药学学报, 2019, 54(4): 670-677
引用本文:
刘佳星, 王彦礼, 李彧, 邹迪新, 王敦方, 马旭冉, 宋红新, 杨伟鹏, 王海南. 四神丸对腹泻型肠易激综合征大鼠肠道菌群影响的实验研究[J]. 药学学报, 2019, 54(4): 670-677.
LIU Jia-xing, WANG Yan-li, LI Yu, ZOU Di-xin, WANG Dun-fang, MA Xu-ran, SONG Hong-xin, YANG Wei-peng, WANG Hai-nan. Experimental study on the effect of Sishen Wan on intestinal flora in rats with diarrhea-type irritable bowel syndrome[J]. Acta Pharmaceutica Sinica, 2019, 54(4): 670-677.

四神丸对腹泻型肠易激综合征大鼠肠道菌群影响的实验研究
刘佳星1, 王彦礼2, 李彧3, 邹迪新2, 王敦方2, 马旭冉2, 宋红新2, 杨伟鹏2, 王海南4
1. 贵州大学药学院, 贵州 贵阳 550025;
2. 中国中医科学院中药研究所, 北京 100700;
3. 北京中医药大学, 北京 100029;
4. 国家药品监督管理局, 北京 100037
摘要:
研究四神丸对腹泻型肠易激综合征(diarrhea-predominant irritable bowel syndrome,IBS-D)大鼠肠道菌群的影响,探讨其改善IBS-D与肠道菌群的关系。选取SPF级雄性SD大鼠45只,随机分为正常组、模型组、四神丸组、二神丸组和五味子散组。以复合因素制备脾肾阳虚型IBS-D大鼠模型后给药14天,采集大鼠粪便,提取粪便样本总DNA,根据细菌16S r RNA V3~V4区设计引物进行扩增,利用Illumina Miseq平台进行高通量测序。动物实验操作过程依照中国中医科学院动物实验伦理委员会的要求执行。研究发现:四神丸能有效减小IBS-D大鼠腹泻指数(P<0.05)及降低肠道高敏感性(P<0.05)。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5组大鼠肠道菌群组成存在明显差异。模型组的菌群丰度、均匀度和多样性均最低。与正常组相比,模型组变形菌门、放线菌门、韦荣球菌属和支原体属显著增加(P<0.05),普雷沃氏菌属显著减少(P<0.05)。与模型组相比,四神丸组变形菌门和支原体属显著减少(P<0.05),梭菌属、Turicibacter和Romboutsia显著增加(P<0.05)。本研究表明四神丸可能是部分通过调节肠道菌群的结构而发挥治疗IBS-D的作用,且二神丸和五味子散之间有协同作用。
关键词:    四神丸      二神丸      五味子散      腹泻型肠易激综合征      肠道菌群      高通量测序     
Experimental study on the effect of Sishen Wan on intestinal flora in rats with diarrhea-type irritable bowel syndrome
LIU Jia-xing1, WANG Yan-li2, LI Yu3, ZOU Di-xin2, WANG Dun-fang2, MA Xu-ran2, SONG Hong-xin2, YANG Wei-peng2, WANG Hai-nan4
1. School of Pharmacy, Guizhou University, Guiyang 550025, China;
2. Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China;
3. Beijing University of Chinese Medicine, Beijing 100029, China;
4. State Drug Administration, Beijing 100037, China
Abstract:
To investigate the effect of Sishen Wan (SSW) on intestinal flora in diarrhea-predominant irritable bowel syndrome (IBS-D) rats and explore the efficacy of this regiment for improving IBS-D, we divided 45 SPF male SD rats randomly into control, disease, SSW, Ershen Wan (ESW) and Wuweizasan (WWZS) groups. The spleen-kidney-yang deficiency type IBS-D rat model was prepared by a composite factor and administered for 14 days. After collecting the feces of the rats, total DNA was extracted from the stool samples. Primers were designed based on the 16S r RNA V3 to V4 regions of the bacteria, and used for high-throughput sequencing with the Illumina Miseq platform. We found that SSW can effectively reduce the diarrhea index (P<0.05) and reduce the high sensitivity of intestinal tract (P<0.05) of IBS-D rats. The principal component analysis (PCA), principal co-ordinates analysis (PCoA) and 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 five groups (P<0.05). The disease group has the lowest in abundance, uniformity and diversity of gut microbiota. Compared with the control group, the disease group showed a significant increase in Proteobacteria, Actinobacteria, Veillonococcus and Mycoplasma (P<0.05), but a significant reduction in Pleaverella (P<0.05). Compared with the disease group, SSW administration caused significant reduction in the Proteobacteria and Mycoplasma (P<0.05), but significant increases of Clostridium, Turicibacter and Romboutsia (P<0.05). Our study shows that SSW has the potential as a therapeutic regiment for treatment of IBS-D due to partial regulation of the intestinal flora. In addition, there is a synergy between ESW and WWZS.
