人参中性多糖对小鼠肠道菌群组成及多样性的影响
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摘要
探讨人参多糖中的中性多糖级分对抗生素相关性腹泻(antibiotic associated diarrhea,AAD)小鼠的治疗作用及对肠道菌群组成和多样性的影响。通过水提、醇沉,从人参根中得到水溶性人参多糖(WGP),再通过离子交换色谱纯化和制备人参中性多糖(WGPN)。利用灌胃盐酸林可霉素建立AAD小鼠模型,随后灌胃生理盐水(自然恢复组,NR)或给药WGPN(WGPN组) 1周,观察并记录小鼠体重变化、精神状态和腹泻情况。末次给药12 h后,收集小鼠肠道内容物进行16S rRNA高通量测序,并取回肠进行组织学检查。结果表明,WGPN能够减轻腹泻小鼠症状,改善小鼠回肠组织水肿和炎症情况,增加肠绒毛长度。与NR组相比,属水平上,WGPN能够增加乳杆菌属相对丰度,显著降低拟杆菌属、链球菌属、苍白杆菌属和假单胞菌属的相对丰度。因此,WGPN可通过促进AAD小鼠肠道结构修复,调节小鼠肠道菌群的组成和多样性,达到重建肠道微生态的作用。
To investigate the effect of ginseng neutral polysaccharide on gut microbiota composition and diversity as well as the therapeutic effect for antibiotic associated diarrhea( AAD) in mice. The water-soluble ginseng neutral polysaccharide( WGPN) was purified from water-soluble ginseng polysaccharides( WGP) by DEAE-sepharose fast flow column,which was obtained from the roots of Panax ginseng. AAD mice were induced by gastric gavage with lincomycin hydrochloride,followed by administration of normal saline( natural recovery group,NR) or WGPN( WGPN group) for one week. Body weight changes,psychosis and diarrhea status were observed and assessed. 12 h after the last administration,histological observation of ileum and 16 S rRNA high throughput sequencing analysis of intestinal contents were conducted to identify the effects of WGPN on AAD mice. The results showed that WGPN could alleviate the symptoms of diarrhea in mice,decrease the inflammation and edema of ileum,and increase the length of intestinal villi. As compared to NR mice,WGPN could increase the relative abundance of Lactobacillus,and significantly decrease the relative abundance of Bacteroides,Streptococcus,Ochrobactrum and Pseudomonas at the genus level. In conclusion,WGPN could improve the gut microecology by recovering the ileum structure and improving the diversity and composition of the gut microbiota in AAD mice.
To investigate the effect of ginseng neutral polysaccharide on gut microbiota composition and diversity as well as the therapeutic effect for antibiotic associated diarrhea( AAD) in mice. The water-soluble ginseng neutral polysaccharide( WGPN) was purified from water-soluble ginseng polysaccharides( WGP) by DEAE-sepharose fast flow column,which was obtained from the roots of Panax ginseng. AAD mice were induced by gastric gavage with lincomycin hydrochloride,followed by administration of normal saline( natural recovery group,NR) or WGPN( WGPN group) for one week. Body weight changes,psychosis and diarrhea status were observed and assessed. 12 h after the last administration,histological observation of ileum and 16 S rRNA high throughput sequencing analysis of intestinal contents were conducted to identify the effects of WGPN on AAD mice. The results showed that WGPN could alleviate the symptoms of diarrhea in mice,decrease the inflammation and edema of ileum,and increase the length of intestinal villi. As compared to NR mice,WGPN could increase the relative abundance of Lactobacillus,and significantly decrease the relative abundance of Bacteroides,Streptococcus,Ochrobactrum and Pseudomonas at the genus level. In conclusion,WGPN could improve the gut microecology by recovering the ileum structure and improving the diversity and composition of the gut microbiota in AAD mice.
引文
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[35] Xu X,Xu P,Ma C,et al. Gut microbiota,host health,and pol-ysaccharides[J]. Biotechnol Adv,2013,31(2):318.
[2] Sampson T R,Debelius J W,Thron T,et al. Gut microbiota reg-ulate motor deficits and neuroinflammation in a model of Parkin-son's disease[J]. Cell,2016,167(6):1469.
[3] Thomas S,Izard J,Walsh E,et al. The host microbiome regu-lates and maintains human health:a primer and perspective fornon-microbiologists[J]. Cancer Res,2017,77(8):1783.
[4] Marchesi J R,Adams D H,Fava F,et al. The gut microbiotaand host health:a new clinical frontier[J]. Gut,2016,65(2):330.
[5] Bartlett J G. Clinical practice. Antibiotic-associated diarrhea[J]. N Engl J Med,2002,346(5):334.
[6] Wong S,Santullo P,Hirani S P,et al. Use of antibiotics and theprevalence of antibiotic-associated diarrhoea in patients with spi-nal cord injuries:an international,multicentre centre study[J].J Hosp Infect,2017,97(2):146.
