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目的：基于AMPK-SIRT3-BNIP3信号通路探讨益气养阴活血方对糖尿病肾病（diabetic kidney disease,DKD）大鼠肾脏的保护机制。方法：采用SPF级雄性SD大鼠42只，体质量（200±20） g，经1周环境适应后，将其分为空白组（n=6）和造模组（n=36）,2组分别喂养普通饲料和高脂高糖饲料。造模组单次腹腔注射链脲佐菌素（35 mg/kg）建立DKD模型，将造模成功的大鼠分为缬沙坦（代文）组、益气养阴活血方等剂量组、益气养阴活血方高剂量组及模型组。各治疗组大鼠按照相应药物剂量灌胃，空白组与模型组予以等体积的蒸馏水，持续治疗7周。生化检测大鼠24 h尿蛋白定量（24 h-UTP）及血清生化指标：肾功能指标（SCr、BUN）、肝功能指标（ALB、ALT）、糖脂代谢指标（GLU、TC、TG）;H&E染色观察肾脏病理；Western blot检测肾组织AMPK、SIRT3、BNIP3蛋白表达水平；RT-PCR检测肾组织AMPK、SIRT3、BNIP3 mRNA表达水平。结果：与空白组比较，模型组大鼠ALT、Scr、BUN、TC、TG、GLU、24 h-UTP水平，肾组织BNIP3蛋白及mRNA表达水平明显增加（P<0.05）,ALB水平及肾组织SIRT3、AMPK蛋白及mRNA表达水平明显降低（P<0.05），肾脏病理损伤严重。与模型组比较，各治疗组大鼠ALT、Scr、BUN、TC、TG、GLU、24 h-UTP水平，肾组织BNIP3蛋白及mRNA水平表达明显下降（P<0.05）,ALB水平及肾组织AMPK、SIRT3蛋白及mRNA表达水平明显上升（P<0.05），肾脏病理损伤明显减轻。各治疗组间比较差异无统计学意义（P>0.05）。结论：益气养阴活血方可有效降低DKD大鼠24 h-UTP、保护肾功能、改善肾脏病理，从而延缓DKD的发生、发展，其机制可能与调控肾组织AMPK-SIRT3-BNIP3通路有关。
Abstract：Objective: To explore the protective mechanism of Yiqi Yangyin Huoxue Recipe on diabetic kidney disease(DKD) rats based on AMPK-SIRT3-BNIP3 signaling pathway. Methods: A total of 42 SPF-grade male SD rats(with body weight ranging from 200～220 g) were used. After 1 week of environmental adaptation, the rats were divided into a blank group(n=6) and a modeling group(n=36). The two groups were fed with normal diet and high-fat/high-sugar diet respectively. A single intraperitoneal injection of streptozotocin(35 mg/kg) was used to establish the DKD model in the modeling group. The rats that were successfully induced were randomly divided into a valsartan(Diovan) group, a Yiqi Yangyin Huoxue Recipe equal-dose group, a Yiqi Yangyin Huoxue Recipe high-dose group, and a model group. The rats in each treatment group were given the corresponding drug doses by gavage, while the blank group and the model group were given the same volume of distilled water. The treatment lasted for 7 weeks. The 24-hour urinary protein(24 h-UTP) and serum biochemical indexes of the rats were detected by biochemical tests, including renal function indexes(SCr and BUN), liver function indexes(ALB and ALT),and glycolipid metabolism indexes(GLU, TC, and TG); H&E staining was used to observe the renal pathology; Western blot was used to detect the protein expression levels of AMPK, SIRT3, and BNIP3 in renal tissue; RT-PCR was used to detect the mRNA expression levels of AMPK, SIRT3, and BNIP3 in renal tissue. Results: Compared to the blank group, the levels of ALT, Scr, BUN, TC, TG, GLU, 24 h-UTP, and renal tissue BNIP3 protein and mRNA expression levels in the model group rats were significantly increased(P<0.05), while the levels of ALB and renal tissue SIRT3 and AMPK proteins and mRNA expression levels were significantly decreased(P<0.05), and the renal pathological damage was severe. Compared to the model group, the levels of ALT, Scr, BUN, TC, TG, GLU, 24 h-UTP, and renal tissue BNIP3 protein and mRNA expression levels in the treatment groups significantly decreased(P<0.05), while ALB level and renal tissue AMPK and SIRT3 protein and mRNA expression levels significantly increased(P<0.05), and the renal pathological damage was significantly alleviated. There was no statistically significant difference among the treatment groups(P>0.05). Conclusion: The Yiqi Yangyin Huoxue Recipe can effectively reduce 24 h-UTP in DKD rats, protect renal function, and improve renal pathology, thereby delaying the occurrence and development of DKD. The mechanism may be related to the regulation of the AMPK-SIRT3-BNIP3 pathway in renal tissue.
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基本信息:
DOI：10.13210/j.cnki.jhmu.20250514.002
中图分类号:R285.5
引用信息:
[1]李慧聪,邢静静,宋珂,等.基于AMPK-SIRT3-BNIP3通路探讨益气养阴活血方对糖尿病肾病大鼠作用机制[J].海南医科大学学报,2026,32(08):595-601.DOI:10.13210/j.cnki.jhmu.20250514.002.
基金信息:
国家自然科学基金（82074493）; 河南省卫健委国家中医临床研究基地科研专项（2022JDZX003）; 河南省中医药科学研究重大专项（2023ZYZD02）; 河南省中医药学科领军人才项目[豫卫中医函（2021）8号]; 河南省“双一流”创建工程（HSRP-DFCTCM-2023-3-07）~~
2025-05-14
2025-05-14
2025-05-14