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目的研究经典抗结核药物异烟肼对重组耻垢分支杆菌生长增殖及其Rv1776c基因表达的影响。方法将重组菌MS-Rv1776c接种于LB培养基培养作为对照组,实验组给予异烟肼药物处理,不同时间点取菌液测量A600值,根据所测A值绘制增殖曲线;提取菌液DNA以RT-PCR方法检测异烟肼对Rv1776c基因表达的影响;免疫印迹法SDS-PAGE及Western blot检测异烟肼对Rv1776c蛋白表达的影响。结果异烟肼对两组重组耻垢分枝杆菌增殖无显著影响(P>0.05);异烟肼可抑制Rv1776c基因的表达(P<0.05);SDS-PAGE及Western blot检测发现异烟肼可显著降低ERv1776c蛋白的表达(P<0.05)。结论异烟肼对重组耻垢分支杆菌的增殖无影响,但可抑制结核分枝杆菌Rv1776c基因及其表达的蛋白。该结果对研究结核菌从休眠菌到复苏初期及活跃期的药物预防提供了实验依据。
Objective To study the effects of isoniazid, a classical anti-tuberculosis drug, on the growth and proliferation of recombinant Mycobacterium smegmatis and the expression of Rv1776c. Methods The recombinant strain MS-Rv1776c was inoculated into LB culture medium as the control group. The experimental group was treated with isoniazid. The A600 value was measured at different time points and the proliferation curve was drawn according to the measured value of A. The bacterial DNA was extracted The effect of isoniazid on Rv1776c gene expression was detected by RT-PCR. The expression of Rv1776c protein was detected by SDS-PAGE and Western blot. Results Isoniazid had no significant effect on the proliferation of recombinant Mycobacterium smegmatis (P> 0.05). Isoniazid inhibited the expression of Rv1776c (P <0.05). The results of SDS-PAGE and Western blot showed that isoniazid Significantly decreased the expression of ERv1776c protein (P <0.05). Conclusion Isoniazid has no effect on the proliferation of recombinant Mycobacterium smegmatis, but inhibits the M. tuberculosis Rv1776c gene and its expressed protein. The results provide an experimental basis for the study on the drug prevention of Mycobacterium tuberculosis from dormant bacteria to early and active resuscitation.