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纸质出版:2014
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韦杏雪,吴军,梁新,丁瑜,袁发焕.蛋白结合硫酸吲哚酚损伤红细胞膜[J].陆军军医大学学报(原第三军医大学学报),2014,36(24):2467-2470.
Wei Xingxue,Wu Jun,Liang Xin,Ding Yu,Yuan Fahuan.Albumin-bound indoxyl sulfate damages erythrocytes in vitro[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(24):2467-2470.
目的 研究蛋白结合硫酸吲哚酚对红细胞寿命的影响及作用机制。 方法 以4%白蛋白结合不同浓度的硫酸吲哚酚孵育0.4%红细胞悬液48 h
采用流式细胞仪检测红细胞体积,Annexin-V-FITC凋亡试剂盒检测红细胞表面磷脂酰丝氨酸的标记率,钙离子荧光探针(Fluo3/AM)检测红细胞内Ca
2+
浓度,酶标分析仪检测红细胞的溶血率,采用SPSS 18.0统计软件进行统计分析。 结果 4%白蛋白结合硫酸吲哚酚孵育0.4%红细胞悬液48 h后,随硫酸吲哚酚浓度逐渐增加,红细胞体积逐渐变小[(1603.33±8.50)vs(742.67±14.04),P0.05]
红细胞表面磷脂酰丝氨酸的标记率[(0.26±0.06)vs(10.32±1.64),P0.05]、红细胞内Ca
2+
浓度以及溶血率逐渐增加,与对照组相比差异有统计学意义(P0.05),呈剂量依赖性;当细胞外Ca
2+
缺乏时,硫酸吲哚酚对红细胞的这种损伤效应明显降低。 结论 4%白蛋白结合硫酸吲哚酚通过增加钙内流而损伤红细胞,促进红细胞衰亡,是尿毒症患者贫血的发生机制之一,清除蛋白结合毒素有助于改善贫血。
Objective To investigate the effect of albumin-bound indoxyl sulfate on erythrocytes and related mechanism. Methods Different concentrations of indoxyl sulfate were bound to 4% albumin. Erythrocytes with 0.4% hematocrit were prepared from normal adults. Forward scatter was used to estimate erythrocyte volume. Phosphatidylserine-exposure from annexin V binding was used to check eryptosis. Erythrocyte intracellular free calcium concentration ([Ca
2+
]
i
) was detected by Fluo3-fluorescence and assayed by flow cytometry. Hemolysis from hemoglobin release was
estimated with an enzyme-mark analyzer. Statistical analysis was made with SPSS18.0 software. Results Exposure to albumin-bound indoxyl sulfate for 48 hours significantly decreased erythrocyte volume (1603.33±8.50 vs 742.67±14.04
P0.05)
and increased eryptosis (0.26±0.06 vs 10.32±1.64
P0.05)
[Ca
2+
]
i
and hemolysis ratio in a dose-dependent manner (P0.05). The damage effect of albumin-bound indoxyl sulfate on erythrocytes was weakened in the absence of Ca
2+
in the supernatant. Conclusion Albumin-bound indoxyl sulfate induces eryptosis through calcium influx
which is an important mechanism of anemia in uremic patients. Eliminating albumin-bound indoxyl sulfate might alleviate renal anemia.
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