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

解毒化瘀通腑方对乙型肝炎肝硬化肝胆湿热证患者肠道菌群的影响

刘江凯 牛亚蒙 李素领 马庆亮 张建文 张雅儒 李冰倩

引用本文:
Citation:

解毒化瘀通腑方对乙型肝炎肝硬化肝胆湿热证患者肠道菌群的影响

DOI: 10.3969/j.issn.1001-5256.2022.04.016
基金项目: 

国家自然科学基金联合基金 (U1504825)

河南省高等学校重点科研项目计划 (20A360014)

河南省中医药拔尖人才培养项目 (Yuwei Zhongyi Han [2021] No.15)

伦理学声明:本研究方案于2019年12月9日经河南中医药大学第一附属医院伦理委员会审批,批号:2019HL-171,所有纳入患者均签署知情同意书。
利益冲突声明:本研究不存在研究者、伦理委员会成员、受试者监护人以及与公开研究成果有关的利益冲突。
作者贡献声明:刘江凯对研究的思路或设计有关键贡献;李素领、牛亚蒙参与病例资料收集;刘江凯、牛亚蒙、马庆亮参与研究数据的获取分析及解释;张建文、张雅儒、李冰倩参与数据汇总及指标检测。
详细信息
    通信作者:

    刘江凯, xmlc001@126.com

Effect of Jiedu Huayu Tongfu prescription on intestinal flora in patients with hepatitis B cirrhosis with liver-gallbladder damp-heat syndrome

Research funding: 

Joint Fund of National Natural Science Foundation of China (U1504825);

Key Scientific Research Project Plan of Colleges and Universities in Henan Province (20A360014);

Henan Provincial Top Talent Training Program of Traditional Chinese Medicine (Yuwei Zhongyi Han [2021] No.15)

More Information
    Corresponding author: LIU Jiangkai, xmlc001@126.com (ORCID: 0000-0002-1529-5089)
  • 摘要:   目的  研究解毒化瘀通腑方对乙型肝炎肝硬化肝胆湿热证患者肠道稳态的调节作用及对内毒素、炎性因子和细胞免疫功能的影响。  方法  纳入2019年6月至2021年1月在河南中医药大学第一附属医院就诊,符合诊断及纳入标准的72例患者为研究对象,随机分为观察组和对照组,每组各36例。其治疗组脱落2例,剔除2例,完成32例;对照组脱落2例,剔除1例,完成33例。两组均给予抗病毒、保肝等基础治疗,观察组加服解毒化瘀通腑颗粒,对照组给予双歧杆菌四联活菌片口服,疗程为4周。运用16S rDNA测序技术对两组粪便菌群进行基因测序,同时检测治疗前后肝功能(ALT、AST、TBil、Alb)、内毒素(ET)、TNFα、IL-6水平以及T淋巴细胞亚群(CD3+T、CD4+T、CD8+T、CD4+/CD8+)的变化。计量资料服从正态分布及方差齐性时,组内比较采用配对t检验,2组间比较用独立样本t检验,不服从正态分布时用Wilcoxon秩和检验;计数资料组间比较采用χ2检验。  结果  观察组临床总有效率为87.5%,对照组总有效率为60.6%,观察组总有效率明显高于对照组(χ2=-2.299,P=0.022)。两组治疗后ALT、AST、TBil均降低,Alb水平升高(P值均<0.05),与对照组相比,观察组TBil水平降低更显著(Z=-2.165,P=0.030)。两组患者治疗后CD3+T、CD4+T、CD4+/CD8+水平均明显改善, 且观察组较对照组更显著(Z值分别为-2.146、-2.940、3.157,P值分别为0.032、0.003、0.002)。两组患者治疗后炎性因子TNFα、IL-6与ET水平均明显下降,且观察组优于对照组(Z值分别为-2.139、-1.982、-2.062,P值分别为0.032、0.048、0.043)。两组治疗后OTU数量均较前上升。肠道菌群结构丰度在门水平上,观察组治疗后厚壁菌门丰度的上升和拟杆菌门丰度的下降较治疗前更显著(Z值分别为-3.181、-2.215,P值分别为0.001、0.027);与对照组相比,观察组厚壁菌门、蓝细菌门丰度的升高和拟杆菌门、梭菌门、ε-变形菌门的降低更明显(P值均<0.05);在属水平上,观察组双歧杆菌属较治疗前升高更明显(Z=-2.045,P=0.041)。Alpha多样性分析表明,观察组Chao1、Ace指数较治疗前升高更加显著(t值分别为-4.263、-3.328,P值分别为0.001、0.005),其中Ace指数较对照组上升更明显(t=2.292,P=0.030)。Beta多样性分析表明,各组菌群构成相似,无明显统计学差异(P值均>0.05)。  结论  在病因及基础治疗的基础上联合解毒化瘀通腑方治疗能够缓解乙型肝炎肝硬化肝胆湿热证患者临床症状,减轻肝损伤,改善细胞免疫功能。解毒化瘀通腑方可能通过增加厚壁菌门、乳杆菌属、双歧杆菌属等有益菌的丰度和减少拟杆菌门、梭菌门等致病菌的丰度来改善患者肠道菌群失调,其对肝脏和免疫功能的进一步改善作用可能与调整肠道微生态有关。

