四君子汤及其单味药水煎液对脾虚大鼠肠道菌群的调节作用
|
摘要
目的:探讨四君子汤及其单味药(人参、白术、茯苓、炙甘草)水煎液对脾虚大鼠肠道菌群的调节作用。方法:正常大鼠随机分为空白组、模型组、整肠生颗粒组、四君子汤组及各单味药组,连续10 d灌胃给予番泻叶水煎液塑造脾虚模型(空白组给予蒸馏水),再连续7 d分别给予相应药物治疗(空白组、模型组给予蒸馏水)。收集造模前(第0天),造模结束(第11天)和治疗结束(第18天)的粪便样品,经硫酸酸化、乙醚萃取处理后采用气相色谱法分析样品中短链脂肪酸含量的变化,利用16S rDNA-聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术分析粪便样品中肠道菌群结构的变化。结果:造模结束时,与空白组比较,各给药组大鼠粪便短链脂肪酸含量、肠道菌群多样性指数和相似性系数均显著下降(P <0. 05,P <0. 01),表明脾虚模型造模成功。治疗结束时,与模型组比较,白术组大鼠粪便短链脂肪酸含量、肠道菌群多样性指数和相似性系数均极显著升高(P <0. 01),四君子汤组、人参组和茯苓组部分指标极显著升高、部分指标显著升高(P <0. 05,P <0. 01);而炙甘草组仅多样性指数有所升高(P <0. 05),粪便短链脂肪酸含量和相似性系数则均无显著性差异。结论:四君子汤对脾虚大鼠肠道菌群的恢复调节时,白术可能发挥主要作用,人参、茯苓也发挥了一定作用。
Objective:To investigate the regulatory effect of Sijunzi Tang(SJZT)and its single herbs(Ginseng Radix et Rhizoma,Glycyrrhizae Radix et Rhizoma Praeparata cum Melle,Atractylodis Macrocephalae Rhizoma and Poria)on intestinal flora in spleen-deficient rats.Method:Normal rats were randomly divided into the blank group,model group,Zhengchangsheng granules group,SJZT group and each single herb group,rats were orally administered Sennae Folium decoction to induce diarrhea for ten consecutive days to establish a spleendeficient model(distilled water for the blank group),then treated with the corresponding drugs for seven consecutive days(distilled water for the blank group and the model group).Fresh feces were collected on premodeling(0~(th)day),post-modeling(11~(th)day),and post-treatment(18~(th)day).Short-chain fatty acids(SCFAs)in feces were acidified by sulphuric acid and extracted by diethyl ether,then determined by gas chromatography.The structural change(diversity and similarity)of intestinal flora in feces was analyzed by 16S r DNA-polymerase chain reaction(PCR)-denaturing gel gradient electrophoresis(DGGE)technique.Result:Compared with blank group,the contents of SCFAs as well as diversity and similarity indexes of intestinal flora in feces of all administered groups were significantly decreased on the 11~(th)day(P<0.05,P<0.01),this indicated that the spleen-deficient model was successfully constructed.On the 18~(th)day,compared with model group,the contents of SCFAs as well as diversity and similarity indexes of intestinal flora in feces of Atractylodis Macrocephalae Rhizoma group were significantly increased(P<0.01),these indexes in feces of SJZT group,Ginseng Radix et Rhizoma group and Poria group were significantly increased(P<0.05,P<0.01),while Glycyrrhizae Radix et Rhizoma Praeparata cum Melle group only increased diversity index of intestinal flora(P<0.05).Conclusion:Ginseng Radix et Rhizoma,Atractylodis Macrocephalae Rhizoma and Poria are the single herbs responsible for the regulatory effect of SJZT on intestinal flora in spleen-deficient rats,and Atractylodis Macrocephalae Rhizoma may play a major role.
