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山东大学学报 (医学版) ›› 2019, Vol. 57 ›› Issue (6): 51-60.doi: 10.6040/j.issn.1671-7554.0.2019.423

• 基础医学 • 上一篇    

扶肝化纤汤对肝纤维化大鼠TGF-β1/Smad信号通路的影响

肖政华1,杨辉2,雷伟3,杨君3,易旭4,谭敏敏5   

  1. 1. 贵州中医药大学第二临床医学院消化内科, 贵州 贵阳 550001;2.贵州中医药大学第二临床医学院神经内科, 贵州 贵阳 550001;3.贵州中医药大学2016级研究生, 贵州 贵阳 550001;4.贵州中医药大学第二临床医学院中心实验室, 贵州 贵阳 550001;5.贵州中医药大学第二临床医学院呼吸内科, 贵州 贵阳 550001
  • 发布日期:2022-09-27
  • 通讯作者: 肖政华. E-mail: 435204673@qq.com
  • 基金资助:
    国家自然科学基金(81660790);贵州省科学技术厅贵阳中医学院联合资金项目(黔科合LH字［2014］7318号);院内博士启动基金课题

Effect of Fugan Huaxian Decoction on TGF-β1/Smad signaling pathway in rats with hepatic fibrosis

XIAO Zhenghua1, YANG Hui2, LEI Wei3, YANG Jun3, YI Xu4, TAN Minmin5   

  1. 1. Department of Gastroenterology, Second Clinical Medical College of Guizhou University of Traditional Chinese Medicine, Guiyang 550001, Guizhou, China;
    2. Department of Neurology, the Second Clinical Medical College of Guizhou University of Traditional Chinese Medicine, Guiyang 550001, Guizhou, China;
    3. Grade 2016 Postgraduate, Guizhou University of Traditional Chinese Medicine, Guiyang 550001, Guizhou, China;
    4. Central laboratory of the Second Clinical Medical College of GuiZhou University of 〓山〓东〓大〓学〓学〓报〓(医〓学〓版)57卷6期〓-肖政华, 等.扶肝化纤汤对肝纤维化大鼠TGF-β1/Smad信号通路的影响〓\=-Traditional Chinese Medicine, Guiyang 550001, Guizhou, China;
    5. Department of Respiratory Medicine, the Second Clinical Medical College of GuiZhou University of Traditional Chinese Medicine, Guiyang 550001, Guizhou, China
  • Published:2022-09-27

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摘要： 目的 研究扶肝化纤汤对正虚毒蕴血瘀病证结合肝纤维化(HF)模型大鼠TGF-β1/Smad信号通路的影响,探讨其抗HF的作用机制。 方法 清洁级SD大鼠120只,随机等分为正常组、模型组、秋水仙碱组、扶肝化纤汤高、中、低剂量组。除正常组外,其余各组在经典四氯化碳(CCL4)皮下注射制备大鼠HF模型的基础上,以中医理论指导,建立正虚毒蕴血瘀型HF大鼠模型。造模6周验证模型,造模成功后,给药组相应灌胃给药,正常组和模型组给予等体积纯净水,均连续灌胃3周。末次灌胃的次日,检测各组大鼠血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、白蛋白(ALB)、羟脯氨酸(HyP)、层粘连蛋白(LN)、Ⅲ型前胶原肽(PⅢNP)的含量,肝组织行HE染色和Masson染色。用qRT-PCR、免疫组化法检测各组肝组织转化生长因子β1(TGF-β1)、果蝇抗生物皮肤生长因子蛋白3(Smad3)、果蝇抗生物皮肤生长因子蛋白7(Smad7)及其mRNA的表达。 结果 病理结果显示,与模型组比较,各给药组肝细胞损伤程度减轻,胶原纤维的增生减少;HE染色结果显示,肝纤维化程度分期好转(χ2=54.722,P<0.01);Masson染色结果显示,肝脏胶原纤维面积占比减少(χ2=52.149,P<0.01)。与正常组比较,模型组大鼠血清ALT(χ2=39.685,P<0.01)、AST(F=659.984,P<0.01)、Hyp(χ2=38.613,P<0.01)、LN(χ2=50.057,P<0.01)、PⅢNP(F=63.492,P<0.01)升高、ALB(χ2=51.908,P<0.01)下降,肝组织TGF-β1(χ2=40.900,P<0.01)、Smad 3(χ2=38.253,P<0.01)、TGF-β1 mRNA(F=92.304,P<0.01)、Smad 3 mRNA(F=119.563,P<0.01)表达升高,Smad 7(χ2=48.949,P<0.01)及Smad 7 mRNA(F=42.120,P<0.01)表达降低;与模型组比较,扶肝化纤汤不同剂量组均抑制上述指标的改变(P<0.01),且呈现出扶肝化纤汤高剂量组优于其余给药组(P<0.01)。 结论 正虚毒蕴血瘀病症结合HF模型大鼠建模成功。扶肝化纤汤具有保护肝细胞、减轻肝损伤、抑制HF的作用。扶肝化纤汤能够调控TGF-β1、Smad3、Smad7及其mRNA的表达,可能是其抗HF的作用机制之一。

