药学学报, 2017, 52(11): 1673-1682
引用本文:
徐航宇, 王彦礼, 王敦方, 马旭冉, 李洪祥, 杨伟鹏. 高通量测序技术研究黄芩汤对溃疡性结肠炎大鼠肠道菌群的影响[J]. 药学学报, 2017, 52(11): 1673-1682.
XU Hang-yu, WANG Yan-li, WANG Dun-fang, MA Xu-ran, LI Hong-xiang, YANG Wei-peng. Effect of Huangqin Tang on the gut microbiota in rats with ulcerative colitis model determined by high-throughput sequencing [J]. Acta Pharmaceutica Sinica, 2017, 52(11): 1673-1682.

高通量测序技术研究黄芩汤对溃疡性结肠炎大鼠肠道菌群的影响
徐航宇1, 王彦礼1, 王敦方1, 马旭冉1, 李洪祥2, 杨伟鹏1
1. 中国中医科学院中药研究所, 北京 100700;
2. 中国医学科学院、北京协和医学院药用植物研究所, 北京 100193
摘要:
研究黄芩汤(HQT)对溃疡性结肠炎(ulcerative colitis,UC)大鼠肠道菌群的影响,探讨黄芩汤改善溃疡性结肠炎与肠道菌群的关系。选取15只雄性Wistar大鼠随机平均分为对照组(control group)、模型组[三硝基苯磺酸(TNBS) group]和黄芩汤给药组(TNBS+HQT group),以复合法(TNBS+乙醇)制备细胞免疫反应UC大鼠模型。给药10天后,采集15只大鼠粪便样本,提取粪便样本总DNA,根据细菌16S rRNA V3~V4区设计引物进行扩增,利用Illumina Miseq平台进行高通量测序。研究发现:主成分分析(PCA)、主坐标分析(PCoA)和基于β多样性距离的非度量多维尺度分析(NMDS)结果显示3组大鼠肠道菌群组成存在明显差异;与对照组相比,模型组乳酸杆菌属显著减少(P < 0.05),毛螺菌属、脱硫弧菌属、罗氏菌属和瘤胃菌属显著增多(P < 0.05);与模型组比较,黄芩汤给药组乳酸杆菌属显著增多(P < 0.01)、理研菌属显著减少(P < 0.05)。本研究表明黄芩汤可能是部分通过调节肠道菌群的结构而发挥治疗溃疡性结肠炎的作用。
关键词:    黄芩汤      溃疡性结肠炎      肠道菌群      高通量测序     
Effect of Huangqin Tang on the gut microbiota in rats with ulcerative colitis model determined by high-throughput sequencing
XU Hang-yu1, WANG Yan-li1, WANG Dun-fang1, MA Xu-ran1, LI Hong-xiang2, YANG Wei-peng1
1. Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China;
2. Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China
Abstract:
This study was designed to investigate the effect of Huangqin Tang (HQT) on gut microbiota of ulcerative colitis (UC) rats, and to explore the relationship between Huangqin Tang and ulcerative colitis and gut microbiota. Fifteen male Wistar rats were randomly divided into control group, trinitrobenzene sulfonic acid (TNBS) group and TNBS + HQT group. The model of UC rats with cell immunoreactivity was made established using the compound method (TNBS and ethanol). After 10 days of administration, 15 fecal samples were collected and total DNA was extracted from the samples to get total DNA. The primers were designed on bacterial 16S rRNA V3-V4 region sequences and Illumina Miseq platform was used for high-throughput sequencing. It was found that the principal component analysis (PCA), the principal co-ordinates analysis (PCoA) and the non-metric multidimensional scale analysis (NMDS) based on the Beta diversity distance showed that there were significant differences in the composition of the gut microbiota among the three groups (P < 0.05). Compared with the control group, the Lactobacillus of the TNBS group was significantly decreased (P < 0.05), and the Lachnospiraceae, Desulfovibrio, Roseburia, Ruminococcaceae were significantly increased (P < 0.05). Compared with the TNBS group, the Lactobacillus in the TNBS + HQT group was significantly increased (P < 0.01), and the Alistipes was significantly decreased (P < 0.05). The study suggests that the Huangqin Tang plays a role in the treatment of ulcerative colitis partially through regulating the structure of the gut microbiota.
