Wu C C, Liao T N, Lu K C, Chen J S, Chu P, Lin S H, Chuang C H, Lin Y F
Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei, Taiwan.
Clin Nephrol. 2005 Sep;64(3):198-204. doi: 10.5414/cnp64198.
There is an increased rate of apoptosis of peripheral blood mononuclear cells (PBMCs) in patients undergoing hemodialysis (HD), but little is known about how different dialysis membranes may contribute to the process. We, therefore, studied the influence of two different dialysis membranes on apoptotic markers during HD.
8 healthy controls and 8 patients on regular HD 3 times per week were enrolled in this cross-controlled study. Patients received HD using polysulfone and then regenerated cellulose dialysis membranes for one week each, sequentially. Serum was collected for C-reactive protein (CRP) detection; flow cytometry with dual antibody staining was used to measure the apoptotic markers Fas (CD95), FasL (CD 178) and TNF-R2 (CD120b) in T cells (CD3+), B cells (CD19+), and monocytes (CD14+) at 0, 15, 120 and 240 min after starting HD. We also measured total leukocyte numbers and differential white cell counts.
Hemodialysis patients revealed lymphocytopenia, monocytopenia, higher CRP levels and higher Fas and TNF-R2 expression on lymphocytes and monocytes at baseline when compared with normal controls. Leukocyte numbers, including neutrophils, lymphocytes and monocytes, dropped significantly after 15 min of dialysis. There were no significant differences in Fas levels during hemodialysis on T and B lymphocytes or on monocytes. T lymphocyte FasL (CD 178) levels remained unchanged throughout the process. There was a significantly lower overall level of CD120b at 15 min of HD, whereas this marker was higher on monocytes after dialysis. There were no significant differences in the levels of apoptotic markers between the two membranes.
Our results suggest that uremia itself contributes to PBMC apoptosis. The two different dialysis membranes used in this study did not influence apoptotic markers on PBMCs significantly, but increased TNF-R2 expression on monocytes during a single dialysis session.
血液透析(HD)患者外周血单个核细胞(PBMC)的凋亡率增加,但对于不同的透析膜如何影响这一过程知之甚少。因此,我们研究了两种不同透析膜在血液透析过程中对凋亡标志物的影响。
本交叉对照研究纳入了8名健康对照者和8名每周规律进行3次血液透析的患者。患者依次使用聚砜膜和再生纤维素透析膜进行一周的血液透析。采集血清检测C反应蛋白(CRP);采用双抗体染色流式细胞术检测血液透析开始后0、15、120和240分钟时T细胞(CD3+)、B细胞(CD19+)和单核细胞(CD14+)中的凋亡标志物Fas(CD95)、FasL(CD178)和TNF-R2(CD120b)。我们还检测了白细胞总数和白细胞分类计数。
与正常对照组相比,血液透析患者在基线时表现为淋巴细胞减少、单核细胞减少、CRP水平升高,淋巴细胞和单核细胞上的Fas和TNF-R2表达更高。透析15分钟后,包括中性粒细胞、淋巴细胞和单核细胞在内的白细胞数量显著下降。血液透析过程中,T和B淋巴细胞或单核细胞上的Fas水平无显著差异。整个过程中T淋巴细胞FasL(CD178)水平保持不变。血液透析15分钟时CD120b的总体水平显著降低,而透析后该标志物在单核细胞上的水平更高。两种透析膜之间凋亡标志物水平无显著差异。
我们的结果表明,尿毒症本身会导致PBMC凋亡。本研究中使用的两种不同透析膜对PBMC上的凋亡标志物没有显著影响,但在单次透析过程中会增加单核细胞上TNF-R2的表达。