黄连解毒汤与复合抗生素对大鼠胃肠道菌群结构的影响
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摘要
为了探讨黄连解毒汤与复合抗生素对大鼠胃肠道菌群微生态的调节作用,将大鼠分为对照组、抗生素组和黄连解毒汤高、中、低剂量组,各组大鼠每天灌胃2 mL对应药物。对照组大鼠灌胃纯化水;抗生素组大鼠第1周灌胃氨苄西林56 mg/mL,第2周灌胃氨苄西林+链霉素(56+28)mg/mL,第3周灌胃氨苄西林+链霉素+克林霉素(56+28+50)mg/mL;黄连解毒汤高、中、低剂量组大鼠分别灌胃黄连解毒汤192,96,48 mg/mL。于给药后第7,14,21天处死大鼠,收集胃、十二指肠、空肠、回肠和结肠的内容物,提取细菌DNA,采用肠杆菌基因间重复一致序列聚合酶链反应(ERIC-PCR)技术对胃肠道各段中细菌菌群的结构多样性进行检测。结果表明:对照组大鼠胃肠道菌群结构相对稳定,ERIC-PCR条带数量以结肠最多,其次是回肠、十二指肠、胃、空肠;抗生素组大鼠胃肠道菌群结构表现出优势菌群的明显降低,并且随着灌胃周期的延长呈现抑制增强的现象。黄连解毒汤高剂量组大鼠胃肠道菌群以大肠杆菌、痤疮丙酸杆菌、沙门氏杆菌等为主要优势菌群,中剂量组大鼠胃肠道菌群以放线菌、绿孢链霉菌、梭状芽孢杆菌、链霉菌、减肥细菌为优势菌群,低剂量组胃肠道优势菌群为痤疮丙酸杆菌。说明黄连解毒汤具有一定的类似抗生素的作用,可抑制胃肠道各段拟杆菌门细菌生长,并有可能促进胃肠道内减肥细菌的增殖。
In this study, the rats were divided into control group, antibiotic group, Huanglian Jiedu Decoction high dose group, Huanglian Jiedu Decoction medium dose group and Huanglianjiedu Decoction low dose group. In each group, 2 mL was administered daily. Distilled gastric water in control group. In the antibiotic group, mice were given ampicillin 56 mg/mL in the first week, ampicillin+streptomycin(56+28) mg/mL in the second week, ampicillin+streptomycin+clindamycin(56+28+50)mg/mL in the third week. Huanglian Jiedu Decoction high, medium and low dose group were given Huanglian Jiedu Decoction 192, 96 and 48 mg/mL respectively. After the administration of the drug, the rats were executed on the 7, 14, 21 days, and the contents of the stomach, duodenum, jejunum, ileum, and colon were collected to extract bacterial DNA. The structural diversity of bacterial flora in each segments of the digestive tract were detected by the repeated consistent sequence polymerase chain reaction(ERIC-PCR) technique. The results show that the Gastrointestinal tract flora structure of control group is relatively stable. ERIC-PCR strips have the largest number in the colon, follow by the ileum, duodenum, stomach, and jejunum. The structure of the digestive tract flora in the antibiotic group shows a significant decrease in the dominant flora, and the inhibition increase with time. The alimentary tract flora of the Huanglian Jiedu Decoction high dose group is mainly dominated by E. coli, Propionibacterium acnes, Salmonella, etc. The medium dose group is mainly dominated by actinomycete Streptomyces, green spore Borrelia, Clostridium spp.,Streptomyces, Akkermansia muciniphila. The low dose group is mainly dominated by Propionibacterium acnes. It indicates that Huanglian Jiedu Decoction has a certain antibiotic like effect which can inhibit the growth of Bacillus phylum bacteria in all sections of the digestive tract, and may promote the proliferation of Akkermansia muciniphila in the digestive tract.