     

  • 图  1  组间Venn图

    注: ZYQ为观察组治疗前; ZYH为观察组治疗后; XYQ为对照组治疗前; XYH为对照组治疗后。

    Figure  1.  Venn diagram between groups

    图  2  肠道菌群结构丰度柱状图

    注: a, 门水平上的差异; b, 属水平上的差异。ZYQ为观察组治疗前, ZYH为观察组治疗后, XYQ为对照组治疗前, XYH为对照组治疗后。

    Figure  2.  Structural abundance histogram of intestinal flora

    图  3  Alpha多样性分析图

    注: ZYQ为观察组治疗前; ZYH为观察组治疗后; XYQ为对照组治疗前; XYH为对照组治疗后。

    Figure  3.  Alpha diversity analysis

    图  4  肠道菌群结构PCoA、NMDS分析图

    注: a, PCoA分析图; b, NMDS分析图。ZYQ为观察组治疗前, ZYH为观察组治疗后, XYQ为对照组治疗前, XYH为对照组治疗后。

    Figure  4.  Analysis of intestinal flora structure PCoA and NMDS

    表  1  两组中医症状疗效比较

    Table  1.   Comparison of curative effects of TCM symptoms between the two groups

    组别 例数 显效[例(%)] 有效[例(%)] 无效[例(%)] 总有效率(%)
    观察组 32 7(21.9) 21(65.6) 4(12.5) 87.5
    对照组 33 4(12.1) 16(48.5) 13(39.4) 60.6
    下载: 导出CSV

    表  2  两组肝功能比较

    Table  2.   Comparison of liver function between the two groups

    组别 例数 ALT(U/L) AST(U/L)
    治疗前 治疗后 Z P 治疗前 治疗后 Z P
    观察组 32 44.45(36.78~66.38) 32.50(27.45~44.08) 7.007 < 0.001 48.85(33.15~63.33) 37.40(29.93~42.53) -4.507 < 0.001
    对照组 33 47.50(33.75~65.30) 33.80(24.80~49.55) 5.599 < 0.001 46.20(33.40~65.00) 35.10(26.85~50.25) -4.083 < 0.001
    Z -0.046 -0.387 -0.092 -0.013
    P 0.963 0.699 0.927 0.990
    组别 例数 TBil(μmol/L) Alb(g/L)
    治疗前 治疗后 Z P 治疗前 治疗后 Z P
    观察组 32 20.08(17.25~39.75) 15.80(12.85~21.23) -4.506 < 0.001 35.50(30.35~41.83) 38.25(34.60~43.78) -2.940 0.006
    对照组 33 25.20(16.30~38.45) 23.70(15.10~33.00) -2.207 0.027 35.20(29.95~40.05) 36.20(33.50~40.00) -2.472 0.019
    Z -0.020 -2.165 0.326 1.760
    P 0.984 0.030 0.745 0.083
    下载: 导出CSV

    表  3  两组T淋巴细胞亚群水平比较

    Table  3.   Comparison of T lymphocyte subsets between the two groups

    组别 例数 CD3+T(%) CD4+T(%)
    治疗前 治疗后 Z P 治疗前 治疗后 Z P
    观察组 32 61.10(49.73~68.80) 71.55(67.60~74.25) -5.189 < 0.001 32.05(24.48~40.28) 43.20(38.70~46.15) -5.321 < 0.001
    对照组 33 61.70(48.90~68.70) 68.20(60.35~71.10) -6.745 < 0.001 33.70(25.85~39.95) 38.00(32.85~41.45) -2.537 0.011
    Z -0.066 -2.146 -0.217 -2.940
    P 0.948 0.032 0.829 0.003
    组别 例数 CD8+T(%) CD4+/CD8+
    治疗前 治疗后 Z P 治疗前 治疗后 Z P
    观察组 32 26.40(22.00~36.75) 25.55(20.65~28.43) 2.565 0.015 1.27(0.92~1.52) 1.69(1.55~1.93) -6.983 < 0.001
    对照组 33 29.70(24.25~36.25) 27.70(23.50~30.90) 1.788 0.083 1.05(0.72~1.59) 1.43(1.06~1.80) -2.381 0.023
    Z -0.513 -1.887 -0.741 3.157
    P 0.610 0.064 0.458 0.002
    下载: 导出CSV

