黄芩苷和沉默基因acrA、marA和acrD对耐药大肠埃希菌环丙沙星敏感性的影响

发布时间：2018-05-31 10:31

  本文选题：反义寡核苷酸 + 大肠埃希菌　； 参考：《中国兽医学报》2017年02期

【摘要】：为探讨反义寡核苷酸(ASODN)沉默acrA、marA和acrD基因前后,黄芩苷对临床耐药大肠埃希菌(E.coli)环丙沙星敏感性的影响,设计并合成3个基因的ASODN引物,设置空白对照组、黄芩苷组、ASODN沉默组、ASODN沉默+黄芩苷组,分别转染到临床耐药菌感受态细胞中,涂布于一定浓度环丙沙星平板上,培养观察菌落生长情况并计数。结果发现,相对于空白对照组,黄芩苷组菌落数显著降低;acrA、acrD和marA基因沉默组与空白对照组相比,除了acrD基因沉默1号菌株和marA基因沉默的Y17菌株外,其余菌株在环丙沙星平板上的菌落数也出现不同程度地下降,具有显著性差异;acrA、acrD和marA基因沉默+黄芩苷组与黄芩苷组相比,多数菌株在环丙沙星平板上的菌落数均出现不同程度地下降;acrA基因沉默+黄芩苷组与acrA基因沉默组相比,4株菌株变化差异不显著,Y17号菌株菌落数有所上升,J45号菌株在该基因沉默后,环丙沙星平板上的菌落数出现下降的趋势,差异显著;acrD基因沉默+黄芩苷组与acrD基因沉默组相比,除了1号菌株变化差异不显著外,其余5株菌株变化差异显著,但Y17号菌株菌落数上升;marA基因沉默+黄芩苷组与marA基因沉默组相比,除了Y17、Y35这2株菌株变化差异显著外,其余4株菌株变化差异不显著。以上结果表明,黄芩苷主要通过抑制acrA和marA发挥外排抑制作用,acrD不是其增强环丙沙星敏感性的主要靶点。
[Abstract]:In order to investigate the effect of baicalin on the susceptibility of clinical resistant Escherichia coli to ciprofloxacin before and after antisense oligonucleotide silencing acrAlomara and acrD genes, ASODN primers of three genes were designed and synthesized, and blank control group was set up. Baicalin group was silenced by ASODN silencing baicalin group and transfected into the competent cells of clinical drug-resistant bacteria and coated on a certain concentration of ciprofloxacin plate to observe the colony growth and count. The results showed that the colony number of baicalin group was significantly lower than that of blank control group, except acrD gene silencing strain 1 and marA gene silencing strain Y17. The colony number of other strains on ciprofloxacin plate also decreased to some extent, and there was significant difference between baicalin group and baicalin group in acrAgnacrD and marA gene silencing. The colony number of most strains on ciprofloxacin plate was decreased in varying degrees. There was no significant difference between acrA gene silencing baicalin group and acrA gene silencing group. The colony number of Y17 strain increased to J45. After the gene was silenced, The number of colonies on ciprofloxacin plate decreased. The difference was significant between baicalin silencing group and acrD gene silencing group, except for strain 1, there were significant differences among the other 5 strains. However, there was no significant difference between Y17 and marA gene silencing baicalin group, except Y17 Y35 strain, except Y17 Y35 gene silencing baicalin group and Y17 Y35 gene silencing group, but there was no significant difference in the other 4 strains in comparison with the marA gene silencing group. These results indicate that baicalin exerts efflux inhibition mainly by inhibiting acrA and marA. AcrD is not the main target for enhancing ciprofloxacin sensitivity.
【作者单位】： 河南农业大学牧医工程学院;
【基金】：国家自然科学基金资助项目(U1304328) 河南省科技厅资助项目(132300410113)
【分类号】：R285
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本文编号：1959306