In this study, the rats were divided into control group, antibiotic group, Huanglian Jiedu Decoction high dose group, Huanglian Jiedu Decoction medium dose group and Huanglianjiedu Decoction low dose group. In each group, 2 mL was administered daily. Distilled gastric water in control group. In the antibiotic group, mice were given ampicillin 56 mg/mL in the first week, ampicillin+streptomycin(56+28) mg/mL in the second week, ampicillin+streptomycin+clindamycin(56+28+50)mg/mL in the third week. Huanglian Jiedu Decoction high, medium and low dose group were given Huanglian Jiedu Decoction 192, 96 and 48 mg/mL respectively. After the administration of the drug, the rats were executed on the 7, 14, 21 days, and the contents of the stomach, duodenum, jejunum, ileum, and colon were collected to extract bacterial DNA. The structural diversity of bacterial flora in each segments of the digestive tract were detected by the repeated consistent sequence polymerase chain reaction(ERIC-PCR) technique. The results show that the Gastrointestinal tract flora structure of control group is relatively stable. ERIC-PCR strips have the largest number in the colon, follow by the ileum, duodenum, stomach, and jejunum. The structure of the digestive tract flora in the antibiotic group shows a significant decrease in the dominant flora, and the inhibition increase with time. The alimentary tract flora of the Huanglian Jiedu Decoction high dose group is mainly dominated by E. coli, Propionibacterium acnes, Salmonella, etc. The medium dose group is mainly dominated by actinomycete Streptomyces, green spore Borrelia, Clostridium spp.,Streptomyces, Akkermansia muciniphila. The low dose group is mainly dominated by Propionibacterium acnes. It indicates that Huanglian Jiedu Decoction has a certain antibiotic like effect which can inhibit the growth of Bacillus phylum bacteria in all sections of the digestive tract, and may promote the proliferation of Akkermansia muciniphila in the digestive tract.
引文
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[2] ECKBURG P B,BIK E M,BERNSTEIN C N,et al.Diversity of the human intestinal microbial flora[J].Science,2005,308(5728):1635.
[3] BLAONTON L V,BARRATT M J,CHARBONNEAU M R,et al.Childhood undernutrition,the gut microbiota,and microbiota-directed therapeutics[J].Science,2016,352(6293):1533.
[4] 肖红岩.基于“苦寒伤胃”的抗生素相关性腹泻的发病初探[D].济南:山东中医药大学,2006.
[5] 张文娣.抗生素相关性腹泻模型的构建[D].广州:南方医科大学,2015.
[6] 刘茜明,杨光勇,何光志,等.ERIC-PCR技术分析葛根芩连汤对抗生素相关性腹泻模型消化道菌群结构的影响[J].黑龙江畜牧兽医,2016(06上):23-27.
[7] 赵东凯,王檀.应用黄连解毒汤合苇茎汤治疗肺炎(热毒聚肺型)38例临床观察[J].中国医药指南,2011(9):124-125.
[8] CROW D.Microbiome research in a social world[J].Cell,2018,172(6):1143-1145.
[9] GAO R,ZHU C,LI H,et al.Dysbiosis signatures of gut microbiota along the sequence from healthy,young patients to those with overweight and obesity[J].Obesity,2018,26(2):351-361.
[10] PUEHLER A,KALINOWSKI J.Microbial genomics and metagenomics in human health and disease[J].J Biotechnol,2017,250:1.
[11] PALM N W,DE ZOETE M R,FLAVELL R A.Immune-microbiota interactions in health and disease[J].Clin Immunol,2015,159(2):122-127.
[12] LINDERMANS C A,CALAFIORE M,MERTELSMANN A M,et al.Interleukin-22 promotes intestinal-stem-cell-mediated epithelial regeneration[J].Nature,2015,528(7583):560-564.
[13] SAKAMOTO K,KIM Y G,HARA H,et al.IL-22 controls iron-dependent nutritional immunity against systemic bacterial infections[J].Sci Immunol,2017,2(8):8371.
[14] YAN L,WANG J,XU D,et al.Comparative study of single/combination use of Huang-Lian-Jie-Du decoction and berberine on their protection on sepsis induced acute liver injury by NMR metabolic profiling[J].J Pharm Biomed Anal,2017,145:794-804.
[15] XU Y,GUO S,CHEN G,et al.Evaluation of anti-sepsis activity by compounds with high affinity to lipid a from HuanglianJiedu decoction[J].Immunopharmacol Immunotoxicol,2017,39(6):364-370.
[16] 刘磊,姜鹏,窦圣姗,等.黄连解毒汤的化学及药理学研究进展[J].中草药,2008,39(6):935-938.
[17] 罗海华.黄连解毒汤对小鼠消化道菌群的影响[D].广州:广州中医药大学,2008.
[18] GOPALAKRISHNAN V,SPENCER C N,NEZI L,et al.Gut microbiome modulates response to anti-PD-1 immunotherapy in melanoma patients[J].Science,2018,359(6371):97-103.