    表  4  两组治疗前后炎症指标比较

    Table  4.   Comparison of inflammatory indexes between the two groups

    组别 例数 IL-6(pg/mL) TNFα(pg/mL) ET(pg/mL)
    治疗前 治疗后 治疗前 治疗后 治疗前 治疗后
    观察组 32 5.04(4.05~6.00) 2.73(1.77~3.16) 57.59(42.37~67.27) 26.94(16.20~42.59) 9.28(7.49~11.41) 6.18(4.00~8.20)
    对照组 33 4.99(4.27~6.18) 3.05(2.51~3.90) 58.38(32.30~71.68) 48.18(15.69~65.41) 10.05(8.59~12.16) 8.12(5.48~9.90)
    Z -0.518 -2.139 -0.039 -1.982 -0.688 -2.062
    P 0.604 0.032 0.969 0.048 0.494 0.043
    下载: 导出CSV
  • [1] SANTIAGO A, POZUELO M, POCA M, et al. Alteration of the serum microbiome composition in cirrhotic patients with ascites[J]. Sci Rep, 2016, 6: 25001. DOI: 10.1038/srep25001.
    [2] MUÑOZ L, BORRERO MJ, U ' BEDA M, et al. Intestinal immune dysregulation driven by dysbiosis promotes barrier disruption and bacterial translocation in rats with cirrhosis[J]. Hepatology, 2019, 70(3): 925-938. DOI: 10.1002/hep.30349.
    [3] LIU JK, ZHAO WX, LIU QH, et al. Effect of Jiedu Huayu Tongfu Recipe on endotoxin and inflammatory factors in patients with alcoholic liver cirrhosis[J]. China J Tradit Chin Med Pharma, 2017, 32(12): 5668-5671. https://www.cnki.com.cn/Article/CJFDTOTAL-BXYY201712115.htm

    刘江凯, 赵文霞, 刘巧红, 等. 解毒化瘀通腑方对酒精性肝硬化患者内毒素及炎性因子的影响[J]. 中华中医药杂志, 2017, 32(12): 5668-5671. https://www.cnki.com.cn/Article/CJFDTOTAL-BXYY201712115.htm
    [4] Chinese Society of Infectious Diseases, Chinese Medical Association, Chinese Society of Hepatology, Chinese Medical Association. Guidelines for the prevention and treatment of chronic hepatitis B (version 2019)[J]. J Clin Hepatol, 2019, 35(12): 2648-2669. DOI: 10.3969/j.issn.1001-5256.2019.12.007.

    中华医学会感染病学分会, 中华医学会肝病学分会. 慢性乙型肝炎防治指南(2019年版)[J]. 临床肝胆病杂志, 2019, 35(12): 2648-2669. DOI: 10.3969/j.issn.1001-5256.2019.12.007.
    [5] Branch of Hepatobiliary Diseases, Chinese Association of Chinese Medicine. The standards of traditional Chinese medicine syndrome differentiation for viral hepatitis[J]. J Clin Hepatol, 2017, 33(10): 1839-1846. DOI: 10.3969/j.issn.1001-5256.2017.10.002.

    中华中医药学会肝胆病分会. 病毒性肝炎中医辨证标准[J]. 临床肝胆病杂志, 2017, 33(10): 1839-1846. DOI: 10.3969/j.issn.1001-5256.2017.10.002.
    [6] ZHENG XY. Guiding principles for clinical research of new traditional Chinese medicine (Trial)[M]. Beijing: China Medical Science and Technology Press, 2002.

    郑筱萸. 中药新药临床研究指导原则(试行)[M]. 北京: 中国医药科技出版社, 2002.
    [7] MINEMURA M, SHIMIZU Y. Gut microbiota and liver diseases[J]. World J Gastroenterol, 2015, 21(6): 1691-1702. DOI: 10.3748/wjg.v21.i6.1691.
    [8] WANG WF, CAO JB, FAN GR, et al. Meta analysis of probiotics in patients with liver cirrhosis[J]. Beijing Med J, 2015, 37 (3): 213-219. DOI: 10.15932/j.0253-9713.2015.3.005.

