摘要
目的通过观察马兜铃酸-I(AA-I)对人脐静脉内皮细胞(HUVEC)增殖、凋亡、分泌、转分化的影响,探讨其与共刺激分子CD40/CD40L、转化生长因子-β1(TGF-β1)/TGF-βⅡ型受体(TGF-βⅡR)的关系。方法应用不同刺激浓度的AA-I作用于HUVEC,分别采用四甲基偶氮唑盐(MTT)法和流式细胞仪(FCM)检测AA-I对HUVEC增殖和凋亡的影响;FCM检测血管内皮细胞的标志分子血管内皮生长因子受体(Flk1),共刺激分子CD40L、促纤维化细胞因子TGF-βⅡR表达的变化;Western blot法测定肌成纤维细胞的标志蛋白α-平滑肌肌动蛋白(α-SMA)、CD40、TGF-β1表达的变化;酶联免疫吸附法检测HUVEC分泌TGF-β1的变化。结果不同浓度AA-I作用于HUVEC培养96 h后,高浓度AA-I导致细胞增殖受到抑制,各刺激浓度均可导致凋亡,Flk1表达下调、α-SMA表达上调,共刺激分子CD40/CD40L、TGF-β1/TGF-βⅡR表达增加,明显促进TGF-β1的分泌(P<0.05),上述实验数据改变均呈明显的剂量相关性。结论 AA-I不仅能够抑制内皮细胞增殖,促进其凋亡,并能使其向平滑肌样细胞转化和纤维化。
Objective To investigate the effects of aristolochic acid-I（AA-I） on the human umbilical vein endothelial cell（HUVEC） by studying its proliferation,apoptosis,secretion and transdifferentiation,and explore its connection with costimulatory molecules CD_40/CD_40L and transforming growth factor-β1（TGF-β1）/TGF-βⅡ receptor（TGF-βⅡR）. Methods HUVEC was treated with various doses of AA-I,then HUVEC proliferation and apoptosis were detected by methyl thiazolyl tetrazolium（MTT） and flow cytometry（FCM）,respectively.The expression variance of endothelial cell marker vein endothelial growth factor receptor（Flk1）,costimulatory molecules CD_40L and promotion fibrosis cytokine TGF-βⅡR were detected by FCM.The expression variance of the muscle fibroblast marker protein α-smooth muscle actin（α-SMA）,CD_40 and TGF-β1 were detected by Western blot;TGF-β1 level was measured by enzyme-linked immunosorbent assay. Results After HUVEC were treated with different concentration AA-I for 96 hours,cell proliferations were inhibited by high concentration of AA-I.The apoptosis was induced at each concentration.Down regulated Flk1 and up regulated α-SMA expression were found.The expressions of CD_40/CD_40L,TGF-β1/TGF-βⅡR were enhanced,and TGF-β1 secretion was promoted（P0.05）.All results referred were dose-related apparently. Conclusion AA-I could induce the endothelial cells growth inhibition,apoptosis and make the endothelial cells transferred to smooth muscle like cell and fibrosis.
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2011年第5期316-318,321,共4页
Journal of Applied Clinical Pediatrics
基金
国家自然科学基金(30271234)
