蛋白结合硫酸吲哚酚对单核细胞趋化功能的影响

发布时间：2018-06-06 22:54

本文选题：蛋白结合毒素 + 硫酸吲哚酚　；参考：《第三军医大学学报》2016年09期

【摘要】：目的研究蛋白结合硫酸吲哚酚(indoxyl sulfate,IS)对单核细胞趋化功能的影响及其可能机制。方法以4%人血白蛋白(human serum albumin,HSA)结合游离的IS制备蛋白结合IS,高效液相色谱法(HPLC)检测其蛋白结合率。CCK-8法检测人单核细胞(THP-1)经不同浓度的游离IS和蛋白结合IS分别作用24 h后细胞增殖能力的变化。200μmol/L游离IS和蛋白结合IS分别孵育THP-1细胞24 h后,Transwell小室法检测THP-1细胞趋化能力;ELISA法检测细胞培养上清中单核细胞趋化蛋白-1(MCP-1)浓度;流式细胞术检测THP-1细胞上CC类趋化因子受体2(CCR2)的表达水平。结果成功制备蛋白结合IS,其蛋白结合率约为90%;蛋白结合IS和游离IS均可抑制THP-1细胞增殖,且蛋白结合IS对THP-1细胞的增殖抑制作用具有浓度依赖性(P0.05);蛋白结合IS和游离IS均可增强THP-1细胞的趋化活性(P0.05);蛋白结合IS和游离IS均可使THP-1细胞的MCP-1和CCR2表达上调(P0.05)。结论蛋白结合IS同游离IS一样,可通过上调MCP-1及CCR2的表达增强单核细胞的趋化功能。
[Abstract]:Objective to study the effect of protein binding indoxyl sulfate (indoxyl sulfate) on the chemotaxis of monocytes and its possible mechanism. Methods Human serum albumin (HSA) combined with free is protein was used to bind ISs. The protein binding rate of HPLCwas determined by HPLC. CCK-8 was used to detect the binding rate of human monocytes to different concentrations of free is and protein-binding is. After 24 h incubation of THP-1 cells with 200 渭 mol/L free is and protein binding is, the chemotaxis of THP-1 cells was detected by transwell chamber assay and the concentration of monocyte chemoattractant protein-1 (MCP-1) in the supernatant was detected by Elisa. The expression of CCR2 on THP-1 cells was detected by flow cytometry. Results the protein binding rate was about 90%, and both protein binding is and free is could inhibit the proliferation of THP-1 cells. The inhibitory effect of protein-binding is on the proliferation of THP-1 cells was concentration-dependent, protein-binding is and free is could enhance the chemotaxis activity of THP-1 cells, and protein binding is and free is could up-regulate the expression of MCP-1 and CCR2 in THP-1 cells. Conclusion protein binding is can enhance the chemotaxis function of monocytes by up-regulating the expression of MCP-1 and CCR2.
【作者单位】：第三军医大学新桥医院肾内科;
【分类号】：R692

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