目的 基于高迁移率族蛋白B1（high mobility group box 1，HMGB1）-晚期糖基化末端受体（receptor for advanced glycation end products，RAGE）/Toll样受体（Toll-like receptors，TLRs）-核因子-κB（nuclear factor-κB，NF-κB）信号通路探讨红芪多糖对糖尿病肾病（diabetic kidney disease，DKD）的治疗作用。方法 db/db小鼠随机分为模型组、二甲双胍（200 mg/kg）组和红芪多糖高、中、低剂量（200、100、50 mg/kg）组，每组10只，另取10只db/m小鼠作为对照组。连续给药4周，每周测定体质量和空腹血糖。给药结束后，测定血脂、血肌酐、24 h尿蛋白、胰岛素和炎症因子水平；采用苏木素-伊红（hematoxylin-eosin，HE）染色观察肾组织病理变化；采用透射电子显微镜观察肾组织超微结构变化；采用qRT-PCR和Western blotting检测肾组织HMGB1-RAGE/TLRs-NF-κB信号通路相关基因及蛋白表达变化。采用50 mmol/L葡萄糖诱导SV40 MES-13小鼠肾小球系膜细胞凋亡，给予红芪多糖干预后，采用流式细胞仪检测细胞凋亡情况；采用Western blotting检测HMGB1-RAGE/TLRs-NF-κB信号通路相关蛋白表达变化。结果 与模型组比较，红芪多糖组小鼠体质量和空腹血糖显著降低（P＜0.001），24 h尿蛋白含量显著减少（P＜0.01、0.001），血清中胰岛素、总胆固醇、低密度脂蛋白胆固醇、血肌酐、三酰甘油水平均显著降低（P＜0.05、0.01、0.001），高密度脂蛋白胆固醇水平显著升高（P＜0.001），血清中白细胞介素-6（interleukin-6，IL-6）、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）和IL-1β水平显著降低（P＜0.05、0.01、0.001），肾脏病理损伤得到显著改善，肾组织中HMGB1-RAGE/TLRs-NF-κB信号通路相关基因及蛋白表达水平均显著降低（P＜0.05、0.01、0.001）。细胞实验结果显示，红芪多糖显著抑制高糖诱导的SV40 MES-13细胞凋亡（P＜0.05、0.001），且抑制HMGB1-RAGE/TLRs-NF-κB信号通路活化和炎症因子产生（P＜0.05、0.01、0.001）。结论 红芪多糖能够通过抑制HMGB1-RAGE/TLRs-NF-κB信号通路活化，降低炎症因子表达，从而缓解DKD。

Objective To explore the therapeutic effect of Hedysarum polybotrys polysaccharide (HPS) on diabetic kidney disease (DKD) based on high mobility group box 1 (HMGB1)-receptor for advanced glycation end products (RAGE)/Toll-like receptors (TLRs)-nuclear factor-κB (NF-κB) signaling pathway. Methods db/db mice were randomly divided into model group, metformin (200 mg/kg) group, HPS high-, medium- and low-dose (200, 100, 50 mg/kg) groups, with 10 mice in each group. An additional 10 db/m mice were used as the control group. Drugs were administered continuously for four weeks, body weight and fasting blood glucose were measured weekly. After drug administration, blood lipid, creatinine, 24 h urinary protein, insulin and inflammatory factor levels were assessed. Hematoxylin-eosin (HE) staining was used to observe renal histopathological changes. Transmission electron microscopy was employed to examine ultrastructural changes in renal tissue. qRT-PCR and Western blotting were used to detect changes in the expressions of genes and proteins related to HMGB1-RAGE/TLRs-NF-κB signaling pathway in renal tissue. SV40 MES-13 mouse glomerular mesangial cells were induced to cell apoptosis by 50 mmol/L glucose, followed by intervention with HPS. Flow cytometry was used to measure cell apoptosis, and Western blotting was used to analyze changes in the expressions of proteins related to HMGB1-RAGE/TLRs-NF-κB signaling pathway. Results Compared with model group, the body weight and fasting blood glucose of mice in HPS group were significantly reduced (P < 0.001), and 24 h urine protein content was significantly reduced (P < 0.01, 0.001). The levels of insulin, total cholesterol, low-density lipoprotein cholesterol, creatinine and triglycerides in serum were significantly reduced (P < 0.05, 0.01, 0.001), while the level of high-density lipoprotein cholesterol was significantly increased (P < 0.001). The levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and IL-1β in serum were significantly reduced (P < 0.05, 0.01, 0.001). Renal pathological damage was significantly improved. The expression levels of genes and proteins related to HMGB1-RAGE/TLRs-NF-κB signaling pathway in renal tissue were significantly reduced (P < 0.05, 0.01, 0.001). The cell experiment results showed that HPS significantly inhibited high-glucose induced apoptosis of SV40 MES-13 cells (P < 0.05, 0.001), as well as the activation of HMGB1-RAGE/TLRs-NF-κB signaling pathway and the production of inflammatory factors (P < 0.05, 0.01, 0.001). Conclusion HPS could alleviate DKD by inhibiting the activation of HMGB1-RAGE/TLRs-NF-κB signaling pathway and reducing the expressions of inflammatory factors.

R285.5

咸阳市科技计划项目（L2003-ZDYF-SF-027）；咸阳职业技术学院科研基金项目（2023KJB01）