背景:糖尿病肾病(DN)是一种危及生命的肾脏疾病,需要紧急治疗。Wogonin在DN中具有肾脏保护作用。本研究旨在探讨汉黄芩素调控高糖诱导的肾细胞损伤的机制。
方法:糖尿病小鼠(db/db),对照db/m小鼠,和正常葡萄糖(NG)-或HG处理的人小管上皮细胞(HK-2)用于评估细胞因子信号抑制因子3(SOCS3)的水平,Toll样受体4(TLR4),炎症和纤维化。慢病毒用于调节SOCS3和TLR4的表达。在db/db小鼠中口服wogonin(10mg/kg)或媒介物后,组织学形态,血糖,尿蛋白,血清肌酐值(Scr),血尿素氮(BUN),超氧化物歧化酶(SOD),谷胱甘肽(GSH),和活性氧(ROS)进行了评估。RT-qPCR和Western印迹评估炎症和纤维化相关分子。
结果:HG暴露导致高血糖,严重的肾损伤,高血清Src和BUN,低SOD和GSH,增加ROS。HG下调SOCS3,但上调TLR4和JAK/STAT,纤维化,和与炎症相关的蛋白质.Wogonin通过降低细胞因子减轻HG诱导的肾损伤,ROS,Src,和MDA和增加SOD和GSH。同时,Wogonin在HG条件下上调SOCS3并下调TLR4。Wogonin诱导的SOCS3过表达直接降低TLR4水平,减轻JAK/STAT信号通路相关的炎症和纤维化,但SOCS3敲除显著拮抗汉黄芩素的保护作用。然而,TLR4敲除减少SOCS3敲除诱导的肾损伤。
结论:Wogonin通过上调SOCS3抑制TLR4和JAK/STAT通路减轻肾脏炎症和纤维化。
BACKGROUND: Diabetic nephropathy (DN) is a life-threatening renal disease and needs urgent therapies. Wogonin is renoprotective in DN. This study aimed to explore the mechanism of how wogonin regulated high glucose (HG)-induced renal cell injury.
METHODS: Diabetic mice (db/db), control db/m mice, and normal glucose (NG)- or HG-treated human tubule epithelial cells (HK-2) were used to evaluate the levels of suppressor of cytokine signaling 3 (SOCS3), Toll-like receptor 4 (TLR4), inflammation and fibrosis. Lentivirus was used to regulate SOCS3 and TLR4 expressions. After oral gavage of wogonin (10 mg/kg) or vehicle in db/db mice, histological morphologies, blood glucose, urinary protein, serum creatinine values (Scr), blood urea nitrogen (BUN), superoxide dismutase (SOD), glutathione (GSH), and reactive oxygen species (ROS) were assessed. RT-qPCR and Western blot evaluated inflammation and fibrosis-related molecules.
RESULTS: HG exposure induced high blood glucose, severe renal injuries, high serumal Src and BUN, low SOD and GSH, and increased ROS. HG downregulated SOCS3 but upregulated TLR4 and JAK/STAT, fibrosis, and inflammasome-related proteins. Wogonin alleviated HG-induced renal injuries by decreasing cytokines, ROS, Src, and MDA and increasing SOD and GSH. Meanwhile, wogonin upregulated SOCS3 and downregulated TLR4 under HG conditions. Wogonin-induced SOCS3 overexpression directly decreased TLR4 levels and attenuated JAK/STAT signaling pathway-related inflammation and fibrosis, but SOCS3 knockdown significantly antagonized the protective effects of wogonin. However, TLR4 knockdown diminished SOCS3 knockdown-induced renal injuries.
CONCLUSIONS: Wogonin attenuates renal inflammation and fibrosis by upregulating SOCS3 to inhibit TLR4 and JAK/STAT pathway.