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目的:本研究旨在探讨益肾健脾化瘀汤对糖尿病大鼠肝损伤的影响及机制。方法:通过高脂饲料喂养结合单次腹腔注射链脲佐菌素(STZ)35 mg/kg建立糖尿病大鼠模型，将造模成功的糖尿病大鼠随机分为模型对照组、益肾健脾化瘀汤16.8、33.6 g/kg组、阳性对照药卡格列净9 mg/kg组，药物连续干预8 w。以普通维持饲料喂养的正常大鼠为正常对照组。生化法检测血糖、血脂、肝功能、氧化应激、炎症因子相关指标；化学发光法检测肝纤四项；计算肝指数；HE染色、糖原染色、马松染色和透射电子显微镜观察肝脏病理变化；原位末端标记法(TUNEL)检测肝细胞凋亡；免疫组织化学染色法检测肝组织胶原沉积情况；蛋白质免疫印记法检测肝组织kelch样ECH关联蛋白1/核因子-E2相关因子2/血红素加氧酶(Keap1/Nrf2/HO-1)信号通路相关蛋白、凋亡相关蛋白和转化生长因子-β1(TGF-β1)/Smad信号通路相关蛋白的表达。结果:与正常对照组比较，模型对照组大鼠体质量显著降低，血糖、血脂和肝脏指数明显升高(P<0.05或P<0.01),大鼠血清中谷丙转氨酶(ALT)和谷草转氨酶(AST)活力明显升高(P<0.05或P<0.01),肝组织呈现脂肪变性及纤维化改变，血清中超氧化物歧化酶(SOD)活力、谷胱甘肽过氧化物酶(GSH-PX)和过氧化氢酶(CAT)含量显著降低(P<0.05或P<0.01),丙二醛(MDA)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、透明质酸酶(HA)、层粘连蛋白(LN)、IV型胶原(CIV)和III型前胶原(PCIII)含量明显升高(P<0.05或P<0.01),大鼠肝组织TUNEL染色阳性区域面积显著增加(P<0.01),肝组织Keap1蛋白表达明显上调(P<0.05),Nrf2和下游HO-1蛋白表达明显下调(P<0.05或P<0.01),半胱氨酸蛋白酶3(Caspase3)和B细胞淋巴瘤基因2(Bcl2)蛋白表达显著下调(P<0.05),Bcl2关联X蛋白质(BAX)的表达明显上调(P<0.05),肝组织纤维化相关因子α-平滑肌肌动蛋白(α-SMA)、转化生长因子-β1(TGF-β1)、纤连蛋白和1型胶原蛋白表达显著上调(P<0.01),TGF-β1/Smad信号通路相关蛋白表达显著上调；与模型对照组比较，益肾健脾化瘀汤各组大鼠的体质量升高，血糖、血脂和肝脏指数降低(P<0.05或P<0.01),血清中ALT、AST活力降低(P<0.05或P<0.01)、肝组织病理损伤减轻，血清中SOD活力、GSH-PX和CAT含量升高(P<0.01),MDA、TNF-α、IL-1β、HA、LN、CIV和PCIII含量降低，肝组织TUNEL染色阳性区域面积降低(P<0.05或P<0.01),肝组织Keap1蛋白表达下调(P<0.05),Nrf2、HO-1蛋白表达上调(P<0.01),Caspase3和BCL2蛋白表达上调(P<0.05),BAX蛋白表达下调(P<0.05),α-SMA、TGF-β1、纤连蛋白和1型胶原蛋白阳性表达水平降低(P<0.05或P<0.01),TGF-β1/Smad信号通路相关蛋白的表达下调(P<0.05或P<0.01)。结论:益肾健脾化瘀汤可以减轻糖尿病大鼠糖脂代谢紊乱和肝损伤，其机制可能与抑制氧化应激、细胞凋亡和纤维化信号通路有关。
Abstract：Objective:To investigate the effect of Yishen Jianpi Huayu(益肾健脾化瘀) Decoction on liver injury in diabetic rats and reveal the underlying mechanism. Methods:The rat model of diabetes was established by a high-fat diet combined with a single intraperitoneal injection of streptozotocin(STZ,35 mg/kg). The successfully modeled diabetic rats were randomly assigned into model control, Yishen Jianpi Huayu Decoction(16.8 g/kg and 33.6 g/kg),and positive control(canagliflozin, 9 mg/kg) groups and received drug intervention for 8 weeks. The rats being fed a standard maintenance diet were included as the normal control group. Biochemical methods were used to determine the levels of blood glucose, blood lipids, liver function indicators, oxidative stress indicators, inflammatory factors. The chemiluminescent assay was employed to measure the levels of four liver fibrosis indicators. The liver index was calculated. Hematoxylin-eosin staining, periodic acid-Schiff staining, Masson staining, and transmission electron microscopy were employed for pathological analysis of the liver tissue. TdT-mediated dUTP nick-end labeling(TUNEL) was employed to detect the apoptosis of rat hepatocytes. Collagen deposition in hepatocytes was detected by immunohistochemical staining. The expression levels of proteins in the Kelch-like ECH-associated protein 1/nuclear factor erythroid 2-related factor 2/heme oxygenase-1(Keap1/Nrf2/HO-1) signaling pathway, apoptosis-associated proteins, and proteins in the transforming growth factor-β1(TGF-β1)/Smad signaling pathway in the liver were