摘要
目的探究滋肾活血方联合低分子肝素(LMWH)对人血管内皮细胞的血管内皮生长因子(VEGF)、血管内皮生长因子受体2(VEGFR-2)、细胞外调节蛋白激酶(ERK)表达及血管生成的影响,探讨其治疗复发性流产的可能作用机制。方法2020年1—12月期间购置人血管内皮细胞试验样本。制备滋肾活血方含药血清和正常血清,分为3组:对照组、LMWH组、滋肾活血方+LMWH组。其对应的处理方式分别为:正常血清、正常血清+LMWH、含药血清+LMWH分别孵育人血管内皮细胞48 h后,应用蛋白质印迹法(western blot)检测各组细胞VEGF、VEGFR-2、ERK蛋白表达量变化,实时荧光定量PCR检测VEGF、VEGFR-2、ERK基因的mRNA转录水平,应用成管实验检测各组细胞的成管能力变化。结果与对照组比较,LMWH组VEGF、VEGFR-2、ERK蛋白的表达水平显著升高(P<0.05),血管生成数和长度均显著升高(P<0.05),VEGF mRNA表达水平显著上调(P<0.05),VEGFR-2、ERK mRNA表达水平比较,差异无统计学意义(P>0.05);与对照组比较,滋肾活血方+LMWH组VEGF、VEGFR-2、ERK mRNA及其蛋白的表达水平显著上调(P<0.05),血管生成数和长度均显著升高(P<0.05);滋肾活血方+LMWH组较LMWH组VEGF、VEGFR-2、ERK蛋白其表达水平升高(P<0.05),血管生成数和长度均增加(P<0.05),VEGF、VEGFR-2、ERK mRNA表达水平比较,差异无统计学意义(P>0.05)。结论滋肾活血方联合LMWH可促进血管生成改善复发性流产患者妊娠结局,其机制可能与调节VEGF、VEGFR-2、ERK表达,促进血管内皮细胞生长有关。
Objective To explore the effects of Zishen Huoxue Recipe combined with low molecular weight heparin(LMWH)on the expression of vascular endothelial growth factor(VEGF),vascular endothelial growth factor receptor 2(VEGFR-2)and extracellular regulatory protein kinase(ERK)in human vascular endothelial cells and the effects on angiogenesis,so as to investigate its possible mechanism of action in the treatment of recurrent miscarriage.Methods The period from January to December 2020 was selected as the research time period,during which the test samples of human vascular endothelial cells were purchased.The medicated serum of Zishen Huoxue Recipe and the normal serum were prepared,and the test samples were divided into three groups:control group,LMWH group,and Zishen Huoxue Recipe+LMWH group.The corresponding treatment methods were:normal serum,normal serum+LMWH,and medicated serum+LMWH.After the human vascular endothelial cells were incubated for 48 hours,Western blot was used to detect the changes in the expression of VEGF,VEGFR-2,and ERK proteins in each group.Real-time fluorescent quantitative PCR was used to detect the mRNA transcription levels of VEGF,VEGFR-2,and ERK genes.The tube formation experiment was used to detect the changes in the tube formation ability of each group of cells.Results Compared with the control group,the expression levels of VEGF,VEGFR-2 and ERK protein in the LMWH group were significantly increased(P<0.05).The number and length of angiogenesis were significantly up-regulated(P<0.05),and the expression level of VEGF mRNA was significantly up-regulated(P<0.05).There was no statistically significant difference in the expression levels of VEGFR-2 and ERK mRNA(P>0.05);compared with the control group,the expression levels of VEGF,VEGFR-2,ERK mRNA and their protein in the Zishen Huoxue Recipe+LMWH group were significantly up-regulated(P<0.05).The number and length of angiogenesis increased significantly(P<0.05);compared with the LMWH group,the expression levels of VEGF,VEGFR-2 and ERK protein in the Zishen Huoxue Recipe+LMWH group were higher(P<0.05),and both the number and length of angiogenesis increased(P<0.05).The expression levels of VEGF,VEGFR-2 and ERK mRNA were not significantly different(P>0.05).Conclusion Zishen Huoxue Recipe combined with LMWH can promote angiogenesis and improve the pregnancy outcome of patients with recurrent miscarriage.The mechanism may be related to regulating the expression of VEGF,VEGFR-2 and ERK and promoting the growth of vascular endothelial cells.
作者
宋宇可
叶利群
黄钟威
张晓芳
吴萍
杨脂
沈颖
SONG Yuke;YE Liqun;HUANG Zhongwei;ZHANG Xiaofang;WU Ping;YANG Zhi;SHEN Ying(The Third School of Clinical Medicine,Zhejiang Chinese Medical University,Hangzhou 310053,China;Department of Gynecology,Ningbo Hospital of Chinese Medicine,Ningbo 315000,China)
出处
《中国现代医生》
2022年第3期59-62,66,F0003,共6页
China Modern Doctor
基金
浙江省“十三五”中医药重点专科建设项目(浙卫办中医〔2019〕1号)
浙江省宁波市科技计划项目(2019 A610358)。
作者简介
通讯作者:宋宇可。
