变性梯度凝胶电泳法分析黄柏对大鼠胃肠道菌群的影响
|
摘要
目的:应用DGGE技术分析黄柏对大鼠胃、十二指肠、空肠、回肠、结肠部位菌群数量的影响,为探索黄柏的肠菌群调节作用积累数据。方法:饲养大鼠共72只,黄柏水煎液分高、中、低剂量进行灌胃,其中空白对照组18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),提取细菌基因组DNA,应用DGGE技术分析各样本不同给药时间下菌群数量的变化。结果:在黄柏作用的影响下,胃部低剂量组一周,中剂量组三周及高剂量组二周三周与空白组差异较明显(P<0.05);十二指肠低剂量组一周,低、中、高剂量组第三周较空白组有明显差异(P<0.05);空肠部位低、中、高剂量组第三周较空白组有明显差异(P<0.05);回肠部位低剂量组二、三周,中剂量组一、二、三周及高剂量组一周较空白组有明显差异(P<0.05);结肠部位中剂量组一、二、三周较空白组有明显差异(P<0.05)。结论:黄柏长期给药对大鼠十二指肠部位菌群多样性起抑制作用,对胃、空肠、回肠及结肠部位的菌群多样性起到一定促进作用。
Objective:To accumulate data for investigating the regulation of intestinal microflora of Phellodendron chinense by using DGGE technology to analyse the effect of cortex phellodendri on the intestinal flora in rat's stomach,duodenum,jejunum,ileum and colon. Methods: 72 rats were raised and gavaged with decoction of cortex phellodendri with high,medium and low doses. Among them,18 rats were in the blank control group. Gathered rat stomach,duodenum,jejunum,ileum and colon( including content) in the first,second and third week respectively and extracted bacterial genome DNA. Then analyzed the change of the number of flora in different samples at different dosing time by DGGE Technology. Results: Under the influence of cortex Phellodendron,compared with the blank group of the stomach,the differences were obvious between one week of the low-dose group,the three weeks of the medium-dose group and the two and three weeks of high-dose group( P < 0. 05). Compared with the blank group,in the group of duodemun,there was a significant difference between the low-dose,medium-doe and high-dose groups in the third week( P < 0. 05). There was significant difference in the third week of the low,medium and high dose group of jejunum compared with that of the blank group( P < 0. 05). In ileum,there were significant differences in low-dose group in two,three weeks,medium-dose group in one,two three weeks and in high-dose group in one week compared with the bland group( P < 0. 05). In colon,there were obvious differences in medium-dose group in one,two and three weeks compared with the blank group( P < 0. 05). Conclusion: Long-term administration of Cortex Phellodendri could inhibite the diversity of duodenal flora in rats,and it played a certain role in promoting the flora diversity in the stomach,jejunum,ileum and colon.
Objective:To accumulate data for investigating the regulation of intestinal microflora of Phellodendron chinense by using DGGE technology to analyse the effect of cortex phellodendri on the intestinal flora in rat's stomach,duodenum,jejunum,ileum and colon. Methods: 72 rats were raised and gavaged with decoction of cortex phellodendri with high,medium and low doses. Among them,18 rats were in the blank control group. Gathered rat stomach,duodenum,jejunum,ileum and colon( including content) in the first,second and third week respectively and extracted bacterial genome DNA. Then analyzed the change of the number of flora in different samples at different dosing time by DGGE Technology. Results: Under the influence of cortex Phellodendron,compared with the blank group of the stomach,the differences were obvious between one week of the low-dose group,the three weeks of the medium-dose group and the two and three weeks of high-dose group( P < 0. 05). Compared with the blank group,in the group of duodemun,there was a significant difference between the low-dose,medium-doe and high-dose groups in the third week( P < 0. 05). There was significant difference in the third week of the low,medium and high dose group of jejunum compared with that of the blank group( P < 0. 05). In ileum,there were significant differences in low-dose group in two,three weeks,medium-dose group in one,two three weeks and in high-dose group in one week compared with the bland group( P < 0. 05). In colon,there were obvious differences in medium-dose group in one,two and three weeks compared with the blank group( P < 0. 05). Conclusion: Long-term administration of Cortex Phellodendri could inhibite the diversity of duodenal flora in rats,and it played a certain role in promoting the flora diversity in the stomach,jejunum,ileum and colon.
