微炎症状态促进血液透析患者血清体外诱导血管新生研究

Microinflammation contributes to angiogenesis induced by serum in vitro in patients undergoing haemodialysis

目的探讨微炎症状态促进维持性血液透析患者血清体外诱导血管新生的作用,为研究透析患者临床并发症的产生机理提供依据。方法免疫速率散射比浊法及ELISA法检测血液透析患者血清超敏C-反应蛋白(hs-CRP)、肿瘤坏死因子-α(TNF-α)及白细胞介素-6(IL-6)水平,根据hs-CRP、TNF-α及IL-6检测结果,入选对象分为微炎症组和无炎症组,每组各包含9例患者,人脐静脉内皮细胞分别培养于10%微炎症组血清培养基和10%无炎症组血清培养基,光学显微镜下观察血管样结构形成,MTT法检测细胞增殖,Transwell小室测定细胞迁徙能力,Western-blot法检测p-p38蛋白表达。结果微炎症状组患者hs-CRP(7.18±0.66)mg/L、TNF-α(63.60±7.06)pg/ml及IL-6(69.77±6.96)pg/ml水平高于无炎症组,差异有统计学意义,微炎症组内皮细胞培养30h有血管样结构形成,MTT平均吸光度A值1.073±0.046升高,Transwell小室迁移过滤膜细胞数增多,10h时微炎症组内皮细胞p-p38表达增加0.268±0.023,与无炎症组比较差异均有统计学意义(P<0.05)。结论微炎症状态通过增加内皮细胞p-p38表达促进维持性血液透析患者血清在体外诱导血管新生。

OBJECTIVE To explore the contribution of microinflammation to angiogenesis induced by serum in vitro in patients receiving haemodialysis so as to provide reference for analysis on pathogenesis of complications in patients receiving haemodialysis.METHODS The serum levels of TNF-αand IL-6were determined by ELISA and the serum level of hs-CRP was investigated by immunonephelometric method.The experimental subjects were divided into the microinflammation group and the non-inflammation group with 9cases in each group.HUVECs were cultured in the 10%serum medium in the microinflammation group and the 10%serum medium in the non-inflammation group.The vessel-like structure was observed under optical microscope,cell proliferation was examined by MTT method,cell migration was measured in Transwell chamber,the p-p38 expression was evaluated by westernblot.RESULTS The serum levels of hs-CRP,TNF-αand IL-6in the microflammation group were（7.18±0.66）mg/L,（63.60±7.06）pg/ml and（69.77±6.96）pg/ml,significantly higher than those in the non-inflammation group.There were vessel-like structure formed after HUVECs were exposed to serum medium for 30 hours in the microflammation group,the average absorbance value A was increased to 1.073±0.046,the migration cells in Transwell chamber became more and the expression of p-p38 at 10hwas increased to 0.268±0.023 in the microinflammation group,the difference was significant compared with the non-inflammation group（P〈0.05）.CONCLUSION Microinflammation contributes to angiogenesis induced by serum in vitro in patients receiving haemodialysis by increasing p-p38 expression of endothelial cells.