[7] Lv W,Liu C,Ye C,et al. Structural modulation of gut microbio-ta during alleviation of antibiotic-associated diarrhea with herbalformula[J]. Int J Biol Macromol,2017,105(3):1622.
[8] Akhter K F,Mumin M A,Lui E M K,et al. Fabrication of fluo-rescent labeled ginseng polysaccharide nanoparticles for bioimag-ing and their immunomodulatory activity on macrophage cell lines[J]. Int J Biol Macromol,2018,109(1):254.
[9] Zhang X,Li S,Sun L,et al. Further analysis of the structureand immunological activity of an RG-I type pectin from Panaxginseng[J]. Carbohydr Polym,2012,89(2):519.
[10] Cheng H,Li S,Fan Y,et al. Comparative studies of the antipro-liferative effects of ginseng polysaccharides on HT-29 human co-lon cancer cells[J]. Med Oncol,2011,28(1):175.
[11]杨蕾,张振海,贾晓斌.人参稀有皂苷组分联合紫杉醇治疗A549肺癌的实验研究[J].中国中药杂志,2018,43(7):1446.
[12] Wang J,Flaisher-Grinberg S,Li S,et al. Antidepressant-likeeffects of the active acidic polysaccharide portion of ginseng inmice[J]. J Ethnopharmacol,2010,132(1):65.
[13]刘丽琴,罗艳,张瑞睿,等.人参皂苷对慢性应激抑郁模型大鼠行为学及HPA轴、BDNF的影响[J].中国中药杂志,2018,36(10):1342.
[14] Wang J,Li S,Fan Y,et al. Anti-fatigue activity of the water-soluble polysaccharides isolated from Panax ginseng C. A. Meyer[J]. J Ethnopharmacol,2010,130(2):421.
[15] Zhou S S,Xu J,Zhu H,et al. Gut microbiota-involved mecha-nisms in enhancing systemic exposure of ginsenosides by coexis-ting polysaccharides in ginseng decoction[J]. Sci Rep,2016,6:22474.
[16] Zhang X,Yu L,Bi H,et al. Total fractionation and characteriza-tion of the water-soluble polysaccharides isolated from Panaxginseng C. A. Meyer[J]. Carbohydr Polym,2009,77(3):544.
[17] Fu D,O'Neill R A. Monosaccharide composition analysis of oli-gosaccharides and glycoproteins by high-performance liquid chro-matography[J]. Anal Biochem,1995,227(2):377.
[18]苑鹤,白燕冰,斯金平,等.柱前衍生HPLC分析铁皮石斛多糖中单糖组成的变异规律[J].中国中药杂志,2011,36(18):2465.
[19] Lange K,Buerger M,Stallmach A,et al. Effects of antibiotics ongut microbiota[J]. Dig Dis,2016,34(3):260.
[20] Vangay P,Ward T,Gerber J S,et al. Antibiotics,pediatric dys-biosis and disease[J]. Cell Host Microbe,2015,17(5):553.
[21] Caspary W F. Physiology and pathophysiology of intestinal ab-sorption[J]. Am J Clin Nutr,1992,55(1 Suppl):299S.
[22] Johnsonhenry K C,Mitchell D J,Avitzur Y,et al. Probiotics re-duce bacterial colonization and gastric inflammation in H. pylori-infected mice[J]. Dig Dis Sci,2004,49(7/8):1095.
[23] Bergognebérézin E. Treatment and prevention of antibiotic associ-ated diarrhea[J]. Int J Antimicrob Agents,2013,16(4):521.
[24] Lin Y,Vonk R J,Slooff M J,et al. Differences in propionate-induced inhibition of cholesterol and triacylglycerol synthesis be-tween human and rat hepatocytes in primary culture[J]. Br JNutr,1995,74(2):197.
[25] Wang Y,Wu Y,Wang Y,et al. Antioxidant properties of probi-otic bacteria[J]. Nutrients,2017,9(5):521.
[26] Choi S,Kim Y,Han K,et al. Effects of Lactobacillus strains oncancer cell proliferation and oxidative stress in vitro[J]. Lett Ap-pl Microbiol,2006,42(5):452.
[27] Wang C,Zhu Y,Li F,et al. The effect of Lactobacillus isolateson growth performance,immune response,intestinal bacterialcommunity composition of growing rex rabbits[J]. J Anim Physi-ol Anim Nutr(Berl),2017,101(5):1.
[28] Petri D,Hill J E,Kessel A G V,et al. Microbial succession inthe gastrointestinal tract(GIT)of the preweaned pig[J]. LivestSci,2010,133(1):107.
[29]祁玉丽,高坤,孙印石,等.植物多糖对肠道微生态的作用研究进展[J].中国微生态学杂志,2018(4):489.
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[31]黄静,李胜立,赵宏伟.霍山石斛多糖对四氯化碳致急性肝损伤小鼠的保护作用[J].中国中药杂志,2013,38(4):528.
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