摘要
目的:探究近端肾小管上皮细胞中肾损伤分子1(KIM-1)介导免疫球蛋白轻链(FLCs)内吞对肾小管损伤的影响及机制。方法:(1)选取4例多发性骨髓瘤肾损伤患者尿标本,采用离子交换层析法纯化FLCs;(2)体外实验构建纯化的FLCs诱导人肾脏近端肾小管上皮细胞(HK-2细胞)损伤模型,继续给予FLCs刺激,并予KIM-1抑制剂TW-37干预;体内实验向小鼠腹腔注射FLCs,14 d后建立FLCs致小鼠肾小管损伤的动物模型,并予TW-37干预。Western Blot检测HK-2细胞和小鼠肾组织KIM-1、megalin、核因子E2相关因子2(Nrf2)、葡萄糖调节蛋白78(GRP78)、α平滑肌肌动蛋白(α-SMA)及转化生长因子β(TGF-β)等水平,ELISA法检测小鼠尿白蛋白/肌酐比值、尿液KIM-1的含量,天狼星红染观察小鼠肾间质纤维化程度等。结果:(1)体外实验中FLCs刺激HK-2细胞诱导KIM-1表达增加,并与FLCs存在共定位,TW-37处理后可减少HK-2细胞中FLCs内吞,减轻FLCs刺激所致的肾小管细胞溶酶体的核周聚集,降低氧化应激及纤维化指标表达。(2)动物模型结果显示,TW-37干预可缓解FLCs所致的肾小管损伤,包括尿KIM-1水平及肾组织KIM-1表达下降,肾组织氧化应激和纤维化指标表达下降,肾组织胶原沉积减轻。结论:KIM-1可介导肾近端小管细胞内吞FLCs,抑制KIM-1有助于缓解FLCs造成的肾损伤。
Objective:To investigate effect and mechanism of kidney injury molecule-1(KIM-1)mediated immunoglobulin free light chains(FLCs)endocytosis on renal tubular injury and fibrosis in renal tubular epithelial cells.Methodology:(1)urine samples from 4 patients with multiple myeloma were selected and FLCs were purified by ion exchange chromatography;(2)the model of HK-2 cell injury was induced by FLCs extracted by the above method in vitro.FLCs stimulation was continued and KIM-1 inhibitor TW-37 was used to treat it.After continous intraperitoneal injection of FLCs for 14 days into mice in vivo,the animal model of renal tubular injury induced by FLCs was established and interfered with TW-37.The levels of KIM-1,TGF-βetc in HK-2 cells and renal tissue were detected by Western Blot.The Ratio of urinary albumin protein to creatinine and content of KIM-1 in urine were detected by ELISA method,and the degree of renal interstitial fibrosis was detected by Sirius red staining.Results:(1)in vitro,FLCs stimulated HK-2 cells to increase expression of KIM-1,which was co-localization of KIM-1 and FLCs in HK-2 cells.TW-37 treatment can reduce FLCs endocytosis in HK-2 cells,reduce perinuclear aggregation of lysosomes in renal tubular cells stimulated by FLCs,and reduce expression of oxidative stress and fibrosis indexes.The results of animal model showed that TW-37 can alleviate renal tubular injury induced by FLCs,such as decrease of KIM-1 content in urine and expression of KIM-1 in renal tissue,the decrease of oxidative stress,fibrosis index and collagen deposition in renal tissue.Conclusion:KIM-1 can mediate endocytosis of FLCs by renal proximal tubular cells,and inhibition of KIM-1 may alleviate renal injury caused by FLCs.
作者
董世惠
范芸
夏园园
杨雪
刘丰
杨帆
梁丹丹
曾彩虹
DONG Shihui;FAN Yun;XIA Yuanyuan;YANG Xue;LIU Feng;YANG Fan;LIANG Dandan;ZENG Caihong(National Clinical Research Center of Kidney Diseases,Jinling Hospital,Nanjing University School of Medicine,Nanjing 210016,China)
出处
《肾脏病与透析肾移植杂志》
CAS
CSCD
北大核心
2023年第2期121-127,共7页
Chinese Journal of Nephrology,Dialysis & Transplantation
基金
国家自然科学基金面上项目(82070793)
江苏省卫生健康科研项目(ZD2021018)。