analyzed by Western blotting. Results:Compared with the normal control group, the model control group showed decreased body weight, increased blood glucose level, blood lipid levels, and liver index(P<0.05 or P<0.01),elevated serum levels of alanine aminotransferase(ALT) and aspartate aminotransferase(AST)(P<0.05 or P<0.01),steatosis and fibrosis in the liver tissue, lowered serum levels of superoxide dismutase(SOD),glutathione peroxidase(GSH-PX),and catalase(CAT)(P<0.05 or P<0.01),raised serum level of malondialdehyde(MDA)(P<0.01),up-regulated protein level of Keap1(P<0.05),and down-regulated protein levels of Nrf2 and the downstream HO-1(P<0.05 or P<0.01). In addition, the model control group showed elevated serum levels of tumor necrosis factor(TNF)-α and interleukin(IL)-1β(P<0.01),raised serum levels of hyaluronidase(HA),laminin(LN),collagen type IV(CIV),and procollagen type III(PCIII)(P<0.05 or P<0.01),increased TUNEL-stained positive area in the liver(P<0.01),down-regulated protein levels of cysteinyl aspartate-specific proteinase 3(caspase3) and B-cell lymphoma-2(Bcl2)(P<0.05),and up-regulated protein level of Bcl2-associated X protein(BAX)(P<0.05). In addition, the liver tissue in the model control group presented elevated levels of hepatic fibrosis-related factors such as α-smooth muscle actin(α-SMA),TGF-β1,fibronectin, and collagen type I(Collagen-1)(P<0.01) and significantly up-regulated expression of proteins in the TGF-β1/Smad signaling pathway. Compared with the model control group, Yishen Jianpi Huayu Decoction increased the body weight of rats, decreased the blood glucose level, blood lipid levels, and liver index(P<0.05 or P<0.01),lowered the serum levels of ALT and AST(P<0.05 or P<0.01),alleviated the pathological damage in the liver tissue, raised the serum levels of SOD,GSH-PX,and CAT(P<0.01),inhibited the production of MDA(P<0.05 or P<0.01),Keap1 protein expression was inhibited(P<0.05),up-regulated the protein levels of Nrf2 and downstream HO-1(P<0.01). In addition, the decoction reduced the serum levels of TNF-α and IL-1β(P<0.05 or P<0.01),significantly decreased the serum levels of HA,LN,CIV,and PCIII and the TUNEL-stained positive area(P<0.05 or P<0.01),up-regulated the protein levels of caspase3 and Bcl2(P<0.05),down-regulated the protein level of BAX(P<0.05). Furthermore, the decoction down-regulated the expression levels of α-SMA,TGF-β1,fibronectin, and collagen-1(P<0.05 or P<0.01) and the proteins in the TGF-β1/Smad signaling pathway to inhibit liver fibrosis(P<0.05 or P<0.01). Conclusion:Yishen Jianpi Huayu Decoction can alleviate glucose and lipid metabolism disorders and liver injury in diabetic rats by inhibiting oxidative stress, apoptosis, and fibrosis.
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基本信息:
DOI：10.13412/j.cnki.zyyl.20250929.012
中图分类号:R285.5
引用信息:
[1]杨世瑜,高思齐,肖雨,等.益肾健脾化瘀汤对糖尿病大鼠肝脏氧化应激、凋亡和纤维化的影响及机制[J].中药药理与临床,2026,42(07):23-34.DOI:10.13412/j.cnki.zyyl.20250929.012.
基金信息:
河北省自然科学基金中医药联合基金重点项目(编号：H2023209038); 河北省高等学校科学技术研究项目(编号：ZD2022141)
2025-09-30
2025-09-30
2025-09-30