摘要
目的明确激活素A(ACT A)对成人脐静脉内皮细胞转分化的影响及卵泡抑素(FS)的干预作用。方法体外培养成人脐静脉内皮细胞(HUVEC),MTT法检测不同浓度rh?ACT A、rh?FS刺激内皮细胞后的增殖率变化。将细胞分为5组:对照组、TGF?β组(T组)、ACT A组(A组)、TGF?β+FS组(T+F组)、ACT A+FS组(A+F组),采用细胞化学染色和Western Blot蛋白印迹技术观察ACT A和TGF?β对HUVEC细胞α?平滑肌动蛋白(α?SMA)、纤维连接蛋白(FN)和VE?钙黏蛋白(VE?cad)表达的影响以及FS的干预作用。结果与0 ng/mL ACT A组相比,10、30、60、100 ng/mL ACT A组的增殖活性均增加(F=22.768,P<0.001),其中30 ng/mL ACT A组的增殖率最高,单独给予FS对细胞增生无明显影响(F=0.437,P=0.781),但FS能拮抗ACT A对内皮细胞生长的促进作用,呈剂量依赖性(F=15.270,P<0.001)。细胞化学染色和Western Blot蛋白印迹结果显示:ACT A组与对照组相比,α?SMA和FN的表达增加,而VE?Cad的表达减少(均P<0.05),与TGF?β组相比,α?SMA、FN和VE?Cad的表达差异无统计学意义(均P>0.05);FS可阻断ACT A的上述效应(P<0.05),而对TGF?β的作用无明显影响(P>0.05)。ACT A+FS组与ACT A组相比,α?SMA和FN的表达减少,同时VE?Cad的表达增加(均P<0.05)。TGF?β+FS组与TGF?β组比较,α?SMA、FN和VE?Cad的表达差异无统计学意义(均P>0.05)。结论ACT A可诱导内皮细胞转分化及FN的合成,FS可通过阻断ACT A的上述效应,产生内皮细胞保护作用。
Objective To investigate the effect of activin A(ACT A)on the transdifferentiation of human umbilical vein endothelial cells and the intervention of follistatin(FS).Methods Human umbilical vein endothelial cells(HUVEC)were cultured in vitro.The proliferation rate of HUVEC stimulated by rh?ACT A and rh?FS was measured by MTT assay.Cells were divided into 5 groups:control group,TGF?β group(group T),ACT A group(group A),TGF?β+FS group(group T+F)and ACT A+FS group(group A+F).The effects of ACT A and TGF?β on the expression of α?smooth muscle actin(α?SMA),fibronectin(FN)and VE?cadherin(VE?cad)in HUVEC cells were observed by cytochemical staining and Western Blot.Results Compared with 0 ng/mL ACT A group,the proliferation activity of 10 ng/mL ACT A group,30 ng/mL ACT A group,60 ng/mL ACT A group and 100 ng/mL ACT A group were increased(F=22.768,P<0.001).The proliferation rate of 30 ng/mL ACT A group was the highest,and FS alone had no significant effect on cell proliferation(F=0.437,P=0.781).However,FS could antagonize the effect of ACT A on the growth of endothelial cells in a dose?dependent manner(F=15.270,P<0.001).Cytochemical staining and Western Blot showed that the expression ofα?SMAand FN was increased,while the expression of VE?Cad was decreased(P<0.05)after stimulation of ACT A on endothelial cell,compared with the control group.Compared with TGF?β group,there was no statistically significant difference in the expression of α?SMA,FN and VE?Cad(P>0.05)in ACT A group.FS could block the effect of ACT C,but had no effect on TGF?β(P>0.05).Compared with ACT A group,the expression of α?SMA and FN was decreased and the expression of VE?Cad was increased in ACT A+FS group(P<0.05).There was no significant difference in the expression ofα?SMA,FN and VE?Cad between TGF?β+FS group and TGF?β group(P>0.05).Conclusion ACT A can induce endothelial cell transdifferentiation and FN synthesis,and FS can block the above effects of ACT A and exerts an endothelial protection.
作者
张湾
刘高虹
于为民
王艳红
张楠
吴之敬
任小军
ZHANG Wan;LIU Gaohong;YU Weimin;WANG Yanhong;ZHANG Nan;WU Zhijing;REN Xiaojun(Department of Nephrology,the Affiliated Hospital of Shanxi Medical University,Taiyuan 030032,China)
出处
《实用医学杂志》
CAS
北大核心
2019年第21期3276-3280,共5页
The Journal of Practical Medicine
基金
国家自然科学基金项目(编号：81500518)
山西省卫计委科研基金项目(编号：2015004,2011005)
关键词
激活素A
卵泡抑素
内皮-间充质转分化
转化生长因子Β
activin A
follicular statin
endothelin mesenchymal transition
transforming growth factorβ
