CAJ | 학술논문

目的观察白蛋白对肾小管上皮细胞中NLRP3炎性体及其下游炎性因子IL-1β和IL-18表达的影响.方法选取30例不同蛋白尿水平的系膜增生性肾小球肾炎(MsPGN)患者肾活检组织,分别用PAS和Masson染色观察肾小管间质损伤和炎性细胞的浸润情况;用免疫组化的方法检测其肾小管上皮细胞中NLRP3、caspase-1、IL-1β和IL-18的表达情况,以及间质中浸润的不同炎性细胞.体外实验用高浓度的牛血清白蛋白(BSA,20 g/L)刺激HK-2细胞,采用Western印迹和实时荧光定量PCR方法观察NLRP3、caspase-1、IL-1β和IL-18的蛋白和mRNA表达水平的变化.结果高蛋白尿组MsPGN患者的肾小管损伤和炎性细胞浸润程度明显重于低蛋白尿组(均P＜ 0.05),肾小管上皮细胞中NLRP3、caspase-1、IL-1β和IL-18的表达也明显高于低蛋白尿组(均P＜ 0.05),且IL-1β和IL-18的表达量与24h尿蛋白量呈正相关(r=0.836,P＜0.05;r=0.901,P＜0.05).体外BSA刺激HK-2细胞24 h后,NLRP3、caspase-1、IL-1β和IL-18蛋白和mRNA水平均明显高于对照组(均P＜0.05).结论白蛋白可引起肾小管上皮细胞中NLRP3炎性体的激活,这可能是蛋白尿导致肾小管间质损伤和炎性反应的机制.
목적 관찰백단백대신소관상피세포중NLRP3염성체급기하유염성인자IL-1β화IL-18표체적영향.방법 선취30례불동단백뇨수평적계막증생성신소구신염(MsPGN)환자신활검조직,분별용PAS화Masson염색관찰신소관간질손상화염성세포적침윤정황;용면역조화적방법검측기신소관상피세포중NLRP3、caspase-1、IL-1β화IL-18적표체정황,이급간질중침윤적불동염성세포.체외실험용고농도적우혈청백단백(BSA,20 g/L)자격HK-2세포,채용Western인적화실시형광정량PCR방법관찰NLRP3、caspase-1、IL-1β화IL-18적단백화mRNA표체수평적변화.결과 고단백뇨조MsPGN환자적신소관손상화염성세포침윤정도명현중우저단백뇨조(균P＜ 0.05),신소관상피세포중NLRP3、caspase-1、IL-1β화IL-18적표체야명현고우저단백뇨조(균P＜ 0.05),차IL-1β화IL-18적표체량여24h뇨단백량정정상관(r=0.836,P＜0.05;r=0.901,P＜0.05).체외BSA자격HK-2세포24 h후,NLRP3、caspase-1、IL-1β화IL-18단백화mRNA수평균명현고우대조조(균P＜0.05).결론 백단백가인기신소관상피세포중NLRP3염성체적격활,저가능시단백뇨도치신소관간질손상화염성반응적궤제.
Objective To investigate the effect of albumin on expression of NLRP3 inflammasome and its downstream cytokines IL-1β and IL-18 in tubular epithelial cells.Methods Thirty mesangioproliferative glomerulonephritis (MsPGN) patients with different levels of proteinuria were selected, and their renal biopsy samples were stained by PAS and Masson to observe tubular epithelial cells injury and inflammatory cells infiltration.NLRP3, caspase-1, IL-1β and IL-18, as well as different inflammatory cells, were detected by immunohistostaining.In vitro, Western blotting and real-time PCR were employed to detect NLRP3, caspase-1, IL-1β and IL-18 protein and mRNA in HK-2 cells stimulated by bovine serum albumin (BSA) (20 g/L).Results In MsPGN patients with high levels of proteinuria, there were obvious renal tubular epithelial cell injury and inflammatory cells infiltration (all P ＜ 0.05), and the expressions of NLRP3, caspase-1, IL-1β and IL-18 were up-regulated compared to patients with low levels of proteinuria (all P ＜ 0.05).Furthermore, IL-1β and IL-18expressions were positively correlated with the degree of proteinuria (r=0.836, P ＜ 0.05;r=0.901, P ＜0.05).NLRP3, caspase-1, IL-1β and IL-18 protein and mRNA were significantly increased in HK-2cells stimulated by BSA compared to the control group (all P ＜ 0.05).Conclusions Albumin is able to induce NLRP3 inflammasome activation in tubular epithelial cells, which may be the mechanism of tubulointerstitial injury and inflammation caused by proteinuria.