中文English
ISSN 1001-5256 (Print)
ISSN 2097-3497 (Online)
CN 22-1108/R

留言板

尊敬的读者、作者、审稿人, 关于本刊的投稿、审稿、编辑和出版的任何问题, 您可以本页添加留言。我们将尽快给您答复。谢谢您的支持!

姓名
邮箱
手机号码
标题
留言内容
验证码

扶正化瘀方对肝硬化小鼠模型肝细胞消亡与再生的影响

朱亭亭 齐婧姝 郭亚楠 刘洪亮 陶艳艳 赵志敏 李正鑫 刘成海

引用本文:
Citation:

扶正化瘀方对肝硬化小鼠模型肝细胞消亡与再生的影响

DOI: 10.12449/JCH240417
基金项目: 

国家自然科学基金面上项目 (82274305)

伦理学声明:本研究方案于2020年6月19日经由上海中医药大学实验动物伦理委员会审批,批号:2020-618-45-07,符合实验室动物管理与使用准则。
利益冲突声明:本研究不存在任何利益冲突。
作者贡献声明:朱亭亭、刘洪亮、李正鑫负责课题设计、资料分析、撰写论文;齐婧姝、郭亚楠参与收集数据、修改论文;陶艳艳、赵志敏负责指导撰写,修改论文;李正鑫、刘成海负责拟定写作思路、指导撰写文章并最后定稿。
详细信息
    通信作者:

    李正鑫, zhengxinli1990@hotmail.com (ORCID: 0000-0002-2497-6951)

    刘成海, chenghai.liu@outlook.com (ORCID: 0000-0002-1696-6008)

Effect of Fuzheng Huayu prescription on hepatocyte extinction and regeneration in a mouse model of liver cirrhosis

Research funding: 

General Project of National Natural Science Foundation of China (82274305)

More Information
  • 摘要:   目的  探讨扶正化瘀方对纤维化肝脏肝细胞消亡与再生的影响,及其药物促进肝细胞再生的作用机制。  方法  以CCl4腹腔注射6周诱导建立肝硬化小鼠模型。正常对照组9只,模型组10只,索拉非尼组10只,扶正化瘀方组10只。自造模第4周起,扶正化瘀方组、索拉非尼组分别给予4.8 g/kg、4 mg/kg小鼠体质量相应药物灌胃,连续3周;正常组和模型组给予等体积羧甲基纤维素钠。检测血清肝功能;METAVIR评分系统评价肝组织炎症及纤维化分期;天狼星红染色和肝组织羟脯氨酸含量评价胶原沉积量;免疫组化检测Ⅳ型胶原、CD31与CD32b、Ki67、CyclinD1、谷氨酰胺合成酶(GS)、Wnt2以及HGF蛋白的表达;Western Blot检测肝组织Wnt2、LRP6、β-catenin、p-β-catenin、CyclinD1表达。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。  结果  与模型组比较,扶正化瘀方组和索拉非尼组血清ALT、AST水平和肝组织羟脯氨酸含量均降低(P值均<0.01),METAVIR评分减低(P值均<0.05);Ⅳ型胶原、CD31表达减少(P值均<0.05),CD32b表达增加(P<0.01);肝组织实质病变消亡数量减少,Ki67、CyclinD1表达上升(P值均<0.01);Wnt2、LRP6、β-catenin、CyclinD1蛋白表达水平上调、p-β-catenin表达显著下调(P值均<0.05);肝组织CD32b与Wnt2共染阳性细胞显著增多。  结论  扶正化瘀方可通过抑制肝窦毛细血管化,改善肝窦内皮细胞Wnt2外分泌功能,激活肝细胞再生相关Wnt/β-catenin信号通路,最终逆转肝硬化。

     

  • 图  1  扶正化瘀方对CCl4肝硬化小鼠模型肝组织炎症、胶原沉积的影响(×100)

    Figure  1.  Effect of FZHY on liver inflammation, collagen deposition in CCl4 mice (×100)

    图  2  扶正化瘀方对肝组织羟脯氨酸含量的影响

    Figure  2.  Effect of FZHY on hydroxyproline content in liver tissue

    图  3  扶正化瘀方对CCl4肝硬化小鼠模型肝组织Ⅳ型胶原、CD31、CD32b表达的影响

    Figure  3.  Effect of FZHY on the expression of collagen type Ⅳ, CD31 and CD32b in CCl4 mice

    图  4  扶正化瘀方对CCl4肝硬化小鼠模型肝组织Ⅳ型胶原、CD31、CD32b的影响(×200)

    Figure  4.  Effect of FZHY on the collagen type Ⅳ, CD31 and CD32b in CCl4 mice (×200)

    图  5  扶正化瘀方对CCl4肝硬化小鼠模型肝组织PEL程度的影响(GS免疫组化染色)

    Figure  5.  Effect of FZHY on PEL in CCl4 mice (GS immunohistochemical staining)

    图  6  扶正化瘀方对肝组织PEL数量和对肝组织中央静脉与汇管区距离的影响

    Figure  6.  Effect of FZHY on PEL number and distance between central vein and portal area of liver tissue

