异烟肼影响重组耻垢分支杆菌Rv1776c基因表达的实验研究

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。

The influence of isoniazid on the expression ofrecombinant Mycobacterium smegmatis Rv1776c gene

Objective To study the influence of classical anti-tuberculosis drug isoniazid on the growth and proliferation of recombinant Mycobacterium smegmatis and the expression of recombinant Mycobacterium smegmatis Rv1776c gene. Methods The recombinant strain MS-Rv1776c was inoculated into LB medium as control group. The experimental group was treated with isoniazid, and the A600 value was measured at different time points. The proliferation curve was plotted according to the measured value of A. The bacterial DNA was extracted to detect the influence of isoniazid on the expression of Rv1776c gene by RT-PCR method. The influence of isoniazid on the expression of Rv1776c protein was detected by immunoblotting with SDS-PAGE and Western blot. Results Isoniazid had no significant influence on the proliferation of recombinant Mycobacterium smegmatis in both groups (P>0.05). The expression of Rv1776c gene was inhibited by isoniazid (P<0.05). SDS-PAGE and Western blot analysis showed that isoniazid significantly reduced the expression of ERv1776c protein (P<0.05). Conclusion Isoniazid had no influence on the proliferation of recombinant Mycobacterium smegmatis, but inhibited the expression of Rv1776c gene and the protein in Mycobacterium tuberculosis. The results provided experimental evidence for the study on the prevention of Mycobacterium tuberculosis from dormant to early recovery and active period.