引文
[1]陈建章,尹建康,侯春久,等.黄连黄柏对脾胃虚寒实验大鼠模型的影响[J].中国中医药现代远程教育,2015,13(10):134-136.
[2]徐珊,张凡,刘蓬蓬,等.基于大鼠物质、能量代谢研究炮制对黄柏药性的影响[J].中药材,2015,38(09):1835-1841.
[3]陈传蓉,李必刚,张林,等.黄柏历代临床用量分析[J].中医杂志,2014,55(13):1142-1145.
[4]Ferguson A S,Huang W E,Lawson K A,et al.Microbial analysis of soil and groundwater from a gasworks site and comparison with asequenced biological reactive barrier remediation process[J].J.Appl Microbiol.2007,102(5):1227~1238.
[5]吴云,段智璇,陈瑞,等.PCR-DGGE法检测大鼠胃肠道菌群结构的电泳条件探讨[J].贵阳中医学院学报,2016,38(03):19-22.
[6]周悦芳,范培红.中药免疫调节作用研究进展[J].时珍国医国药,2017,28(01):204-207.
[7]邱琳.中药对胃肠道动力调节作用的研究概况[J].中医临床研究,2015,35(07):147-148.
[8]郑昱婧,宋淑亮,梁浩,等.中药多糖在抗肿瘤免疫中的研究进展[J].生命的化学,2016,36(05):601-606.
[9]李丹丹,江培,杨书美,等.黄柏的化学成分、药理作用及临床应用研究进展[J].黑龙江医药,2014,27(03):601-605.
[10]任锡禄,郝高庭,窦志芳.黄柏治疗慢性骨髓炎临床应用及药理研究进展[J].山西中医学院学报,2016,17(01):63-64.
[11]段智璇,张柏岳,瞿慧琴,等.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[J].贵阳中医学院学报,2016,38(06):1-8+10.
[12]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.
[2]徐珊,张凡,刘蓬蓬,等.基于大鼠物质、能量代谢研究炮制对黄柏药性的影响[J].中药材,2015,38(09):1835-1841.
[3]陈传蓉,李必刚,张林,等.黄柏历代临床用量分析[J].中医杂志,2014,55(13):1142-1145.
[4]Ferguson A S,Huang W E,Lawson K A,et al.Microbial analysis of soil and groundwater from a gasworks site and comparison with asequenced biological reactive barrier remediation process[J].J.Appl Microbiol.2007,102(5):1227~1238.
[5]吴云,段智璇,陈瑞,等.PCR-DGGE法检测大鼠胃肠道菌群结构的电泳条件探讨[J].贵阳中医学院学报,2016,38(03):19-22.
[6]周悦芳,范培红.中药免疫调节作用研究进展[J].时珍国医国药,2017,28(01):204-207.
[7]邱琳.中药对胃肠道动力调节作用的研究概况[J].中医临床研究,2015,35(07):147-148.
[8]郑昱婧,宋淑亮,梁浩,等.中药多糖在抗肿瘤免疫中的研究进展[J].生命的化学,2016,36(05):601-606.
[9]李丹丹,江培,杨书美,等.黄柏的化学成分、药理作用及临床应用研究进展[J].黑龙江医药,2014,27(03):601-605.
[10]任锡禄,郝高庭,窦志芳.黄柏治疗慢性骨髓炎临床应用及药理研究进展[J].山西中医学院学报,2016,17(01):63-64.
[11]段智璇,张柏岳,瞿慧琴,等.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[J].贵阳中医学院学报,2016,38(06):1-8+10.
[12]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.