PCA通过NF-κB和AP-1信号通路影响HUVECs表达TF  

PCA affects expression of TF in HUVECs through NF-κB and AP-1 signaling pathways

作　　者：靳文 张根葆[1,2] 高恬媛 桑金凤 JIN Wen;ZHANG Gen-bao;GAO Tian-yuan;SANG Jm-feng(Department of Pathophysiology, 2Snake Venom Research hlsttitute, Wannan Medical College, Wuhu 241002, China)

机构地区：[1]皖南医学院病理生理学教研室,安徽芜湖241002 [2]皖南医学院蛇毒研究所,安徽芜湖241002

出　　处：《中国病理生理杂志》2018年第5期904-908,共5页Chinese Journal of Pathophysiology

基　　金：安徽省教育厅自然科学研究基金重点项目(No.KJ2011 A266);活性生物大分子研究安徽省重点实验室资助项目(科基[2012]126号)

摘　　要：目的:探讨核因子κB(NF-κB)和活化蛋白1(AP-1)信号通路在尖吻蝮蛇毒蛋白C激活剂(PCA)抑制脂多糖(LPS)诱导的人脐静脉内皮细胞(HUVECs)组织因子(TF)表达中的作用。方法:MTT法检测HUVECs活力,免疫组化法检测肿瘤坏死因子受体相关因子6(TRAF6)蛋白在细胞中的分布,Western blot法检测细胞内NF-κB p65、TF、c-Fos和c-Jun蛋白的表达,qPCR法测定TF mRNA在HUVECs的表达,ELISA法检测细胞培养上清中TF的含量。结果:与对照组比较,LPS组的细胞活力明显下降(P<0.01),胞质内出现明显的黄染颗粒,胞质染色加深,TRAF6平均吸光度值升高(P<0.01),NF-κB p65、c-Jun和c-Fos的蛋白表达均明显增加(P<0.01),TF mRNA和蛋白表达亦明显增加(P<0.01);PCA+LPS组的细胞活力较LPS组升高(P<0.05),细胞形态正常,胞质黄染颗粒不明显,TRAF6平均吸光度明显小于LPS组(P<0.01),NF-κB p65、c-Fos和c-Jun 3种蛋白表达则明显降低(P<0.01),TF mRNA及蛋白亦表达减少(P<0.01)。结论:PCA可明显减轻LPS引起的HUVECs损伤,其机制可能是通过降低TRAF6、NF-κB及AP-1核因子的活化进而减少组织因子的释放来实现的。AIM： To explore the role of nuclear factor-κB（ NF-κB） and activator protein-1（ AP-1） signaling pathway in the inhibitory effects of Agkistrodon acutivirus protein C activator（ PCA） on lipopolysaccharide（ LPS）-induced tissue factor（ TF） expression in human umbilical vein endothelial cells（ HUVECs）. METHODS： The viability of the HUVECs was measured by MTT assay. The protein distribution of tumor necrosis factor-associated factor 6（ TRAF6） in the cells was detected by immunohistochemical staining. The protein expression of NF-κB p65,TF,c-Fos and c-Jun was determined by Western blot. The mRNA expression of TF in the HUVECs was detected by qPCR. The content of TF in the medium of each group was measured by ELISA. RESULTS： Compared with the control group,the viability of the HUVECs in LPS group decreased significantly（ P〈0. 01）,obvious yellow dye particles appeared in the cytoplasm,cytoplasmic staining deepened,and the average absorbance of TRAF6 was increased（ P〈0. 01）. The protein expression of NF-κB p65,c-Jun and c-Fos were significantly increased（ P〈0. 01）. The expression of TF at mRNA and protein levels were significantly increased（ P〈0. 01）. Compared with the LPS group,the cell viability in PCA ＋ LPS group was slightly increased（ P〈0. 05）,the cell morphology was normal,cytoplasmic yellow dye particles were not obvious,and the average absorbance of TRAF6 was significantly lower than that in LPS group（ P〈0. 01）. The protein expression of NF-κB,c-Jun and c-Fos was significantly decreased（ P〈0. 01）,and the expression of TF at mRNA and protein levels were decreased（ P〈0. 01）. CONCLUSION： PCA significantly reduces the damage of HUVECs induced by LPS. The mechanism may be achieved by reducing the activation of TRAF6,NF-κB and AP-1 nuclear transcription factors,thereby reducing the release of tissue factor.

关 键 词：尖吻腹蛇毒蛋白C激活剂 人脐静脉血管内皮细胞 组织因子 核因子ΚB 活化蛋白1 

分 类 号：R363.2[医药卫生—病理学] R996.3[医药卫生—基础医学]