Key words:    Hangqin Tang    ulcerative colitis    gut microbiota    high-throughput sequencing   
收稿日期: 2017-05-21
DOI: 10.16438/j.0513-4870.2017-0512
基金项目: 国家自然科学基金资助项目(81273662,81473592).
通讯作者: 杨伟鹏
Email: hrbywp@sina.com
相关功能
PDF(671KB) Free
打印本文
0
作者相关文章
徐航宇  在本刊中的所有文章
王彦礼  在本刊中的所有文章
王敦方  在本刊中的所有文章
马旭冉  在本刊中的所有文章
李洪祥  在本刊中的所有文章
杨伟鹏  在本刊中的所有文章

参考文献:
[1] Shen H. Ulcerative Colitis:the Past, Present and Future of Chinese and Western Medicine (溃疡性结肠炎:中西医的过去、现在与未来)[M]. Nanjing:Southeast University Press, 2012:29-34.
[2] Yin YN, Wang CL, Liu XW, et al. Gastric and duodenum microflora analysis after long-term Helicobacter pylori infection in Mongolian Gerbils[J]. Helicobacter, 2011, 16:389-397.
[3] Ordás I, Eckmann L, Talamini M, et al. Ulcerative colitis[J]. Lancet, 2012, 380:1606-1619.
[4] Wang ZJ, Chen YP, Huang JY, et al. Effect of colonning on intestinal microecology in patients with ulcerative colitis[J]. Guandong J Med (广东医学), 2016, 37:2033-2035.
[5] Lin Z, Zu XP, Xie HS, et al. Research progress in mechanism of intestinal microorganisms in human diseases[J]. Acta Pharm Sin (药学学报), 2016, 51:843-852.
[6] Scanlan PD, Shanahan F, O'Mahony C, et al. Culture-independent analyses of temporal variation of the dominant fecal microbiota and targeted bacterial subgroups in Crohn's disease[J]. J Clini Micro, 2006, 44:3980-3988.
[7] Yu XD, Lü XZ, Dong WB. Clinical observation on treatment of ulcerative colitis with Huangqin decoction[J]. J Emerg Tradit Chin Med (中国中医急症), 2010, 19:1510
[8] Gu ZJ. Investigation of the Therapeutical Mechanisms on the Ulcerative Colitis by Rheum Tanguticum Polysaccharide (大黄多糖治疗溃疡性结肠炎的机制研究)[D]. Xi'an:Fourth Military Medical University, 2013.
[9] Wang DF, Wang YL, Wang YW, et al. Effect of Huangqin Tang on the function of regulatory TLR4/My D88 signal pathway in rats with ulcerative colitis[J]. Acta Pharm Sin (药学学报), 2016, 51:1558-1563.
[10] Wang YW, Zhang HH, Wang YL, et al. Effect of Huangqin Tang on the regulatory NF-κB p65 signal pathway in rats with ulcerative colitis[J]. Acta Pharm Sin (药学学报), 2015, 50:21-27.
[11] Zhuang SX. Study on the Antioxidative Stress of Huangqin Tang on Ulcerative Colitis Through the Nrf2 Pathway (黄芩汤调控Nrf2通路对溃疡性结肠炎的抗氧化应激作用机制研究)[D]. Beijing:China Academy of Chinese Medical Sciences, 2016.
[12] Tao L, Zhuang S, Wang Y, et al. Flavonoid profiling of a traditional Chinese medicine formula of Huangqin Tang using high performance liquid chromatography[J]. Acta Pharm Sin B, 2016, 6:148-157.
[13] Morris GP, Beck PL, Herridge MS, et al. Hapten-induced model of chronic inflammation and ulceration in the rat co-lon[J]. Gastroenterology, 1989, 96:795-803.
[14] Cooper HS, Murthy SN, Shah RS, et al. Clinicopathologic study of dextran sulfate sodium experimental murine colitis[J]. Lab Invest, 1993, 69:238-249.
[15] Amato KR, Yeoman CJ, Kent A, et al. Habitat degradation impacts black howler monkey (Alouatta pigra) ga-strointestinal microbiomes[J]. ISME J, 2013, 7:1344-1353.