Key words:    Sishen Wan    Er Shen Wan    Wuweizisan    diarrhea-predominant irritable bowel syndrome    intestinal flora    high-throughput sequencing   
收稿日期: 2018-10-11
DOI: 10.16438/j.0513-4870.2018-0920
基金项目: 国家自然科学基金面上项目(81273662,81473592);国家重大新药创制专项资助项目(2014ZX09304306-006);中央级公益性科研院所基本科研业务费专项资金资助(ZXKT17029).
通讯作者: 杨伟鹏, 王海南
Email: hrbywp@sina.com;md_wanghainan@126.com
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参考文献:
[1] Drossman DA. Functional gastrointestinal disorders:history, pathophysiology, clinical features, and Rome IV[J]. Gastroenterology, 2016, 150:1262-1279. e2.
[2] Lacy BE, Mearin F, Chang L, et al. Bowel disorders[J]. Gastroenterology, 2016, 150:1393-1407. e5.
[3] Quigley EM, Fried M, Gwee KA, et al. World gastroenterology organisation global guidelines irritable bowel syndrome:a global perspective update september 2015[J]. J Clin Gastroenterol, 2016, 50:704-713.
[4] Chen F, Zhou XL. Efficacy of flupentixol and melitracen combined with pinaverium bromide in irritable bowel syndrome[J]. Chin J Biochem Pharm (中国生化药物杂志), 2016, 36:86-88.
[5] Xu HY, Wang YL, Wang DF, et al. Effect of Huangqin Tang on the gut microbiota in rats with ulcerative colitis model determined by high-throughput sequencing[J]. Acta Pharm Sin (药学学报), 2017, 52:1673-1682.
[6] Wang ZH, Kuang WC, Hu JJ, et al. Research of Bifidobacterium lactobacillus Sanlian living bacterium in treating alteration of intestinal flora[J]. Shenzhen J Integrat Tradit Chin West Med (深圳中西医结合杂志), 2016, 26:17-19.
[7] Zhu XW, Liu ZB, Niu WM, et al. Effects of electroacupuncture at ST25 and BL25 in a Sennae-induced rat model of diarrhoea-predominant irritable bowel syndrome[J]. Acupunct Med, 2017, 35:216-223.
[8] Fu XQ, Lin RQ, Wu YM, et al. Biochemical study on models of kidney-YANG deficiency replicated by adenine in rats of chronic renal failure[J]. J Fujian Coll Tradit Chin Med (福建中医学院学报), 2003, 13:22-24.
[9] Zhang CM, Liu L, Wang JH, et al. Treatment of spleen-kidney Yang deficiency syndrome of diarrhea type irritable bowel syndrome[J]. Chin Arch Tradit Chin Med (中华中医药学刊), 2017, 35:1376-1378.
[10] Qu CJ, Lin YR, Xia SJ, et al. Comparative study on immunology of purging action of Radix Sophorae Flavescentis in two types spleen-dificiency model[J]. Liaoning J Tradit Chin Med (辽宁中医杂志), 1999, 26:133-134.
[11] Zhao Y, Luo DN, Chen Y, et al. Dose-effect and time-effect relationship of chronic restraint stress combined with senna extract gavage in inducing diarrhea-predominant irritable bowel syndrome in rats[J]. World Chin J Digestol (世界华人消化杂志), 2017, 25:1360-1367.
[12] Yu YB, Zuo XL, Zhao QJ, et al. Brain-derived neurotrophic factor contributes to abdominal pain in irritable bowel syndrome[J]. Gut, 2012, 61:685-694.
