小柴胡汤对化学性肝纤维化小鼠的保护作用

doi:10.12092/j.issn.1009-2501.2020.05.001

摘要/Abstract

摘要： 目的:探讨小柴胡汤(XCHD)对四氯化碳(CCl4)诱发的肝纤维化(HF)小鼠的保护作用。方法:采用持续12周,2次/周,20%CCl4小鼠背部皮下注射的方法,复制HF小鼠模型。造模首日起,每天1次,持续12周,灌胃给予3.90、7.80、15.60 g/kg XCHD和0.1 g/kg水飞蓟宾。实验结束后,处置小鼠,检测血清中ALT、AST含量;HE、Masson、天狼猩红染色以及透射电镜观察肝脏病理组织学损伤情况,并进行HF分级评分;免疫荧光、RT-qPCR和免疫印迹技术检测肝组织中α-肌动蛋白(α-SMA)和Ι型胶原(Collagen Ι)水平。结果:XCHD(7.80,15.60 g/kg)组不仅可明显降低血清中ALT、AST水平(P<0.05),减轻肝脏病理学损伤程度,减少胶原纤维沉积,还可显著抑制α-SMA、Collagen Ι mRNA和蛋白表达水平(P<0.05)。结论:XCHD对化学性HF小鼠具有很好的保护作用。

关键词: 小柴胡汤, 肝纤维化, 四氯化碳

Abstract: AIM: To observe the protective effects of Xiaochaihu Decoction (XCHD) on carbon tetrachloride (CCl4)-induced hepatic fibrosis (HF) in mice. METHODS: HF was induced in male mice by 20% CCl4 twice a week for 12 weeks. XCHD (3.90, 7.80, 15.60 g/kg) and Silybin (0.1 g/kg) were administrated via gavage once a day starting from the CCl4 treatment for subsequent 12 weeks. Then, the levels ALT and AST in serum were assayed, liver samples were taken to examine the degree of HF by HE, Masson, Sirius Red staining and Electron microscope. Moreover, the protein and mRNA expression levels of α-smooth muscle actin (α-SMA) and Collagen Ι were determined by RT-qPCR, immunofluorescence and Western blot. RESULTS: The data demonstrated that XCHD (7.80, 15.60 g/kg) effectively reduced histopathological changes, such as steatosis, collagen deposition; meanwhile the histopathological analysis suggested that XCHD obviously alleviated the degree of HF. Furthermore, XCHD significantly reduced the levels of ALT, AST (P<0.05), and attenuated the expressions of α-SMA, Collagen Ι both of protein and mRNA levels (P<0.05). CONCLUSION: XCHD has protective effect on chemical HF in mice.

Key words: Xiaochaihu Decoction, hepatic fibrosis, carbon tetrachloride, mice

中图分类号:

R965.2

吴芙蓉, 宁丽娟, 周冉. 小柴胡汤对化学性肝纤维化小鼠的保护作用[J]. 中国临床药理学与治疗学, 2020, 25(5): 481-488.

WU Furong, NING Lijuan, ZHOU Ran. Protective effects of Xiaochaihu Decoction on chemical hepatic fibrosis in mice[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2020, 25(5): 481-488.

