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目的：基于Ang-Tie2通路探讨加减三甲散抗肝纤维化病理性血管新生的作用机制。方法：将小鼠随机分为正常对照组、模型对照组、加减三甲散1、2 g/kg组、阳性对照(甲苯磺酸索菲拉尼片0.12 g/kg)组，每组9只。除正常对照组，其余各组采用10%CCl4-橄榄油溶液(5 mL/kg)腹腔注射的方法建立肝纤维化模型。造模成功后给药各组灌胃相对应药物，正常对照组和模型对照组灌胃生理盐水。HE染色、Masson染色观察肝组织炎症及纤维化；检测血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、透明质酸(HA)、层粘连蛋白(LN)、Ⅲ型前胶原(PCⅢ)、Ⅳ型胶原蛋白(ColIV)的含量；Western blot法检测血管生成素1(Ang1)、血管生成素2(Ang2)、酪氨酸激酶受体(Tie2)、磷酸化酪氨酸激酶受体Tie2(p-Tie2)、磷酸化的蛋白激酶B(p-AKT)、内皮细胞型一氧化氮合酶(eNOS)、血管内皮生长因子A(VEGF-A)、血管性血友病因子(vWF)、分化抗原簇31(CD31)的蛋白表达；ELISA法检测肝组织Ang1、Ang2分泌量；qRT-PCR法检测Ang1、Ang2、Tie2的mRNA表达；免疫荧光染色法检测VEGF-A、vWF、CD31的荧光表达水平；免疫组化法检测Ang1、Ang2、Tie2的蛋白阳性表达水平。结果：与正常对照组比较，模型对照组血清AST、ALT活力、HA、LN、PCⅢ、ColIV含量明显升高(P＜0.01)，可见大量炎症细胞浸润及胶原纤维沉积；Ang1、Tie2 的mRNA表达明显降低(P＜0.05)，Ang2的mRNA表达明显上调(P＜0.01)，Ang1、Tie2、p-Tie2、p-AKT、eNOS的蛋白表达下调(P＜0.01)，Ang2、VEGFA、vWF、CD31的蛋白表达明显上调(P＜0.01)；与模型对照组比较，加减三甲散各组血清AST、ALT活力、HA、LN、PCⅢ、ColIV含量明显降低(P＜0.05)，胶原纤维沉积和炎性细胞浸润减少，Ang1、Tie2 的mRNA表达明显上调(P＜0.05)，Ang2 mRNA表达明显下调(P＜0.01)，Ang1、Tie2、p-Tie2、p-AKT、eNOS的蛋白表达显著上调(P＜0.01)，Ang2、VEGFA、vWF、CD31的蛋白表达明显下调(P＜0.05)。结论：加减三甲散可能通过调控Ang-Tie2通路抑制病理性血管新生，有效改善肝纤维化。
Abstract：Objective: To investigate the mechanism through which modified Sanjia Powder inhibits pathological angiogenesis in liver fibrosis based on the angiopoietin (Ang)-tyrosine kinase receptor 2 (Tie2) signaling pathway. Methods: Mice were randomized into a normal control group， a model control group， modified Sanjia Powder (2 g/kg and 1 g/kg)， and a positive control (Sorafenib Tosylate Tablets， 0.12 g/kg) group， with 9 mice in each group. Except the normal control group， the remaining groups were modeled for liver fibrosis via intraperitoneal injection of 10% CCl4-olive oil solution at a dose of 5 mL/kg. After successful modeling， each group received corresponding drugs by gavage， while the normal control and model control groups received normal saline. Hematoxylin-eosin (HE) staining and Masson staining were performed to observe hepatic inflammation and collagen deposition. Serum levels of alanine transaminase (ALT)， aspartate transaminase (AST)， hyaluronic acid (HA)， laminin (LN)， type III procollagen (PCⅢ)， and type IV collagen (ColIV) were quantified. Western blot was adopted to determine the protein levels of angiopoietin-1 (Ang1)， angiopoietin-2 (Ang2)， Tie2， phosphorylated Tie2 (p-Tie2)， phosphorylated protein kinase B (p-AKT)， endothelial nitric oxide synthase (eNOS)， vascular endothelial growth factor A (VEGF-A)， von Willebrand factor (vWF)， and cluster of differentiation 31 (CD31). Enzyme-linked immunosorbent assay (ELISA) was employed to measure the secretion levels of Ang1 and Ang2 in the liver tissue. qRT-PCR was employed to determine the mRNA levels of Angpt1， Angpt2， and Tek. Immunofluorescence staining was used to evaluate the fluorescence intensity of VEGF-A， vWF and CD31， and immunohistochemistry was performed to detect the positive expression of Ang1， Ang2， and Tie2. Results: Compared with the normal control group， the model control group exhibited elevated serum levels of AST， ALT， HA， LN， PCⅢ， and ColIV (P<0.01)， massive inflammatory cell infiltration and collagen fiber deposition， downregulated mRNA levels of Angpt1 and Tek (P<0.05)， upregulated mRNA level of Angpt2 (P<0.01)， downregulated protein levels of Ang1， Tie2， p-Tie2， p-AKT， and eNOS (P<0.01)， and upregulated protein levels of Ang2， VEGF-A， vWF， and CD31 (P<0.01). Compared with the model control group， each treatment group showed declined serum levels of AST， ALT， HA， LN， PCⅢ， and ColIV (P<0.05)， alleviated inflammatory infiltration and collagen deposition， upregulated mRNA levels of Angpt1 and Tek (P<0.05)， downregulated mRNA level of Angpt2 (P<0.01)， upregulated protein levels of Ang1， Tie2， p-Tie2， p-AKT， and eNOS (P<0.01)， and downregulated protein levels of Ang2， VEGF-A， vWF， and CD31 (P<0.05). Conclusion: Modified Sanjia Powder can inhibit pathological angiogenesis by regulating the Ang-Tie2 signaling pathway， thereby effectively ameliorating liver fibrosis.
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基本信息:
DOI：10.13412/j.cnki.zyyl.20260828.002
中图分类号:R285.5
引用信息:
[1]李国花,付慧玲,宋安东,等.基于Ang-Tie2通路探讨加减三甲散抗肝纤维化病理性血管新生的作用机制[J].中药药理与临床().DOI:10.13412/j.cnki.zyyl.20260828.002.
基金信息:
宁夏自然科学基金项目（编号：2023AAC05040）; 宁夏医科大学校级科研项目（编号：XT2022015）; 宁夏医科大学中医学院一流学科建设项目资助(编号：ZY0019110305); 国家中医药管理局高水平中医药重点学科建设项目资助(编号：zyyzdxk-2023209)
2026-08-31
2026-08-31
2026-08-31