    王伟芳, 曹建彪, 范公忍, 等. 益生菌在肝硬化患者应用的Meta分析[J]. 北京医学, 2015, 37(3): 213 -219. DOI: 10.15932/j.0253-9713.2015.3.005.
    [9] SOLÉ C, GUILLY S, DA SILVA K, et al. Alterations in gut microbiome in cirrhosis as assessed by quantitative metagenomics: Relationship with acute-on-chronic liver failure and prognosis[J]. Gastroenterology, 2021, 160(1): 206-218. e13. DOI: 10.1053/j.gastro.2020.08.054.
    [10] MARTINS M, BATISTA A, BRITO Y, et al. Effect of remote ischemic preconditioning on systemic toxicity and ototoxicity induced by cisplatin in rats: Role of TNF-α and Nitric Oxide[J]. ORL J Otorhinolaryngol Relat Spec, 2017, 79(6): 336-346. DOI: 10.1159/000485514.
    [11] ZHANG W, PENG Q. Relationship between intestinal flora and liver function and serum inflammatory factors in patients with hepatitis B cirrhosis[J]. Mod Digest Interv, 2020, 25(2): 158-162. DOI: 10.3969/j.issn.1672-2159.2020.02.006.

    张雯, 彭琼. 乙肝肝硬化患者肠道菌群与肝功能及血清炎症因子水平的关系[J]. 现代消化及介入诊疗, 2020, 25(2): 158-162. DOI: 10.3969/j.issn.1672-2159.2020.02.006.
    [12] LARIO M, MUÑOZ L, UBEDA M, et al. Defective thymopoiesis and poor peripheral homeostatic replenishment of T-helper cells cause T-cell lymphopenia in cirrhosis[J]. J Hepatol, 2013, 59(4): 723-730. DOI: 10.1016/j.jhep.2013.05.042.
    [13] MUNOZ L, ALBILLOS A, NIETO M, et al. Mesenteric Th1 polarization and monocyte TNF-alpha production: First steps to systemic inflammation in rats with cirrhosis[J]. Hepatology, 2005, 42(2): 411-419.
    [14] MCGOVERN BH, GOLAN Y, LOPEZ M, et al. The impact of cirrhosis on CD4+ T cell counts in HIV-seronegative patients[J]. Clin Infect Dis, 2007, 44(3): 431-437. DOI: 10.1086/509580.
    [15] ZENG Y, CHEN S, FU Y, et al. Gut microbiota dysbiosis in patients with hepatitis B virus-induced chronic liver disease covering chronic hepatitis, liver cirrhosis and hepatocellular carcinoma[J]. J Viral Hepat, 2020, 27(2): 143-155. DOI: 10.1111/jvh.13216.
    [16] GUO XX, HU N, LIAN XX, et al. Features of intestinal flora imbalance in patients with liver cirrhosis and related driving factors[J]. J Clin Hepatol, 2020, 36(7): 1527-1533. DOI: 10.3969/j.issn.1001-5256.2020.07.016.

    郭晓霞, 胡娜, 廉晓晓, 等. 肝硬化患者肠道菌群失调的特征及驱动因子分析[J]. 临床肝胆病杂志, 2020, 36(7): 1527-1533. DOI: 10.3969/j.issn.1001-5256.2020.07.016.
    [17] ZENG Y, CHEN S, FU Y, et al. Gut microbiota dysbiosis in patients with hepatitis B virus-induced chronic liver disease covering chronic hepatitis, liver cirrhosis and hepatocellular carcinoma[J]. J Viral Hepat, 2020, 27(2): 143-155.
    [18] ABENAVOLI L, SCARPELLINI E, COLICA C, et al. Gut microbiota and obesity: A role for probiotics[J]. Nutrients, 2019, 11(11): 2690. DOI: 10.3390/nu11112690.
    [19] SHEN ZH, ZHU CX, QUAN YS, et al. Relationship between intestinal microbiota and ulcerative colitis: Mechanisms and clinical application of probiotics and fecal microbiota transplantation[J]. World J Gastroenterol, 2018, 24(1): 5-14. DOI: 10.3748/wjg.v24.i1.5.
    [20] CAO H, WU DS, ZHANG Y, et al. Effects of shaoyao decoction on intestinal flora of ulcerative colitis rats based on high-throughput sequencing technology[J]. Chin J Inf Tradit Chin Med, 2021, 28(1): 61-66. DOI: 10.19879/j.cnki.1005-5304.202006249.

    曹晖, 吴东升, 张彧, 等. 基于高通量测序技术研究芍药汤对溃疡性结肠炎大鼠肠道菌群的影响[J]. 中国中医药信息杂志, 2021, 28(1): 61-66. DOI: 10.19879/j.cnki.1005-5304.202006249.
    [21] ZHANG F, LEE J, LIANG S, et al. Cyanobacteria blooms and non-alcoholic liver disease: evidence from a county level ecological study in the United States[J]. Environ Health, 2015, 14: 41. DOI: 10.1186/s12940-015-0026-7.
  • 加载中
图(4) / 表(4)
计量
  • 文章访问数:  364
  • HTML全文浏览量:  81
  • PDF下载量:  40
  • 被引次数: 0
出版历程
  • 收稿日期:  2021-08-25
  • 录用日期:  2021-11-05
  • 出版日期:  2022-04-20
  • 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