Zhang Yu-Xin, Wan Hui, Shan Guan-Yue, Cheng Jun-Ya, Gao Zhi-Cheng, Liu Yi-Ying, Shi Wen-Na, Sun Zi-Jun, Li Hai-Jun
Department of Transplantation Immunology, Institute of Translational Medicine, The First Hospital of Jilin University, Changchun, Jilin Province, China.
Department of Bioengineering, Pharmacy School of Jilin University, Changchun, Jilin Province, China.
J Ginseng Res. 2025 Jul;49(4):426-437. doi: 10.1016/j.jgr.2025.03.009. Epub 2025 Mar 25.
Hyperuricemia nephropathy (HN) is a form of renal injury caused by hyperuricemia, which can progress to chronic kidney disease (CKD) and end-stage renal disease (ESRD). Ginsenoside Rg5, a major bioactive compound isolated from Panax ginseng, is recognized for its notable effects, including anti-inflammatory, antioxidant, and anticancer activities.
The toxic doses of MSU crystals and Rg5-induced HK-2 cell damage were assessed using the CCK-8 assay and quantifying oxidative stress markers (MDA, GSH, SOD). Intracellular stress was evaluated with JC-1 and DCFH-DA probes. Bioinformatics analysis identified NOX1, TLR4, and Bcl-2 as potential targets. The protein expression associated with stress, inflammation, pyroptosis, and apoptosis in HK-2 cells was evaluated through a combination of Western blotting, ELISA, flow cytometry, immunofluorescence, and overexpression methods. An HN mice model was established through administration of YE and adenine, and the effects of Rg5 were evaluated. The in vivo mechanisms were further verified.
Rg5 reduced serum uric acid, BUN, ADH, and creatinine levels in MSU crystals-stimulated HK-2 cells and hyperuricemic mice, alleviating renal damage. Rg5 inhibited NOX1 and suppressed the TLR4 pathway, reducing oxidative stress, inflammation, pyroptosis, and apoptosis. NOX1 overexpression reversed the effects of Rg5, while TLR4 overexpression had no effect. Rg5's efficacy was similar to NOX1 inhibitor ML171.
These results indicate that Rg5 can modulate the TLR4 and BCL-2 pathways by inhibiting NOX1, thereby alleviating oxidative stress, inflammation, pyroptosis, and apoptosis in HN, highlighting its potential as a therapeutic approach for controlling HN.
高尿酸血症肾病（HN）是一种由高尿酸血症引起的肾损伤形式，可进展为慢性肾脏病（CKD）和终末期肾病（ESRD）。人参皂苷Rg5是从人参中分离出的一种主要生物活性化合物，因其具有抗炎、抗氧化和抗癌等显著作用而受到认可。
使用CCK-8法并定量氧化应激标志物（丙二醛、谷胱甘肽、超氧化物歧化酶）评估单钠尿酸盐（MSU）晶体和Rg5诱导的HK-2细胞损伤的毒性剂量。用JC-1和二氯荧光素二乙酸酯（DCFH-DA）探针评估细胞内应激。生物信息学分析确定NADPH氧化酶1（NOX1）、Toll样受体4（TLR4）和B细胞淋巴瘤-2（Bcl-2）为潜在靶点。通过蛋白质免疫印迹、酶联免疫吸附测定、流式细胞术、免疫荧光和过表达方法相结合，评估HK-2细胞中与应激、炎症、细胞焦亡和凋亡相关的蛋白质表达。通过给予酵母提取物（YE）和腺嘌呤建立HN小鼠模型，并评估Rg5的作用。进一步验证体内机制。
Rg5降低了MSU晶体刺激的HK-2细胞和高尿酸血症小鼠的血清尿酸、血尿素氮、抗利尿激素和肌酐水平，减轻了肾损伤。Rg5抑制NOX1并抑制TLR4通路，减少氧化应激、炎症、细胞焦亡和凋亡。NOX1过表达逆转了Rg5的作用，而TLR4过表达则无影响。Rg5的疗效与NOX1抑制剂ML171相似。
这些结果表明，Rg5可通过抑制NOX1调节TLR4和BCL-2通路，从而减轻HN中的氧化应激、炎症、细胞焦亡和凋亡，突出了其作为控制HN的治疗方法的潜力。