    图  7  扶正化瘀方对肝组织Ki67和HGF表达的影响

    Figure  7.  Effect of FZHY on the expression of Ki67 and HGF in CCl4 mice

    图  8  扶正化瘀方对CCl4肝硬化小鼠模型肝组织Ki67和HGF的影响(×200)

    Figure  8.  Effect of FZHY on Ki67 and HGF in CCl4 mice (×200)

    图  9  扶正化瘀方对模型小鼠LSEC中CD32b与Wnt2表达的影响(×200)

    注: CD32b,黄色;Wnt2,红色;DAPI:蓝色。

    Figure  9.  Effect of FZHY on the expression of CD32b and Wnt2 in mice(×200)

    图  10  扶正化瘀方对CCl4肝硬化小鼠模型Wnt/β-catenin信号通路相关蛋白的影响(×200)

    Figure  10.  Effect of FZHY on Wnt/β-catenin signaling pathway-related protein in CCl4 mice(×200)

    图  11  扶正化瘀方对Wnt2和CyclinD1表达的影响

    Figure  11.  Effect of FZHY on the expression of Wnt2 and CyclinD1 in CCl4 mice

    图  12  扶正化瘀方对Wnt/β-catenin信号通路相关蛋白的影响

    Figure  12.  Effect of FZHY on Wnt/β-catenin signaling pathway-related protein in CCl4 mice

  • [1] XU XY, DING HG, LI WG, et al. Chinese guidelines on the management of liver cirrhosis[J]. J Clin Hepatol, 2019, 35( 11): 2408- 2425. DOI: 10.3969/j.issn.1001-5256.2019.11.006.

    徐小元, 丁惠国, 李文刚, 等. 肝硬化诊治指南[J]. 临床肝胆病杂志, 2019, 35( 11): 2408- 2425. DOI: 10.3969/j.issn.1001-5256.2019.11.006.
    [2] MAGDALENO F, TREBICKA J. Selective LOXL2 inhibition: potent antifibrotic effects in ongoing fibrosis and fibrosis regression[J]. Gut, 2017, 66( 9): 1540- 1541. DOI: 10.1136/gutjnl-2016-313621.
    [3] HARRISON SA, ABDELMALEK MF, CALDWELL S, et al. Simtuzumab is ineffective for patients with bridging fibrosis or compensated cirrhosis caused by nonalcoholic steatohepatitis[J]. Gastroenterology, 2018, 155( 4): 1140- 1153. DOI: 10.1053/j.gastro. 2018.07.006.
    [4] QI JS, HU XD, LIU CH. Reversal mechanism of hepatic fibrosis[J]. Chin J Hepatol, 2022, 30( 6): 577- 582. DOI: 10.3760/cma.j.cn501113-20220504-00239.

    齐婧姝, 胡旭东, 刘成海. 肝纤维化的逆转机制[J]. 中华肝脏病杂志, 2022, 30( 6): 577- 582. DOI: 10.3760/cma.j.cn501113-20220504-00239.
    [5] LIU CH, ZHAO ZM, LYU J, et al. Advances in the understanding and treatment of liver fibrosis in traditional Chinese medicine[J]. J Clin Hepatol, 2019, 35( 4): 728- 733. DOI: 10.3969/j.issn.1001-5256.2019.04.006.

