0.05).Conclusion The serum containing Yuebitang can reduce the damage of ADR on HUVEC cells and promote the proliferation and migration of HUVEC cells,and its protective effect may be related to the regulation of Cav-1/eNOS signaling pathway.

关 键 词：肾病综合征 越婢汤 经典名方 血管内皮细胞 水肿 作用机制 中药研究 

分 类 号：R285[医药卫生—中药学]