[16] Wang Y, Sheng HF, He Y, et al. Comparison of the levels of bacterial diversity in freshwater, intertidal wetland, and marine sediments by using millions of illumina tags[J]. Appl Environ Microbiol, 2012, 78:8264-8271.
[17] Bel S, Elkis Y, Elifantz H, et al. Reprogrammed and trans-missible intestinal microbiota confer diminished susceptibility to induced colitis in TMF-/- mice[J]. Proc Nat Acad Sci U S A, 2014, 111:4964-4969.
[18] Vlantis K, Polykratis A, Welz PS, et al. Original article:TLR-independent anti-inflammatory function of intestinal epithelial TRAF6 signalling prevents DSS-induced colitis in mice[J]. Gut, 2016, 65:935-943.
[19] Annese V, Daperno M, Rutter MD, et al. European evidence based consensus for endoscopy in inflammatory bowel disease[J]. J Crohns Colitis, 2013, 7:982-1018.
[20] Panes J, Bouhnik Y, Reinisch W, et al. Imaging techniques for assessment of inflammatory bowel disease:joint ECCO and ESGAR evidence-based consensus guidelines[J]. J Crohns Colitis, 2013, 7:556-585.
[21] Salminen S, Bouley C, Boutron MC, et al. Functional food science and gastrointestinal physiology and function[J]. Br J Nutr, 1998, 80:147-171.
[22] Sanz Y, Nadal I, Sánchez E. Probiotics as drugs against human gastrointestinal infections[J]. Recent Pat Anticancer Drug Discov, 2007, 2:148-156.
[23] Bernet MF, Brassart D, Neeser JR, et al. Lactobacillus acidophilus LA 1 binds to cultured human intestinal cell lines and inhibits cell attachment and cell invasion by enterovirulent bacteria[J]. Gut, 1994, 35:483-489.
[24] Ljungh A, Wadström T. Lactic acid bacteria as probiotics[J]. Cur Issues Intes Micro, 2006, 7:73-90.
[25] Galdeano CM, Perdigon G. The probiotic bacterium Lacto-bacillus casei induces activation of the gut mucosal immune system through innate immunity[J]. Clin Vaccine Immunol, 2006, 13:219-226.
[26] Chen K. Analysis of Sulfate-Reducing Bacteria in the Gut of Ulcerative Colitis Patients (溃疡性结肠炎患者肠道硫酸盐还原菌的检测)[D]. Shanghai:Shanghai Jiao Tong University, 2014.
[27] Gao XJ, Li T, Wei B, et al. Regulatory mechanisms of gut microbiota on intestinal CYP3A and P-glycoprotein in rats with dextran sulfate sodium-induced colitis[J]. Acta Pharm Sin (药学学报), 2017, 52:34-43.
[28] Zhang M, Liu B, Zhang Y, et al. Structural shifts of mu-cosa-associated Lactobacilli and Clostridium leptum subgroup in patients with ulcerative colitis[J]. J Clin Micro, 2007, 45:496-500.
[29] Angelberger S, Reinisch W, Makristathis A, et al. Temporal bacterial community dynamics vary among ulcerative colitis patients after fecal microbiota transplantation[J]. Am J Gastroenterol, 2013, 108:1620-1630.
相关文献:
1.王敦方, 王彦礼, 王怡薇, 杜丽坤, 佟颖, 陈曦, 郭姗姗, 徐航宇, 马旭冉, 李涛, 杨伟鹏.基于UHPLC-MS研究黄芩汤对溃疡性结肠炎大鼠血清代谢谱的影响[J]. 药学学报, 2017,52(8): 1306-1312
2.王敦方, 王彦礼, 王怡薇, 郭姗姗, 庄帅星, 徐航宇, 李涛, 杨伟鹏.黄芩汤对溃疡性结肠炎大鼠TLR4/MyD88通路调控作用研究[J]. 药学学报, 2016,51(10): 1558-1563
3.王怡薇, 张会会, 王彦礼, 郭姗姗, 李涛, 陈立, 庄帅星, 周钟鸣, 杨伟鹏.黄芩汤对溃疡性结肠炎大鼠NF-κBp65调控作用研究[J]. 药学学报, 2015,50(1): 21-27