参考文献

[1]Chen G, Xia B, Fu Q, et al. Matrix mechanics as regulatory factors and therapeutic targets in hepatic fibrosis [J]. Int J Biol Sci, 2019, 15(12): 2509-2521.
[2]Zhu J, Luo Z, Pan Y, et al. H19/miR-148a/USP4 axis facilitates liver fibrosis by enhancing TGF- beta signaling in both hepatic stellate cells and hepatocytes [J]. J Cell Physiol, 2019, 234(6): 9698- 9710.
[3]Martin-Mateos R, De Assuncao TM, Arab JP, et al. Enhancer of zeste homologue 2 inhibition attenuates TGF-beta dependent hepatic stellate cell activation and liver fibrosis [J]. Cell Mol Gastroenterol Hepatol, 2019, 7(1): 197-209.
[4]黄培龙, 叶彬华, 林莉. 基于"少阳主枢"谈小柴胡汤治疗代谢综合征的应用 [J].江西中医药大学学报, 2019, 31(2): 6-8.
[5]安兴, 翁祥文, 彭波. 从《伤寒论》谈小柴胡汤的临床运用 [J]. 河北中医, 2018, 40(10): 1578-1581.
[6]陈丹丹, 龚作炯. 小柴胡汤治疗慢性乙型肝炎肝纤维化的临床研究 [J]. 现代中西医结合杂志, 2011, 20(31): 3928-3930.
[7]李晋, 徐尚福, 李远洋, 等. 小柴胡汤对肝纤维化大鼠肝脏MMP-2、TIMP-2表达的影响 [J]. 辽宁中医杂志, 2018, 45(31): 1066-1068, 1119.
[8]陈靖, 谢鑫, 吴成举, 等. 经方小柴胡汤对小鼠肝纤维化的改善作用及其机制的研究 [J]. 中华中医药学刊, 2017, 35(5): 1271-1274, 1365.
[9]蔡皓, 庄延双, 刘晓, 等. 正交优选小柴胡汤的提取工艺 [J]. 中药材, 2012, 35(12): 2036-2039.
[10]许建明, 徐叔云, 张运芳, 等. 四氯化碳诱导小鼠肝纤维化模型的建立 [J].中国药理学通报, 2000, 16(3): 339-341.
[11]郑旭锐, 孙守才, 李长秦, 等. 姜黄素对肝纤维化模型肝组织Ⅰ、Ⅲ型胶原mRNA影响的研究 [J]. 中药材, 2007, 30(9): 1118-1120.
[12]Jin Z, Liu S, Zhan Q, et al. Decoy receptor 3 alleviates hepatic fibrosis through suppressing inflammation activated by NF-kappaB signaling pathway [J]. Adv Clin Exp Med, 2018, 27(4): 441-447.
[13]Horowitz JM, Venkatesh SK, Ehman RL, et al. Evaluation of hepatic fibrosis: a review from the society of abdominal radiology disease focus panel [J]. Abdom Radiol (NY), 2017, 42(8): 2037-2053.
[14]郭悦承, 陆伦根. 单核细胞趋化蛋白-1及其受体在肝纤维化进程中的作用 [J]. 临床肝胆病杂志, 2019, 35(12): 2793-2795.
[15]李妍, 陆伦根. 肝纤维化的分子基础与临床诊治现状 [J]. 临床肝胆病杂志, 2018, 34(1): 12-15.
[16]Zhu J, Zhang Z, Zhang Y, et al. MicroRNA-212 activates hepatic stellate cells and promotes liver fibrosis via targeting SMAD7 [J]. Biochem Bioph Res Co, 2018, 496(1): 176-183.
[17]Tian H, Liu L, Li Z, et al. Chinese medicine CGA formula ameliorates liver fibrosis induced by carbon tetrachloride involving inhibition of hepatic apoptosis in rats [J]. J Ethnopharmacol, 2019, 232: 227-235.
[18]郑琼娜, 武俏俏, 马哲乐, 等. 低分子肝素钠对小鼠实验性肝纤维化干预作用的初步研究 [J]. 中国临床药理学与治疗学, 2019, 24(5): 511-516.
[19]刘梦堃, 吴荣秀. 肝纤维化诊断新进展 [J]. 医学综述, 2014, 20(15): 2782-2783.
[20]Wang CW, Liao KW, Chan CC, et al. Association between urinary thiodiglycolic acid level and hepatic function or fibrosis index in school-aged children living near a petrochemical complex [J]. Environ Pollut, 2019, 244: 648-656.
[21]Lachowski D, Cortes E, Rice A, et al. Matrix stiffness modulates the activity of MMP-9 and TIMP-1 in hepatic stellate cells to perpetuate fibrosis [J]. Sci Rep, 2019, 9(1): 7299.
[22]Sferra R, Vetuschi A, Pompili S, et al. Expression of pro-fibrotic and anti-fibrotic molecules in dimethylnitrosamine-induced hepatic fibrosis [J]. Pathol Res Pract, 2017, 213(1): 58-65.
[23]Chen Q, Chen L, Kong D, et al. Dihydroartemisinin alleviates bile duct ligation-induced liver fibrosis and hepatic stellate cell activation by interfering with the PDGF-betaR/ERK signaling pathway [J]. Int Immunopharmacol, 2016, 34: 250-258.
[24]Cheng JH, She H, Han YP, et al. Wnt antagonism inhibits hepatic stellate cell activation and liver fibrosis [J]. Am J Physiol Gastrointest Liver Physiol, 2008, 294(1): G39-G49.