实时荧光定量PCR法分析苦参对大鼠胃肠道大肠杆菌数量的影响
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摘要
目的:应用实时荧光定量PCR技术分析口服苦参对大鼠胃、十二指肠、空肠、回肠、结肠部位以及粪便内大肠杆菌数量的影响,探索苦参对胃肠道中大肠杆菌菌群调节作用。方法:常规饲养大鼠150只,苦参水煎液分高、中、低剂量灌胃,另设空白对照组与抗生素组各10只。分别在第一、二、三周时采集大鼠胃、十二指肠、空肠、回肠、结肠各段(含内容物)以及粪便,提取细菌基因组DNA,采用实时荧光定量PCR法(绝对定量方法)检测各样本不同给药时间点大肠杆菌的数量。结果:(1)与空白对照组相比较:各给药组十二指肠部位在第一、二、三周时,大肠杆菌数量显著降低(P<0.01);在不同剂量组不同给药时间点,胃部、空肠、回肠部位以及粪便中大肠杆菌数量均有不同程度降低(P<0.05)。(2)与抗生素组比较:低、中、高剂量组给药二周后,十二指肠、空肠、回肠部位大肠杆菌数量明显偏低(P<0.05),而结肠及粪便部位在给药一周后高剂量组大肠杆菌数量明显偏高(P<0.01)。结论:苦参水煎液灌胃可造成大鼠十二指肠、空肠、回肠部位大肠杆菌菌群数量降低,其中对十二指肠部位大肠杆菌数量的影响随时间推移呈递减趋势,与抗生素的作用基本一致。
Objective: To analyze the effect of Sophora flavescens Alt. on the number of Escherichia coli.in the stomach,duodenum,jejunum,ileum,colon and excrement of SD rats by the method of real-time fluorescent quantitative PCR and study the regulating effect of Sophora flavescens Alt. on escherichia coli bacteria in gastrointestinal tract. Methods: Fed 150 SD rats with conventional way,and gave decoction of Sophora flavescens Alt. by gavage with high,medium and low dose. Meanwhile,a blank control group and an antibiotic group were set up with 10 rats in each group. SD rats' stomach,duodenum,jejunum,ileum parts and colon paragraphs( including their contents) and feces were gathered in the first,second and third weeks,and bacterial genome DNA was extracted. Detected the number of Escherichia coli at different time points of administration by using real-time fluorescent quantitative PCR method. Results:( 1) Compared with the blank control group: in the dosage group,the number of Escherichia coli in the duodenal parts decreased significantly at the first,second and third weeks,( P < 0. 01); In the different dose group,at different administration time,the number of Escherichia coli. in the stomach,jejunum,ileum parts,and feces reduced to some extent( P < 0. 05).( 2)( 2) Compared with the antibiotics group: two weeks after administration,the number of Escherichia coli. in duodenum,jejunum and ileum in low,medium and high dose groups reduced significantly( P < 0. 05),while in the high dose group,the number of Escherichia coli. in colon parts and feces increased obviously after one week of drug delivery( P < 0. 01). Conclusion: Intragastric administration with Sophora flavescens Alt. decoction can reduce the number of Escherichia coli bacteria in rats' duodenum,jejunum,ileum parts. Its impact on the number of Escherichia coli in duodenal parts decreased gradually over time,which is consistent with the effect of antibiotics.
Objective: To analyze the effect of Sophora flavescens Alt. on the number of Escherichia coli.in the stomach,duodenum,jejunum,ileum,colon and excrement of SD rats by the method of real-time fluorescent quantitative PCR and study the regulating effect of Sophora flavescens Alt. on escherichia coli bacteria in gastrointestinal tract. Methods: Fed 150 SD rats with conventional way,and gave decoction of Sophora flavescens Alt. by gavage with high,medium and low dose. Meanwhile,a blank control group and an antibiotic group were set up with 10 rats in each group. SD rats' stomach,duodenum,jejunum,ileum parts and colon paragraphs( including their contents) and feces were gathered in the first,second and third weeks,and bacterial genome DNA was extracted. Detected the number of Escherichia coli at different time points of administration by using real-time fluorescent quantitative PCR method. Results:( 1) Compared with the blank control group: in the dosage group,the number of Escherichia coli in the duodenal parts decreased significantly at the first,second and third weeks,( P < 0. 01); In the different dose group,at different administration time,the number of Escherichia coli. in the stomach,jejunum,ileum parts,and feces reduced to some extent( P < 0. 05).( 2)( 2) Compared with the antibiotics group: two weeks after administration,the number of Escherichia coli. in duodenum,jejunum and ileum in low,medium and high dose groups reduced significantly( P < 0. 05),while in the high dose group,the number of Escherichia coli. in colon parts and feces increased obviously after one week of drug delivery( P < 0. 01). Conclusion: Intragastric administration with Sophora flavescens Alt. decoction can reduce the number of Escherichia coli bacteria in rats' duodenum,jejunum,ileum parts. Its impact on the number of Escherichia coli in duodenal parts decreased gradually over time,which is consistent with the effect of antibiotics.
引文
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[7]刘崇海,杨锡强,刘恩梅,等.抗生素诱导肠道菌群失调在RSV感染小鼠模型上对肺部Th1/Th2的影响[J].现代预防医学,2011,38(12):2374-2377.
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[2]许柳青,秦林,王加锋,等.苦寒中药对胃肠功能的影响研究概况[J].山东中医药大学学报,2010,34(6):554-555.
[3]文细毛,任南,徐秀华,等.全国医院感染监控网医院感染病原菌分布及耐药性分析[J].中华医院感染学杂志,2002,04:4-7.
[4]黄金莲,朱彦仁,应福余,等.主要医院感染病原菌的变迁及其耐药性分析[J].中国微生态学杂志,2005,17(4):290-292.
[5]Mizushima T,Sasaki M,Ando T,et al.Blockage of angiotensinⅡtype 1receptor regulates TNF-alpha-induced MAd CAM-1 expression via inhibitionof NFkappa B translocation to the nucleus and ameliorates colitis[J].Am J Physiol Gastrointest Liver Physiol,2010,298(2):G255-266.
[6]贺洁,彭学博.清热药保留灌肠配合西药治疗溃疡性结肠炎48例[J].陕西中医,2013,34(1):24-25.
[7]刘崇海,杨锡强,刘恩梅,等.抗生素诱导肠道菌群失调在RSV感染小鼠模型上对肺部Th1/Th2的影响[J].现代预防医学,2011,38(12):2374-2377.
[8]邱慧卿,徐建光.苦参素注射液联合中药灌肠治疗溃疡性结肠炎临床观察[J].新中医,2015,11:69-71.
[9]岳文鹏,田维毅.肠菌生物转化对白头翁水提液体外抑菌作用的影响[J].贵阳中医学院学报,2013,02:47-49.