肌酐产物导致红细胞溶血机理探讨(1)
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【摘要】目的：研究肌酐产物对红细胞的毒性作用。方法：用液相色谱仪分析尿毒症患者肠道细菌分解肌酐的产物。用分解肌酐能力最强的肺炎克雷伯菌与肌酐共同培养，取其培养液与健康红细胞孵化，观察溶血情况；并分别加入葡萄糖、ATP(三磷酸腺苷)、GSH(谷胱甘肽)，了解以上物质对溶血的纠正情况；并测定红细胞内GSH的浓度。结果：肠道细菌分解肌酐产生甲胺。肌酐组溶血度1.52%，肌酐产物组3.52%，后者较前者的毒性强，加重红细胞溶血(P<0.05)；加入葡萄糖、ATP溶血不纠正，加入GSH明显纠正(P<0.001)；GSH组溶血度0.252%，正常对照组0.192%，二者差异无显著性。肌酐组红细胞内GSH含量2152.98μmol/L，肌酐产物组1047.96μmol/L，后者红细胞内GSH含量明显降低(P<0.001)。 结论：肌酐产物导致红细胞溶血，红细胞内GSH含量下降，补充GSH可纠正溶血。
【关键词】肌酐产物;红细胞;溶血;GSH
【中图分类号】R331.1【文献标识码】B【文章编号】1008-6455(2011)08-0385-03
The effection and mechanismthat products metabolism of creatinine treated erythrocytes
Hu Baiying
【Abstract】Objective: To study the toxicity that metabolite of creatinine treatederythrocytes.Methods：Metabolist of creatinine of uraemia intestinal bacteria was confirmed by HPLC. Klebsiella pneumoniae were incubated with creatinine, it’s metabolite and erythrocytes were incubated. hemolysiswas observed and GSH was determined in erythrocytes.Results：Metabolist of creatinine was methylamine, which can bring hemolysis of erythrocytes(P<0.05). Degree of hemolysis of creatinine group was 1.52% and that of metabolist of creatinine group was 3.52%. Added up glucose and ATP, the hemolysis was not reversed. Added up GSH, it was reversed (P<0.01). Concentration of GSH in erythrocytes of metabolist of creatinine group was decreased (P<0.001). Concentration of GSH of creatinine group was 2152.98μmol/L and that of metabolist of creatinine group was 1047.49μmol/L. Conclusions：The toxicity to erythrocytes of the metabolite of creatinine is stronger than that of creatinine. The toxicity could be reversed by GSH.
【Key words】ErythrocytesGSHMetabolite of creatinine
慢性肾衰酶疗法，不仅要研究酶的结构、功能，最大限度发挥其分解作用，但对分解后的产物对人体的作用的研究也是至关重要的。毒素分解的下限物质必须是无毒的，对人体组织细胞安全。因而研究产物的毒性作用，是关系酶疗法的重要内容。国内对该领域报道较少，该部分则探讨肌酐产物对红细胞的影响。
1 材料
(1)利用肌酐能力最强的尿毒症患者的肠道细菌——肺炎克雷伯菌(实验已另文发表)。
(2)肌酐。
(3)正常健康志愿者的红细胞。
2 方法
2.1 肌酐产物的分析:肌酐对照液的配制：肌酐，用双蒸水配制成0.14mg/ml。
肌酐转化为甲基胍的对照液配制：肌酐230 mg，三氯化铁270 mg，1%过氧化氢100 ml，于震荡恒温培养箱，25℃温育3小时，即得。其为肌酐、甲基胍及中间代谢产物。
肠道细菌作用于肌酐的代谢产物的配制：肌酐1200μmol/ml溶液1ml，肠道细菌1菌环，37℃恒温培养箱培养48小时，离心取上清液。
细菌将肌酐转化为甲胺的对照液配制：取分解肌酐为甲胺的土壤细菌斯氏假单胞菌(Pseudomonas stutzeri)一菌环，余操作同3。 ......
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