Li Dengren, Yu Kuipeng, Feng Feng, Zhang Yang, Bai Fang, Zhang Yimeng, Sun Nan, Fan Jiahui, Liu Lei, Yang Huimin, Yang Xiangdong
Department of Nephrology, Qilu Hospital of Shandong University, Jinan, 250012, Shandong, China.
Department of Nephrology, Qilu Hospital of Shandong University, Jinan, 250012, Shandong, China; Laboratory of Basic Medical Sciences, Qilu Hospital of Shandong University, Jinan, 250012, Shandong, China.
Biochem Biophys Res Commun. 2022 Jun 25;610:154-161. doi: 10.1016/j.bbrc.2022.04.058. Epub 2022 Apr 15.
Renal fibrosis is the ultimate presentation of chronic kidney disease, which progresses to end-stage renal disease. Hydroxychloroquine (HCQ) has been adapted for the treatment of autoimmune diseases; however, the potential mechanism underlying the role of HCQ in renal fibrosis remains unclear. C57BL/6 J mice were randomly divided into three groups (sham group, UUO group, and UUO + HCQ group (20 mg/kg)). HE and Masson staining were performed to assess kidney tissue damage and fibrosis, and western blotting was performed to assess the expression of epithelial-mesenchymal transition (EMT), extracellular matrix (ECM), PI3K/AKT, and NF-κB-related proteins. PCR and TUNEL were adopted to detect inflammatory factors and cell apoptosis. HK-2 cells treated with TGF-β1 were used for the in vitro experiments. HCQ may potentially have therapeutic effects on renal fibrosis mediated through 122 target genes, and the Kyoto Encyclopedia of Genes and Genomes pathways of these genes were enriched for PI3K/AKT signaling based on network pharmacology. UUO mice that received HCQ demonstrated significantly less tubular damage than the UUO mice. HCQ treatment additionally blunted EMT in UUO kidneys and TGF-β1-treated renal tubular epithelial cells, and alleviated ECM deposition in kidney tissue. Furthermore, HCQ treatment reduced UUO-induced inflammation and apoptosis. Mechanistically, HCQ treatment suppressed the activation of the PI3K/Akt and NF-kB pathways. This study demonstrated that HCQ ameliorated renal fibrosis by inhibiting the PI3K/AKT and NF-κB signaling pathways to attenuate inflammatory factors and the apoptotic function of renal tubular epithelial cells, thus providing renewed theoretical evidence for HCQ treatment of renal fibrosis.
肾纤维化是慢性肾脏病的最终表现形式，可进展为终末期肾病。羟氯喹（HCQ）已被用于治疗自身免疫性疾病；然而，HCQ在肾纤维化中作用的潜在机制仍不清楚。将C57BL/6 J小鼠随机分为三组（假手术组、单侧输尿管梗阻（UUO）组和UUO + HCQ组（20 mg/kg））。进行苏木精-伊红（HE）和Masson染色以评估肾组织损伤和纤维化，进行蛋白质免疫印迹法以评估上皮-间质转化（EMT）、细胞外基质（ECM）、磷脂酰肌醇-3激酶/蛋白激酶B（PI3K/AKT）和核因子-κB（NF-κB）相关蛋白的表达。采用聚合酶链反应（PCR）和末端脱氧核苷酸转移酶介导的缺口末端标记法（TUNEL）检测炎症因子和细胞凋亡。用转化生长因子-β1（TGF-β1）处理的人肾小管上皮细胞（HK-2细胞）用于体外实验。基于网络药理学，HCQ可能对通过1,22个靶基因介导的肾纤维化具有治疗作用，这些基因的京都基因与基因组百科全书（KEGG）通路在PI3K/AKT信号传导方面显著富集。接受HCQ治疗的UUO小鼠肾小管损伤明显少于UUO小鼠。HCQ治疗还减弱了UUO肾脏和TGF-β1处理的肾小管上皮细胞中的EMT，并减轻了肾组织中的ECM沉积。此外，HCQ治疗减轻了UUO诱导的炎症和凋亡。机制上，HCQ治疗抑制了PI3K/Akt和NF-κB通路的激活。本研究表明，HCQ通过抑制PI3K/AKT和NF-κB信号通路来减轻炎症因子和肾小管上皮细胞的凋亡功能，从而改善肾纤维化，为HCQ治疗肾纤维化提供了新的理论依据。
