State Key Laboratory of Agriculture Microbiology, Huazhong Agricultural University, Wuhan, Hubei, 430070, PR China; College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei, 430070, PR China.
College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei, 430070, PR China.
Microb Pathog. 2017 Nov;112:243-253. doi: 10.1016/j.micpath.2017.09.055. Epub 2017 Sep 28.
Resistance to anti-tuberculosis drugs is a formidable obstacle to effective tuberculosis (TB) treatment and prevention globally. New forms of multidrug, extensive drug and total drug resistance Mycobacterium tuberculosis (Mtb) causing a serious threat to human as well as animal's population. Mtb shows diverse adaptability under stress conditions especially antibiotic treatment, however underlying physiological mechanism remained elusive. In present study, we investigated Mtb's response and adaptation with reference to gene expression during sub-lethal kanamycin exposure. Mtb were cultured under sub-lethal drug and control conditions, where half were sub-cultured every 3-days to observe serial adaptation under same conditions and the remaining were subjected to RNA-seq. We identified 98 up-regulated and 198 down-regulated responsive genes compared to control through differential analysis, of which Ra1750 and Ra3160 were the most responsive genes. In adaptive analysis, we found Ra1750, Ra3160, Ra3161, Ra3893 and Ra2492 up-regulation at early stage and gradually showed low expression levels at the later stages of drug exposure. The adaptive expression of Ra1750, Ra3160 and Ra3161 were further confirmed by real time qPCR. These results suggested that these genes contributed in Mtb's physiological adaptation during sub-lethal kanamycin exposure. Our findings may aid to edify these potential targets for drug development against drug resistance tuberculosis.
抗结核药物耐药性是全球有效治疗和预防结核病的一个巨大障碍。新型多药、广泛耐药和全耐药结核分枝杆菌（Mtb）对人类和动物的种群构成了严重威胁。Mtb 在压力条件下，尤其是在抗生素治疗下，表现出多样的适应性，但潜在的生理机制仍不清楚。在本研究中，我们研究了 Mtb 在亚致死性卡那霉素暴露时的基因表达，以探讨其对亚致死性卡那霉素暴露的反应和适应。将 Mtb 在亚致死性药物和对照条件下培养，其中一半每 3 天进行亚培养，以观察相同条件下的连续适应，其余的进行 RNA-seq 分析。通过差异分析，我们发现与对照相比，有 98 个上调和 198 个下调的响应基因，其中 Ra1750 和 Ra3160 是最敏感的基因。在适应性分析中，我们发现 Ra1750、Ra3160、Ra3161、Ra3893 和 Ra2492 在早期上调，在药物暴露的后期逐渐表现出低表达水平。Ra1750、Ra3160 和 Ra3161 的适应性表达通过实时 qPCR 进一步得到证实。这些结果表明，这些基因在 Mtb 亚致死性卡那霉素暴露时的生理适应中发挥了作用。我们的发现可能有助于修饰这些潜在的药物开发靶点，以对抗耐药性结核病。
