尿毒症患者红细胞生成抑制因子的实验研究

The study of erythropoiesis inhibitors in patients with uremia

目的本文采用甲基纤维素细胞培养的方法,分析尿毒症患者血清及超滤液对小鼠骨髓红系造血祖细胞集落形成的影响,初步探讨红细胞生成抑制因子的存在、其相对分子质量大小及高通量透析器与普通低通量透析器对其清除的不同影响。方法选择北京朝阳医院肾内科12例尿毒症肾性贫血未透析患者为实验组,另选正常健康体检者12例为正常对照组,将实验组、正常对照组血清及应用F80透析器对实验组患者采用单纯超滤的方法所得超滤液(A组)、并将上述超滤液再用F6透析器进行循环超滤所得的超滤剩余液(B组)及F6超滤液(C组)分别稀释成不同终浓度,采用甲基纤维素细胞培养的方法与小鼠骨髓细胞进行培养,对比不同浓度血清组红系集落形成单位(CFU-E)、红系爆式形成单位(BFU-E)的集落数,A、B、C3组超滤液在相同浓度时CFU-E、BFU-E集落的数量。结果在实验组中,与血清浓度为0的浓度组相比,与各浓度组血清共同培养的小鼠骨髓CFU-E和BFU-E的集落数均明显减少,且随血清浓度的增加,小鼠骨髓CFU-E和BFU-E的集落数逐渐减少,各浓度组间差异有显著性(P<0.05);在A、B、C3组的各组内,与超滤液浓度为0的浓度组相比,与各浓度组超滤液共同培养的小鼠骨髓CFU-E和BFU-E的集落数均明显减少,且随超滤液浓度的增加小鼠骨髓CFU-E和BFU-E的集落数逐渐减少,各浓度组间差异有显著性(P<0.05);相同浓度时,与A组滤过液共同培养的小鼠骨髓CFU-E和BFU-E的集落数较B组、C组均明显减少;B组、C组两组之间差异无显著性(P>0.05)。结论尿毒症患者血中存在红细胞生成抑制因子,其对红系造血祖细胞的抑制作用具有剂量依赖性;红细胞生成抑制因子既可有大、中分子物质也有小分子物质;高通量透析器较普通低通量透析器能更有效清除红细胞生成抑制因子。

Objective To investigate the existence of the erythropoiesis inhibitors（EI） and the different effects of high-flux dialyzer and low-flux dialyzer on the clearance of EI. And to observe the effects of uremic patients sera and the ultrafiltrate from FSO and F6 dialyzer on CFU-E and BFU-E colony groewth of normal mice bone marrow. Methods The present study included twelve pre-dialysis ESRD patients with anemia. The sera from the uremic patients wer% added to CFU-E and BFU-E culture of normal mice bone marrow in the concentrations ranging from 1.25% to 5%.In vitro CFU-Eand BFU-E growth in the presence of sera from ESRD patients was compared with those in the presence of seva from normal human subjects. The ultrafiltrate by FSO dialyzers was defined as group A. The ultrafiltrate of group A was reeireulated and filtrated by F6 dialyzers, the remained ultrafiltrate was defined as group B, the ultrfiltrate from F6 dialyzers was as group C. The ultrafiltrate of the three groups were added to CFU-E and BFU-E culture in the concentrations ranging from 1.25% to 5%. In vitro CFU-E and BFU-E growth in the presence of ultrafiltrate of above three groups was compared with the CFU-E and BFU-E growth cultured with normal control＇s sera. Results There was no significant inhibitory effect on the CFU-E and BFU-E growth of mice bone marrow cultured in vitro with the sera of normal subjects. The inhibitory effects of ultrafiltrate in different concentrations ranging from 1.25% to 5% on CFU-E and BFU-e colony growth were dose dependent. The ultrafiltrate of the ESRD patients had inhibitory effect on in vitro CFU-E and BFU-E growth of mice bone marrow.At the same ultrafiltrate of group A had more severe inhibitory effects on the growth of CFU-E and BFU-E than those of group B and group C （P〈 0.05）. Conclusion The erythropoiesis inhibitors exist in uremic sera which has a dose-dependent inhibition effects on the CFU-E and BFU-E growth of normal mice bone marrow cells. They may be low molecular weight and medium-to-large molecular weight uremic toxins. High-flux dialyzer are more effective in clearing the erythropoiesis inhibitors than low flux dialvzer.