    刘成海, 赵志敏, 吕靖. 中医对肝纤维化逆转的认识与治疗[J]. 临床肝胆病杂志, 2019, 35( 4): 728- 733. DOI: 10.3969/j.issn.1001-5256.2019.04.006.
    [6] CHENG Q, LI N, CHEN M, et al. Fuzheng Huayu inhibits carbon tetrachloride-induced liver fibrosis in mice through activating hepatic NK cells[J]. J Ethnopharmacol, 2013, 145( 1): 175- 181. DOI: 10.1016/j.jep.2012.10.047.
    [7] PAN Q, WANG YQ, LI GM, et al. Fuzheng Huayu recipe ameliorates liver fibrosis by restoring balance between epithelial-to-mesenchymal transition and mesenchymal-to-epithelial transition in hepatic stellate cells[J]. Biomed Res Int, 2015, 2015: 935903. DOI: 10.1155/2015/935903.
    [8] ZHANG M, LIU HL, HUANG K, et al. Fuzheng Huayu recipe prevented and treated CCl4-induced mice liver fibrosis through regulating polarization and chemotaxis of intrahepatic macrophages via CCL2 and CX3CL1[J]. Evid Based Complement Alternat Med, 2020, 2020: 8591892. DOI: 10.1155/2020/8591892.
    [9] LIU HL, LV J, ZHAO ZM, et al. Fuzhenghuayu Decoction ameliorates hepatic fibrosis by attenuating experimental sinusoidal capillarization and liver angiogenesis[J]. Sci Rep, 2019, 9( 1): 18719. DOI: 10.1038/s41598-019-54663-4.
    [10] YANG T, SHEN DP, WANG QL, et al. Investigation of the absorbed and metabolized components of Danshen from Fuzheng Huayu recipe and study on the anti-hepatic fibrosis effects of these components[J]. J Ethnopharmacol, 2013, 148( 2): 691- 700. DOI: 10.1016/j.jep.2013.05.031.
    [11] JAMALL IS, FINELLI VN, QUE HS. A simple method to determine nanogram levels of 4-hydroxyproline in biological tissues[J]. Anal Biochem, 1981, 12( 1): 70- 75. DOI: 10.1016/0003-2697(81)90261-x.
    [12] WANLESS IR, NAKASHIMA E, SHERMAN M. Regression of human cirrhosis. Morphologic features and the genesis of incomplete septal cirrhosis[J]. Arch Pathol Lab Med, 2000, 124( 11): 1599- 607. DOI: 10.5858/2000-124-1599-ROHC.
    [13] STUECK AE, WANLESS IR. Hepatocyte buds derived from progenitor cells repopulate regions of parenchymal extinction in human cirrhosis[J]. Hepatology, 2015, 61( 5): 1696- 707. DOI: 10.1002/hep.27706.
    [14] WANLESS IR. The role of vascular injury and congestion in the pathogenesis of cirrhosis: the congestive escalator and the parenchymal extinction sequence[J]. Curr Hepatology Rep, 2020, 19: 40- 53. DOI: 10.1007/s11901-020-00508-y.
    [15] HYTIROGLOU P, THEISE ND. Regression of human cirrhosis: an update, 18 years after the pioneering article by Wanless et al[J]. Virchows Arch, 2018, 473( 1): 15- 22. DOI: 10.1007/s00428-018-2340-2.
    [16] XU XJ, LU LG. Basic and clinical studies on reversal of liver fibrosis after hepatitis C virus eradication[J]. Chin Hepatol, 2021, 26( 8): 840- 842. DOI: 10.3969/j.issn.1008-1704.2021.08.005.

    徐贤军, 陆伦根. 丙型肝炎病毒根除后肝纤维化逆转基础和临床研究[J]. 肝脏, 2021, 26( 8): 840- 842. DOI: 10.3969/j.issn.1008-1704.2021.08.005.
    [17] XU Y, ZHANG YG, WANG X, et al. Long-term antiviral efficacy of entecavir and liver histology improvement in Chinese patients with hepatitis B virus-related cirrhosis[J]. World J Gastroenterol, 2015, 21( 25): 7869- 7876. DOI: 10.3748/wjg.v21.i25.7869.
    [18] MARCELLIN P, GANE E, BUTI M, et al. Regression of cirrhosis during treatment with tenofovir disoproxil fumarate for chronic hepatitis B: a 5-year open-label follow-up study[J]. Lancet, 2013, 381( 9865): 468- 475. DOI: 10.1016/S0140-6736(12)61425-1.
    [19] DING BS, NOLAN DJ, BUTLER JM, et al. Inductive angiocrine signals from sinusoidal endothelium are required for liver regeneration[J]. Nature, 2010, 468( 7321): 310- 315. DOI: 10.1038/nature09493.
    [20] ZHAO L, JIN Y, DONAHUE K, et al. Tissue repair in the mouse liver following acute carbon tetrachloride depends on injury-induced Wnt/β-catenin signaling[J]. Hepatology, 2019, 69( 6): 2623- 2635. DOI: 10.1002/hep.30563.
    [21] HU S, MONGA SP. Wnt/β-catenin signaling and liver regeneration: Circuit, biology, and opportunities[J]. Gene Expr, 2021, 20( 3): 189- 199. DOI: 10.3727/105221621X16111780348794.
    [22] RUSSELL JO, MONGA SP. Wnt/β-catenin signaling in liver development, homeostasis, and pathobiology[J]. Annu Rev Pathol, 2018, 13: 351- 378. DOI: 10.1146/annurev-pathol-020117-044010.
    [23] XU LM, LIU P, SHEN XZ, et al. Guidelines for the diagnosis and treatment of liver fibrosis in integrative medicine practice(2019)[J]. J Clin Hepatol, 2019, 35( 7): 1444- 1449. DOI: 10.3969/j.issn.1001-5256.2019.07.007.

    徐列明, 刘平, 沈锡中, 等. 肝纤维化中西医结合诊疗指南(2019年版)[J]. 临床肝胆病杂志, 2019, 35( 7): 1444- 1449. DOI: 10.3969/j.issn.1001-5256.2019.07.007.
    [24] CHENG J, Modern molecular biology of the extracellular matrix·Extracellular matrix and clinical medicine[M]. Beijing: Science Press, 2012: 696- 711.

    成军. 现代细胞外基质分子生物学·细胞外基质与临床医学[M]. 北京: 科学出版社, 2012: 696- 711.
  • 加载中
图(12)
计量
  • 文章访问数:  155
  • HTML全文浏览量:  73
  • PDF下载量:  24
  • 被引次数: 0
出版历程
  • 收稿日期:  2023-03-02
  • 录用日期:  2023-06-21
  • 出版日期:  2024-04-11
  • 分享
  • 用微信扫码二维码

    分享至好友和朋友圈

目录

    /

    返回文章
    返回