Li Yachun, Guo Shuai, Yang Fan, Liu Lifei, Chen Zhiqiang
Hebei University of Chinese Medicine, Shijiazhuang 050091, China.
Hebei Hospital of Traditional Chinese Medicine, Shijiazhuang 050091, China.
Evid Based Complement Alternat Med. 2021 Jul 28;2021:5873007. doi: 10.1155/2021/5873007. eCollection 2021.
Diabetic nephropathy (DN), a severe microvascular complication of diabetes, is one of the leading causes of end-stage renal disease. Huayu Tongluo Recipe (HTR) has been widely used in the clinical treatment of DN in China, and its efficacy is reliable. This study aimed to explore the renoprotective effect of HTR and the underlying mechanism. Male Sprague-Dawley rats were fed with high sugar and fat diet combined with an intraperitoneal injection of STZ to establish the diabetic model. Rats in each group were respectively given drinking water, HTR, and irbesartan by gavage for 16 weeks. 24-hour urine samples were collected every 4 weeks to detect the content of total protein and 8-OHdG. Blood samples were taken to detect biochemical indicators and inflammatory markers at the end of 16th week. Renal tissue was collected to investigate pathological changes and to detect oxidative stress and inflammatory markers. AMPK/Nrf2 signaling pathway and fibrosis-related proteins were detected by immunohistochemistry, immunofluorescence, real-time PCR, and western blot. 24h urine total protein (24h UTP), serum creatinine (Scr), blood urea nitrogen (BUN), total cholesterol (TC), and triglyceride (TG) were decreased in the rats treated with HTR, while there was no noticeable change of blood glucose. HTR administration decreased malondialdehyde (MDA) content and increased superoxide dismutase (SOD) activity in kidneys, complying with reduced 8-OHdG in the urine. The levels of TNF-, IL-1, and MCP1 and the expression of nuclear NFB were also lower after HTR treatment. Furthermore, HTR alleviated pathological renal injury and reduced the accumulation of extracellular matrix (ECM). Besides, HTR enhanced the AMPK/Nrf2 signaling and increased the expression of HO-1 while it inhibited the Nox4/TGF-1 signaling in the kidneys of STZ-induced diabetic rats. HTR can inhibit renal oxidative stress and inflammation to reduce ECM accumulation and protect the kidney through activating the AMPK/Nrf2 signaling pathway in DN.
糖尿病肾病（DN）是糖尿病严重的微血管并发症之一，是终末期肾病的主要病因之一。化瘀通络方（HTR）在中国已广泛用于DN的临床治疗，其疗效可靠。本研究旨在探讨HTR的肾脏保护作用及其潜在机制。雄性Sprague-Dawley大鼠给予高糖高脂饮食并腹腔注射链脲佐菌素以建立糖尿病模型。每组大鼠分别通过灌胃给予饮用水、HTR和厄贝沙坦，持续16周。每4周收集24小时尿液样本，检测总蛋白和8-羟基脱氧鸟苷的含量。在第16周结束时采集血样，检测生化指标和炎症标志物。收集肾组织，研究病理变化并检测氧化应激和炎症标志物。通过免疫组织化学、免疫荧光、实时PCR和蛋白质印迹法检测AMPK/Nrf2信号通路和纤维化相关蛋白。HTR治疗的大鼠24小时尿总蛋白（24h UTP）、血清肌酐（Scr）、血尿素氮（BUN）、总胆固醇（TC）和甘油三酯（TG）降低，而血糖无明显变化。给予HTR可降低肾脏中丙二醛（MDA）含量并增加超氧化物歧化酶（SOD）活性，与尿中8-OHdG减少一致。HTR治疗后肿瘤坏死因子-α、白细胞介素-1和单核细胞趋化蛋白-1水平以及核因子-κB的表达也较低。此外，HTR减轻了肾脏病理损伤并减少了细胞外基质（ECM）的积累。此外，HTR增强了AMPK/Nrf2信号传导并增加了血红素加氧酶-1（HO-1）的表达，同时抑制了链脲佐菌素诱导的糖尿病大鼠肾脏中的Nox4/转化生长因子-β1（TGF-β1）信号传导。在DN中，HTR可通过激活AMPK/Nrf2信号通路抑制肾脏氧化应激和炎症，以减少ECM积累并保护肾脏。
