摘要
目的探讨白蛋白导致肾小管上皮细胞NLRP3炎性体激活的机制。方法采用免疫组化法检测30例不同蛋白尿水平的膜性肾病患者肾组织中组织蛋白酶B(Cathepsin B)的表达;用BSA刺激HK-2细胞,再用高浓度KCl、Cathepsin B抑制剂CA074 Me及活性氧(ROS)抑制剂二苯基氯化碘(diphenyliodonium chloride,DPI)作用于HK-2细胞,应用Western blot、实时荧光定量PCR法检测IL-1β和IL-18生成的变化。结果大量蛋白尿的膜性肾病患者肾小管上皮细胞中Cathepsin B的表达明显高于低蛋白尿的患者(P<0.05);CA 074 Me组、DPI组的IL-1β和IL-18蛋白和mRNA生成明显减少(P<0.05),而高浓度KCl组无明显变化(P>0.05)。结论白蛋白引起的肾小管上皮细胞NLRP3炎性体激活,可能与Cathepsin B释放和ROS生成有关,而与钾离子外流无关。
Purpose To study the mechanism of NLRP3 inflammasome activation caused by albumin in renal tubulointer- stitial cells. Methods Cathepsin B was detected by immuno- histochemistry in renal biopsy tissue of 30 membranous nephrop- athy patients which had different levels of proteinuria. HK-2 cells were stimulated by albumin, and then were treated by high concentration KCI, CA 074 Me and DPI, which was Cathepsin B inhibitor and ROS inhibitor. Finally, IL-lβ and IL-18 were de- tected by Western blot and real time PCR, respectively. Results The expression of Cathepsin B in tubulointerstitial cells wassignificantly higher in patients with severe proteinuria than that in patients with mild proteinuria （ P 〈 0. 05 ）. CA 074 Me and DPI significantly reduced IL-1β and IL-18 secretion in HK-2 cells stimulated by albumin （P 〈 0. 05）, but high concentration KC1 did not result in this change （ P 〉 0. 05 ）. Conclusion NLRP3 inflammasome is activated via Cathepsin B release and increases ROS production caused by proteinuria, but not via K ＋ efflux.
出处
《临床与实验病理学杂志》
CSCD
北大核心
2017年第12期1341-1345,共5页
Chinese Journal of Clinical and Experimental Pathology
基金
国家自然科学基金重点项目(81130010)