Objective:To investigate the regulatory effect of Sijunzi Tang(SJZT)and its single herbs(Ginseng Radix et Rhizoma,Glycyrrhizae Radix et Rhizoma Praeparata cum Melle,Atractylodis Macrocephalae Rhizoma and Poria)on intestinal flora in spleen-deficient rats.Method:Normal rats were randomly divided into the blank group,model group,Zhengchangsheng granules group,SJZT group and each single herb group,rats were orally administered Sennae Folium decoction to induce diarrhea for ten consecutive days to establish a spleendeficient model(distilled water for the blank group),then treated with the corresponding drugs for seven consecutive days(distilled water for the blank group and the model group).Fresh feces were collected on premodeling(0~(th)day),post-modeling(11~(th)day),and post-treatment(18~(th)day).Short-chain fatty acids(SCFAs)in feces were acidified by sulphuric acid and extracted by diethyl ether,then determined by gas chromatography.The structural change(diversity and similarity)of intestinal flora in feces was analyzed by 16S r DNA-polymerase chain reaction(PCR)-denaturing gel gradient electrophoresis(DGGE)technique.Result:Compared with blank group,the contents of SCFAs as well as diversity and similarity indexes of intestinal flora in feces of all administered groups were significantly decreased on the 11~(th)day(P<0.05,P<0.01),this indicated that the spleen-deficient model was successfully constructed.On the 18~(th)day,compared with model group,the contents of SCFAs as well as diversity and similarity indexes of intestinal flora in feces of Atractylodis Macrocephalae Rhizoma group were significantly increased(P<0.01),these indexes in feces of SJZT group,Ginseng Radix et Rhizoma group and Poria group were significantly increased(P<0.05,P<0.01),while Glycyrrhizae Radix et Rhizoma Praeparata cum Melle group only increased diversity index of intestinal flora(P<0.05).Conclusion:Ginseng Radix et Rhizoma,Atractylodis Macrocephalae Rhizoma and Poria are the single herbs responsible for the regulatory effect of SJZT on intestinal flora in spleen-deficient rats,and Atractylodis Macrocephalae Rhizoma may play a major role.
引文
[1]巩昌镇,刘一凡,杨佃会,等.四君子汤[M].北京:中国医药科技出版社,2009:3,31.
[2]吴三明,张万岱.脾虚泄泻患者肠道微生态学的初步研究[J].中国中西医结合脾胃杂志,1996,4(4):203-204.
[3]王占国.中医“脾”与消化道正常菌群[J].中国微生态学杂志,1991,3(2):65-68.
[4]马祥雪,王凤云,符竣杰,等.基于肠道菌群的中医健脾方剂作用机制的研究现状与思考[J].中国实验方剂学杂志,2017,23(5):210-215.
[5]Topping D L,Clifton P M. Short-chain fatty acids and human colonic function:roles of resistant starch and nonstarch polysaccharides[J]. Physiol Rev,2001,81(3):1031-1064.
[6]Hold G L,Schwiertz A,Aminov R I,et al.Oligonucleotide probes that detect quantitatively significant groups of butyrate-producing bacteria in human feces[J]. Appl Environ Microbiol,2003,69(7):4320-4324.
[7]Wong J M W,Jenkins D J A. Carbohydrate digestibility and metabolic effects[J]. J Nutr,2007,137(11 Suppl):2539S-2546S.
[8]刘松珍,张雁,张名位,等.肠道短链脂肪酸产生机制及生理功能的研究进展[J].广东农业科学,2013,40(11):99-103.
[9]严梅桢,李志军,谢念祥,等.四君子汤对实验性脾虚小鼠肠道菌群的影响[J].中国微生态学杂志,1989,1(1):40-43.
[10]任光友,张贵林,卢素琳,等.四君子汤对动物肠菌失调及正常胃肠功能的药理研究[J].中成药,2000,22(7):504-506.
[11]任晓风,吴莲波,王伟明,等.四君子汤对脾气虚证调节作用的实验研究[J].中国中医药科技,2005,12(6):359-360.
[12]彭颖,金晶,杨静玉,等. 3种健脾补气方药对脾气虚证大鼠肠道菌群的影响[J].中国中药杂志,2008,33(21):2530-2534.
[13]王卓,彭颖,李晓波.四君子汤对两种脾虚模型大鼠肠道菌群紊乱的影响[J].中国中西医结合杂志,2009,29(9):825-829.
[14]曲长江,林庶如,夏淑杰,等.苦寒泻下两种脾虚模型的免疫学比较研究[J].辽宁中医杂志,1999,26(3):133-134.
[15]沈丽波,钱会南.脾虚模型实验研究方法概述[J].中国中医药信息杂志,2005,12(1):93-95.
[16]萨姆布鲁克J,拉塞尔D W.分子克隆实验指南[M].3版.北京:科学出版社,2002:935.
