维持性血液透析患者血清和透析液对脂肪细胞凋亡的影响

目的观察维持性血液透析（HD）患者血清及其与常规透析液（CD）或高纯透析液（HPD）联合干预对脂肪细胞凋亡的影响。方法采集20例HD患者（CD和HPD治疗各10例）血样并分离血清,ELISA法检测血清肿瘤坏死因子α（TNF-α）水平。分别以HD患者血清（HDPS组）、HD患者血清联合CD（HDPS＋CD组）和HD患者血清联合HPD（HDPS＋HPD组）对经体外诱导分化成熟的3T3-L1脂肪细胞进行干预,48 h后收集细胞。Hoechst 33258染色荧光显微镜观察细胞形态学改变;Annexin-V-FITC/PI双标记流式细胞术检测细胞凋亡。PCR检测CD和HPD中细菌DNA片段表达。结果 ELISA法检测结果显示,与HPD治疗HD患者比较,CD治疗HD患者血清TNF-α水平显著升高（P〈0.05）。在HDPS组和HDPS＋CD组,荧光显微镜观察可见呈典型凋亡形态学改变的细胞增多,两组细胞凋亡率均显著高于HDPS＋HPD组（P〈0.05）。PCR检测显示,CD和HPD中细菌DNA表达分别为902.79±60.57和454.87±32.22,两者差异有统计学意义（P〈0.05）。结论 HDPS可诱导3T3-L1脂肪细胞凋亡,可能与患者血清TNF-α水平升高有关。与HD患者血清联合CD干预比较,联合HPD干预诱导的脂肪细胞凋亡率较低且HPD中细菌DNA表达较少。

Effects of serum of patients with maintenance hemodialysis and dialysates on apoptosis of adipocytes

Objective To investigate the effects of serum of patients with maintenance hemodialysis (HD), conventional dialysate (CD) and high purity dialysate (HPD) on apoptosis of adipocytes. Methods Blood samples of 20 patients with HD (ten were treated with CD and ten were treated with HPD) were collected, and serum was isolated for detection of tumor necrosis factor-α (TNF-α) by ELISA. Mature 3T3-L1 adipocytes induced in vitro were treated with hemodialysis patient serum (HDPS group), HDPS combined with CD (HDPS+CD group) and HDPS combined with HPD (HDPS+HPD group), respectively, and cells were harvested 48 h later. Morphological changes were observed by Hoechst33258 fluorescence staining, and cell apoptosis was detected by Annexin-V-FITC/PI double staining. Bacterial DNA from CD and HPD was assayed by PCR. Results It was revealed by ELISA that the serum level of TNF-α from patients with HD treated by CD was significantly higher than that from patients with HD treated by HPD (P<0.05). Typical apoptotic morphology changes were observed by fluorescence microscopy in HDPS group and HDPS+CD group, and the apoptosis rates in these two groups were significantly higher than that in HDPS+HPD group (P<0.05). PCR revealed that there were significant differences between the expression of bacterial DNA in CD and that in HPD (902.79±60.57 vs 454.87±32.22)(P<0.05). Conclusion HDPS may induce apoptosis of 3T3-L1 adipocytes, which may be related to the increased serum level of TNF-α. Compared with intervention by HDPS and CD, intervention by HDPS and HPD may yield lower apoptosis rate of adipocytes, and there may be lower expression of bacterial DNA in HPD.