[13] Amato KR, Yeoman CJ, Kent A, et al. Habitat degradation impacts black howler monkey (Alouatta pigra) gastrointestinal microbiomes[J]. ISME J, 2013, 7:1344-1353.
[14] Zhao YP, Tang XD, Bian ZX, et al. Preliminary study of establishing and assessing IBS-D model rats of Gan stagnation and Pi deficiency syndrome[J]. Chin J Integrat Tradit West Med (中国中西医结合杂志), 2013, 33:1507-1514.
[15] Zhou F, Liu DY, Zhao HM. Overview of Sishen Pill in the treatment of gastrointestinal diseases[J]. Jiangxi J Tradit Chin Med (江西中医药), 2011, 42:61-63.
[16] An R, Han PY, Xu QY, et al. Establishment and evaluation of animal model of irritable bowel syndrome[J]. J Xinxiang Med Univ (新乡医学院学报), 2016, 33:434-439.
[17] Biedermann L, Rogler G. The intestinal microbiota:its role in health and disease[J]. Eur J Pediat, 2015, 174:151-167.
[18] Chung CS, Chang PF, Liao CH, et al. Differences of microbiota in small bowel and faeces between irritable bowel syndrome patients and healthy subjects[J]. Scand J Gastroenterol, 2016, 51:410-419.
[19] Fan LZ. The curative effect of pinaverium combined with ataining on diarrhea IBS[J]. J Baotou Med Coll (包头医学院学报), 2017, 33:61-62.
[20] Su DM, Li JX, Liu XP. Study on the efficacy and mechanism of ginger moxibustion in treating diarrhea-predominant irritable bowel syndrome by changes of intestinal flora[J]. Chin J Integrat Tradit West Med Digest (中国中西医结合消化杂志), 2018, 26:148-151.
[21] Derrien M, Van Baarlen P, Hooiveld G, et al. Modulation of mucosal immune response, tolerance, and proliferation in mice colonized by the mucin-degrader Akkermansia muciniphila[J]. Front Microbiol, 2011, 2:166.
[22] Ganesh BP, Klopfleisch R, Loh G, et al. Commensal Akkermansia muciniphila exacerbates gut inflammation in Salmonella typhimurium-infected gnotobiotic mice[J]. PLoS One, 2013, 8:e74963.
[23] Delwiche EA, Pestka JJ, Tortorello ML. The veillonellae:gram-negative Cocci with a unique physiology[J]. Annu Rev Microbiol, 1985, 39:175-193.
[24] Matera G, Muto V, Vinci M, et al. Receptor recognition of and immune intracellular pathways for Veillonella parvula lipopolysaccharide[J]. Clin Vaccine Immunol, 2009, 16:1804-1809.
[25] Sommer F, Bäckhed F. The gut microbiota——masters of host development and physiology[J]. Nat Rev Microbiol, 2013, 11:227-238.
[26] Postgate JR. The Sulphate-Reducing Bacteria[M]. Cambridge:Cambridge University Press, 1979.
[27] Gibson GR. Physiology and ecology of the sulphate-reducing bacteria[J]. J Appl Bacteriol, 1990, 69:769-797.
[28] Gibson GR, Macfarlane GT, Cummings JH. Occurrence of sulphate-reducing bacteria in human faeces and the relationship of dissimilatory sulphate reduction to methanogenesis in the large gut[J]. J Appl Bacteriol, 1988, 65:103-111.
[29] Liu H, Yang QJ. Hyperbaric oxygen combined therapy for gas gangrene in 60 cases[J]. Chin J Nosocomiol (中华医院感染学杂志), 2006, 16:1127-1128.
[30] Kelly D,Campbell JI, King TP, et al. Commensal anaerobic gut bacteria attenuate inflammation by regulating nuclear-cytoplasmic shuttling of PPAR-γ and RelA[J]. Nat Immunol, 2004, 5:104-112.
[31] Duncan SH, Louis P, Flint HJ. Lactate-utilizing bacteria, isolated from human feces, that produce butyrate as a major fermentation product[J]. Appl Environ Microbiol, 2004, 70:5810-5817.
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