关键词: 扶肝化纤汤, TGF-β1/Smad信号通路, 肝纤维化, 病证结合, 模型大鼠

Abstract: Objective To explorethe effect of Fugan Huaxian Decoction on transforming growth factor β1(TGFβ1)/Smad signaling pathway in rats with hepatic fibrosis(HF)and the mechanism of anti-hepatic fibrosis. Methods A total of 120 SD rats were randomly divided into control group, model group, colchicine group, and high, medium and low dose group of decoction. Except for the control group,the other groups established HF rat models of Qi deficiency, poison weakness and blood stasis under the guidance of traditional Chinese medicine(TCM)theory based on classical subcutaneous injection of carbon tetrachloride(CCl4). After 6 weeks, when the models were successfully established, the decoction groups received gastrogavage for 3 weeks, while the control group and model group were given equal volume of pure water for 3 weeks. Next day after the last gastrogavage, the serum aspartate aminotransferase(AST), alanine aminotransferase(ALT), albumin(ALB), hydroxyproline(Hyp),laminin(LN)and pre-collagen type Ⅲ(PⅢNP)levels were measured. The liver tissues were stained with HE and Mason stainings. The protein and mRNA expressions of TGF-beta1, Smad3 and Smad7 in liver tissues were detected with qRT-PCR and immunohistochemistry. Results Compared with the model group, decoction groups showed relieved hepatocyte injury, decreased collagen fiber proliferation, improvedhepatic fibrosis(χ2=54.722, P<0.01), and reduced proportion of liver collagen fibers(χ2=52.149, P<0.01). Compared with the control group, the model group had increased serum ALT(χ2=39.685, P<0.01), AST(F=659.984, P<0.01), Hyp(χ2=38.613, P<0.01), LN(χ2=50.057, P<0.01), PⅢNP(F=63.492, P<0.01), decreased ALB(χ2=51.908, P<0.01), increased expressions of TGF-β1(χ2=40.900, P<0.01), Smad3(χ2=38.253, P<0.01), TGF-β1 mRNA(F=92.304, P<0.01), Smad3 mRNA(F=119.563, P<0.01), and decreased Smad7(χ2=48.949, P<0.01)and Smad7 mRNA(F=42.120, P<0.01). Compared with the model group, the decoction groups inhibited the changes of the above indexes(P<0.01), especially the high dosage group(P<0.01). Conclusion The model of Qi deficiency, poison weakness and blood stasis complicated with HF was successfully established. Fugan Huaxian Decoction can protect hepatocytes, alleviate liver injury and inhibit HF, probably by regulatingthe expressions of TGF-beta1, Smad3 and Smad7.

Key words: Fugan Huaxian Decoction, TGF-β1/Smad signaling pathway, Hepatic fibrosis, Syndrome combination, Rat model

中图分类号: 

  • R574
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