[17]金晶,彭颖,李晓波.快速提取肠道微生物基因组DNA的方法[J].现代生物医学进展,2007,7(1):100-103.
[18]彭颖.脾虚证肠道菌群基因指纹评价指标及相关药物的筛选方法[D].沈阳:沈阳药科大学,2008.
[19]ZHAO G H,Nyman M,J?nsson J ?. Rapid determination of short-chain fatty acids in colonic contents and faeces of humans and rats by acidified water-extraction and direct-injection gas chromatography[J]. Biomed Chromatogr,2006,20(8):674-682.
[20]贾益群,叶福媛,王双,等.生物样品中短链脂肪酸的快速提取与分析方法[J].实验室研究与探索,2012,31(7):262-264.
[21]杨泽锐,曾桂梅,彭丽华,等.红景天破壁饮片对小鼠肠道菌群影响的初步研究[J].中国中药杂志,2015,40(15):3053-3058.
[22]贾宏信,苏米亚,陈文亮,等.婴儿喂养方式的改变对肠道菌群多样性的影响[J].中国微生态学杂志,2016,28(12):1365-1369.
[23]吴莉娟,孙文,吴丽丽,等.糖耐康对T2DM大鼠ZDF肠道菌群结构的影响[J].中国实验方剂学杂志,2017,23(8):98-104.
[24]周霖,岑泳欣,王立生,等.小檗碱对肠易激综合征大鼠肠道菌群的影响[J].中国微生态学杂志,2018,30(7):772-776.
[25]王玉蕾,郑跃杰.肠道中短链脂肪酸与过敏性疾病关系的研究进展[J].中国微生态学杂志,2013,25(1):104-108.
[26]吴鹏,李慧芬,刘江亭,等.白术及其制剂调整脾虚证肠道菌群紊乱的研究概况[J].山东中医杂志,2015,34(7):564-565.
[27]罗云波,贾波,祝捷,等.白术茯苓汤及水煎液提取分离组分对脾气虚大鼠VIP受体的影响[J].时珍国医国药,2010,21(5):1045-1046.
[28]潘爱珍,易伟民,余晓娟,等.人参总皂苷对脾虚模型大鼠的保护作用研究[J].中国药房,2013,24(39):3682-3684.
[29]曾星,贺毅,周丹,等.甘草对脾虚证消化道黏膜及细胞保护的分子药理机制研究进展[J].时珍国医国药,2010,21(8):2030-2031.
[30]舒青龙,王萍,封勇,等.理中汤对抗生素相关性腹泻模型构建中肠道菌群变化的影响[J].中国实验方剂学杂志,2015,21(21):82-87.
[31]王丽芳,高文远,徐鑫,等.神曲鲜干品组方对食积小鼠胃肠动力及肠道菌群调整的影响[J].中国实验方剂学杂志,2017,23(4):20-24.
[32]张宁,李自辉,赵洪伟,等.寒凝血瘀证大鼠的肠道菌群变化与粪便代谢特征分析[J].中国实验方剂学杂志,2018,24(2):79-85.
[2]吴三明,张万岱.脾虚泄泻患者肠道微生态学的初步研究[J].中国中西医结合脾胃杂志,1996,4(4):203-204.
[3]王占国.中医“脾”与消化道正常菌群[J].中国微生态学杂志,1991,3(2):65-68.
[4]马祥雪,王凤云,符竣杰,等.基于肠道菌群的中医健脾方剂作用机制的研究现状与思考[J].中国实验方剂学杂志,2017,23(5):210-215.
[5]Topping D L,Clifton P M. Short-chain fatty acids and human colonic function:roles of resistant starch and nonstarch polysaccharides[J]. Physiol Rev,2001,81(3):1031-1064.
[6]Hold G L,Schwiertz A,Aminov R I,et al.Oligonucleotide probes that detect quantitatively significant groups of butyrate-producing bacteria in human feces[J]. Appl Environ Microbiol,2003,69(7):4320-4324.
[7]Wong J M W,Jenkins D J A. Carbohydrate digestibility and metabolic effects[J]. J Nutr,2007,137(11 Suppl):2539S-2546S.
[8]刘松珍,张雁,张名位,等.肠道短链脂肪酸产生机制及生理功能的研究进展[J].广东农业科学,2013,40(11):99-103.
