来源：风湿病与关节炎,2020,9（2）:1-6,17.

化痰通络方含药血清对血管内皮生长因子诱导的脐静脉内皮细胞增殖的影响及其机制研究（基础研究）

邱明山¹，陈进春¹，林双杰¹，钱丽霞²，张少红³，李良成²

　　【摘　要】目的：观察化痰通络方含药血清对血管内皮生长因子（VEGF）诱导的脐静脉内皮细胞（HUVEC）增殖的影响及其机制。方法：将20只新西兰兔随机分为化痰通络方高（HTTL-h）、中（HTTL-m）、低（HTTL-l）剂量组，雷公藤多苷组（TG组），生理盐水组（Sal组），每组4只。分离鉴定健康新生儿HUVEC，siRNA敲降VEGFR2、PLCG1以及MAPK1，并采用RT-PCR和及Western Blot检测VEGFR2、PLCG1和MAPK1 mRNA 及蛋白水平的表达。MTT法检测经VEGF诱导的HUVEC增殖情况。结果：与Sal组比较，VEGF可显著促进HUVEC增殖，而TG或HTTL-l均可显著抑制VEGF诱导的HUVEC增殖；siRNA敲降VEGFR2后，VEGF则不能促进HUVEC增殖，同时TG也失去了对HUVEC增殖的抑制能力；siRNA敲降PLCG1后，VEGF依然可以显著促进HUVEC增殖，此时TG对VEGF刺激的HUVEC增殖失去了抑制效应，但HTTL-m和HTTL-l依然对VEGF刺激的HUVEC增殖具有显著抑制效应；siRNA敲降MAPK1后，VEGF也可显著促进HUVEC增殖，TG对VEGF刺激的HUVEC增殖具有抑制效应，同时HTTL-l也对VEGF刺激的HUVEC增殖具有显著抑制效应。结论：化痰通络方含药血清对VEGF诱导的HUVEC增殖具有抑制作用。TG抑制VEGF诱导的HUVEC增殖依赖于VEGFR2和PLCG1表达，而化痰通络方对VEGF刺激的HUVEC增殖的抑制效应则不依赖VEGF/VEGFR2/PLCG1/MAPK1信号通路。

　　【关键词】　关节炎，类风湿；化痰通络方；血管内皮生长因子；脐静脉内皮细胞；增殖；信号传导通路

On the Effects of Serum Containing Huatan Tongluo Fang（化痰通络方）on the Proliferation of HUVEC Induced by VEGF and Its Mechanism

QIU Ming-shan,CHEN Jin-chun,LIN Shuang-jie,QIAN Li-xia,ZHANG Shao-hong,LI Liang-cheng

　　【ABSTRACT】Objective:To observe the effects of serum containing Huatan Tongluo Fang（HTTL,化痰通络方）on the proliferation of HUVEC induced by VEGF and its mechanism.Methods:Twenty New Zealand rabbits were randomly divided into the High（HTTL-h）,medium（HTTL-m）,low（HTTL-l） dose groups,the Tripterygium wilfordii glycoside group（TG group） and the saline containing serum group（Sal group）,4 rabbits in each group.Healthy neonates HUVEC were isolated and identified,and siRNA knocked down VEGFR2,PLCG1,and MAPK1.RT-PCR as well as Western Blot were used to detect the expressions of VEGFR2,PLCG1,MAPK1 mRNA and protein levels.The proliferation of HUVEC induced by VEGF was detected by MTT.Results:Compared with the Sal group,VEGF could significantly promote HUVEC prolifer-ation,while TG or HTTL-l could significantly inhibit HUVEC proliferation induced by VEGF.After siRNA knocked down VEGFR2,VEGF could not promote HUVEC proliferation,while TG also lost the inhibition of HUVEC proliferation.After siRNA knocked down PLCG1,VEGF could still significantly promote HUVEC proliferation,and at this time TG could not stimulate HUVEC proliferation stimulated by VEGF.However,HTTL-m and HTTL-l still had a significant inhibitory effect on HUVEC proliferation stimulated by VEGF.After siRNA knocked down MAPK1,VEGF could also significantly promote HUVEC proliferation,TG had a inhibitory effect on HUVEC proliferation stimulated by VEGF,and HTTL-l also had a significant inhibitory effect on HUVEC proliferation stimulated by VEGF.Conclusion:The serum containing HTTL can inhibit the proliferation of HUVEC induced by VEGF.In TG group,the inhibition of HUVEC proliferation induced by VEGF depended on the expression of VEGFR2 and PLCG1,while the inhibition of HTTL on HUVEC proliferation stimulated by VEGF did not depend on the VEGF/VEGFR2/ PLCG1/ MAPK1 signal pathway.

　　【Keywords】 arthritis,rheumatoid;Huatan Tongluo Fang（化痰通络方）;VEGF;HUVEC;proliferation;signal transduction pathway