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目的:探讨终末期肾病患者血清对内皮细胞凋亡的影响及黄芪总黄酮的干预作用。方法:收集22例健康志愿者和25例终末期肾病规律血液透析患者的血清,在体外模拟终末期肾病患者内环境,以人脐静脉血管内皮细胞(human umbilical vein endothelial cells,HUVECs)为对象,分为空白对照组(10%胎牛血清),正常对照组(10%健康人血清),终末期肾病组(10%终末期肾病患者血清),低剂量(TFA1),中剂量(TFA2),高剂量(TFA3)组(在终末期肾病组基础上分别加入0.5,1.0,2.0 g·L-1黄芪总黄酮),培养24 h后光镜下观察血管内皮细胞形态,MTT法检测细胞生存率,免疫组化法检测C/EBP同源蛋白(CHOP)的表达,ELISA法检测4-羟基壬烯醛(4-HNE)的含量,TUNEL法检测细胞凋亡。结果:与正常对照组相比,终末期肾病患者组细胞生存率降低(P<0.01),CHOP和4-HNE升高(P<0.05),凋亡细胞数增高(P<0.01)。黄芪总黄酮干预后,各组HUVECs生存率提高(P<0.05);CHOP的表达降低(P<0.05),中剂量剂量组优于低剂量组(P<0.01),但高剂量组与中剂量组差异无统计学意义;4-HNE含量降低(P<0.05),细胞凋亡减少(P<0.05),呈剂量依赖性(P<0.05)。相关性分析显示:CHOP与4-HNE的含量呈正相关(P<0.01)。结论:终末期肾病患者血清可诱导HUNFCs凋亡,其机制部分与内质网应激诱导的细胞凋亡通路相关,黄芪总黄酮可通过减轻内质网应激从而抑制终末期肾病患者血清诱导的内皮细胞凋亡。
Objective: To investigate the effect of serum of patients with end stage renal disease on the apoptosis of endothelial cells and the intervention of total flavonoids of Astragalus. Methods: Serum samples from 22 healthy volunteers and 25 patients with end-stage renal disease undergoing hemodialysis were collected and used to simulate the internal environment of end-stage renal disease patients in vitro. Human umbilical vein endothelial cells (HUVECs) (10% fetal bovine serum), normal control group (10% healthy human serum), end-stage renal disease group (10% serum of end-stage renal disease patients), low dose (TFA1), medium dose High-dose (TFA3) group (in the end-stage renal disease group were added on the basis of 0.5,1.0,2.0 g · L-1 total flavonoids of Astragalus), cultured for 24 h after light microscopy vascular endothelial cell morphology, MTT assay cell viability The expression of C / EBP homology protein (CHOP) was detected by immunohistochemistry, the content of 4-hydroxynonenal (4-HNE) was detected by ELISA, and the apoptosis was detected by TUNEL. Results: Compared with the normal control group, the survival rate of the patients with end stage renal disease was decreased (P <0.01), CHOP and 4-HNE were increased (P <0.05), and the number of apoptotic cells was increased (P <0.01). After intervention of total flavonoids of Astragalus, the survival rate of HUVECs in each group increased (P <0.05), the expression of CHOP decreased (P <0.05), the middle dose group was better than the low dose group (P <0.01) There was no significant difference between the two groups (P <0.05). The 4-HNE content was decreased (P <0.05) and the apoptosis was decreased (P <0.05). Correlation analysis showed that: CHOP and 4-HNE content was positively correlated (P <0.01). Conclusion: The serum of patients with end stage renal disease can induce the apoptosis of HUNFCs. The mechanism is partly related to the apoptosis pathway induced by endoplasmic reticulum stress. The total flavonoids of astragalus can inhibit the serum induced by end-stage renal disease by reducing endoplasmic reticulum stress Endothelial cell apoptosis.