[9]严梅桢,李志军,谢念祥,等.四君子汤对实验性脾虚小鼠肠道菌群的影响[J].中国微生态学杂志,1989,1(1):40-43.
[10]任光友,张贵林,卢素琳,等.四君子汤对动物肠菌失调及正常胃肠功能的药理研究[J].中成药,2000,22(7):504-506.
[11]任晓风,吴莲波,王伟明,等.四君子汤对脾气虚证调节作用的实验研究[J].中国中医药科技,2005,12(6):359-360.
[12]彭颖,金晶,杨静玉,等. 3种健脾补气方药对脾气虚证大鼠肠道菌群的影响[J].中国中药杂志,2008,33(21):2530-2534.
[13]王卓,彭颖,李晓波.四君子汤对两种脾虚模型大鼠肠道菌群紊乱的影响[J].中国中西医结合杂志,2009,29(9):825-829.
[14]曲长江,林庶如,夏淑杰,等.苦寒泻下两种脾虚模型的免疫学比较研究[J].辽宁中医杂志,1999,26(3):133-134.
[15]沈丽波,钱会南.脾虚模型实验研究方法概述[J].中国中医药信息杂志,2005,12(1):93-95.
[16]萨姆布鲁克J,拉塞尔D W.分子克隆实验指南[M].3版.北京:科学出版社,2002:935.
[17]金晶,彭颖,李晓波.快速提取肠道微生物基因组DNA的方法[J].现代生物医学进展,2007,7(1):100-103.
[18]彭颖.脾虚证肠道菌群基因指纹评价指标及相关药物的筛选方法[D].沈阳:沈阳药科大学,2008.
[19]ZHAO G H,Nyman M,J?nsson J ?. Rapid determination of short-chain fatty acids in colonic contents and faeces of humans and rats by acidified water-extraction and direct-injection gas chromatography[J]. Biomed Chromatogr,2006,20(8):674-682.
[20]贾益群,叶福媛,王双,等.生物样品中短链脂肪酸的快速提取与分析方法[J].实验室研究与探索,2012,31(7):262-264.
[21]杨泽锐,曾桂梅,彭丽华,等.红景天破壁饮片对小鼠肠道菌群影响的初步研究[J].中国中药杂志,2015,40(15):3053-3058.
[22]贾宏信,苏米亚,陈文亮,等.婴儿喂养方式的改变对肠道菌群多样性的影响[J].中国微生态学杂志,2016,28(12):1365-1369.
[23]吴莉娟,孙文,吴丽丽,等.糖耐康对T2DM大鼠ZDF肠道菌群结构的影响[J].中国实验方剂学杂志,2017,23(8):98-104.
[24]周霖,岑泳欣,王立生,等.小檗碱对肠易激综合征大鼠肠道菌群的影响[J].中国微生态学杂志,2018,30(7):772-776.
[25]王玉蕾,郑跃杰.肠道中短链脂肪酸与过敏性疾病关系的研究进展[J].中国微生态学杂志,2013,25(1):104-108.
[26]吴鹏,李慧芬,刘江亭,等.白术及其制剂调整脾虚证肠道菌群紊乱的研究概况[J].山东中医杂志,2015,34(7):564-565.
[27]罗云波,贾波,祝捷,等.白术茯苓汤及水煎液提取分离组分对脾气虚大鼠VIP受体的影响[J].时珍国医国药,2010,21(5):1045-1046.
[28]潘爱珍,易伟民,余晓娟,等.人参总皂苷对脾虚模型大鼠的保护作用研究[J].中国药房,2013,24(39):3682-3684.
[29]曾星,贺毅,周丹,等.甘草对脾虚证消化道黏膜及细胞保护的分子药理机制研究进展[J].时珍国医国药,2010,21(8):2030-2031.
[30]舒青龙,王萍,封勇,等.理中汤对抗生素相关性腹泻模型构建中肠道菌群变化的影响[J].中国实验方剂学杂志,2015,21(21):82-87.
[31]王丽芳,高文远,徐鑫,等.神曲鲜干品组方对食积小鼠胃肠动力及肠道菌群调整的影响[J].中国实验方剂学杂志,2017,23(4):20-24.
[32]张宁,李自辉,赵洪伟,等.寒凝血瘀证大鼠的肠道菌群变化与粪便代谢特征分析[J].中国实验方剂学杂志,2018,24(2):79-85.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |