基于TNF-α/NF-κB信号通路探讨益气活血方对糖尿病肾脏病大鼠肾脏的保护作用

Exploring the protective effect of Yiqi Huoxue Recipe on the kidneys of diabetic kidney disease rats based on the TNF-α/NF-κB signaling pathway

关键词:  糖尿病肾脏病  NF-κB信号通路  益气活血方  炎症

Key words:  diabetic kidney disease  NF-κB signaling pathway  Yiqi Huoxue Recipe  inflammation

摘要:

[目的] 观察益气活血方干预糖尿病肾脏病（DKD）大鼠后肿瘤坏死因子-α（TNF-α）/核转录因子-κB（NF-κB）信号通路的表达变化探讨益气活血方对DKD大鼠肾脏的保护作用。[方法] 雄性SD大鼠通过腹腔注射链脲菌素（STZ）并联合高糖高脂饲养建立DKD模型，剔除死亡及未成模大鼠，设置模型组、厄贝沙坦组、益气活血方高、中、低剂量组及对照组。各组大鼠按对应药物/纯水连续灌胃干预8周。干预阶段，持续观察记录一般状态、随机血糖（RBG）、体质量、日摄食量、日饮水量及24 h尿微量白蛋白（MAU）含量。干预结束后，检测血清中的尿素氮（BUN）、空腹血糖（FBG）、血肌酐（SCr）、总胆固醇（TC）及三酰甘油（TG）含量，并计算肾脏肥大指数（RHI）；采用苏木精-伊红（HE）染色观察肾脏病理变化，免疫组化和蛋白免疫印（Western blot）法检测NF-κB p65、TNF-α及细胞间黏附分子-1（ICAM-1）蛋白在肾组织的表达。[结果] 与对照组相比，模型组RBG显著升高（P<0.001）并出现体质量减少、多饮、多食及多尿等典型症状；其次，FBG、TG、TC、SCr、BUN、24 h MAU及RHI明显升高（P<0.01或P<0.001）；肾脏病理观察显示，肾小球肥大、肾小囊腔狭窄或固缩坏死，肾小管上皮细胞肿胀，部分呈空泡样变性，管腔显著变窄，并伴有大量炎性细胞浸润肾间质；肾脏组织中NF-κB p65、TNF-α及ICAM-1的蛋白表达均明显上调（P<0.001）。与模型组相比，益气活血方高剂量组显著降低DKD大鼠的RBG、FBG、24 h MAU、BUN及RHI（P<0.05或P<0.01）；减轻肾脏病理损伤，并抑制肾脏组织中NF-κB p65、TNF-α及ICAM-1的蛋白表达（P<0.05或P<0.001）。[结论] 益气活血方通过抑制TNF-α/NF-κB信号通路的表达，减轻肾脏的炎症反应，有效缓解肾脏病理损伤，发挥显著的肾保护作用。

Abstract:

[Objective] To observe changes in the expression of the TNF-α/NF-κB signaling pathway following intervention with the Yiqi Huoxue Recipe in diabetic kidney disease（DKD） rats and to explore its protective effects on the kidneys of DKD rats. [Methods] Male SD rats were intraperitoneally injected with streptozotocin（STZ） and fed a high-sugar，high-fat diet to establish a DKD model. Rats that died or failed to develop the model were excluded. The rats were divided into the control group，model group，irbesartan group，and High-dose，Medium-dose，and Low-dose Yiqi Huoxue Recipe groups of Yiqi Huoxue Recipe. Each group received continuous gavage treatment for 8 weeks with the corresponding drug or pure water.During the treatment phase，general health status，body weight，daily food intake，daily water intake，and random blood glucose（RBG） levels were continuously monitored. Urine samples were collected to measure 24-hour urinary microalbumin（MAU） levels. At the end of the treatment，fasting blood glucose（FBG），serum creatinine（SCr），urea nitrogen（BUN），triglycerides（TG），and total cholesterol（TC） levels were measured，and renal hypertrophy index（RHI） was calculated.Hematoxylin-eosin（HE） staining was performed to observe renal pathology，and immunohistochemistry and Western blotting were used to detect the expression of NF-κB p65，TNF-α，and ICAM-1 proteins in renal tissues. [Results] Compared to the control group，the model group exhibited significantly higher RBG（P<0.001） and typical symptoms including weight loss，polydipsia，polyphagia，and polyuria. Additionally，FBG，TG，TC，SCr，BUN，24 h MAU，and RHI were significantly elevated（P<0.01 or P<0.001）. Renal pathology revealed glomerular hypertrophy，narrowing or consolidation and necrosis of the renal capsule lumen，swelling of renal tubular epithelial cells，vacuolar degeneration，significant narrowing of the tubular lumen，and extensive inflammatory cell infiltration into the renal interstitium. The expression of NF-κB p65，TNF-α，and ICAM-1 was significantly upregulated in renal tissues（P<0.001）. Compared to the model group，the high-dose Yiqi Huoxue Recipe group significantly reduced RBG，FBG，24 h MAU，BUN，and RHI in DKD rats（P<0.05 or P<0.01），alleviated renal pathological damage，and inhibited the expression of NF-κB p65，TNF-α，and ICAM-1 proteins in renal tissues（P<0.05 or P<0.001）. [Conclusion] The Yiqi Huoxue Recipe inhibited the TNF-α/NF-κB signaling pathway，alleviating kidney inflammation and effectively reducing renal pathological damage，thereby exerting significant nephroprotective effects.