小毛莨内酯对重组耻垢分枝杆菌eis基因表达的影响

Influence of ternatolide on the expression of eis gene in recombinant Mycobacterium smegmatis

目的研究小毛莨内酯(Ternatolide,Tern)对重组耻垢分枝杆菌生长增殖及其eis基因表达的影响。方法将5×105MS-eis-PMV261接种于LB培养基培养并加入200 mg/L的Tern作为实验组,MS-eis-PMV261单独培养作为对照,不同时间点测量取菌液波长为600 nm的A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Tern对eis基因表达的影响;免疫印迹法SDS-PAGE及Western Blot检测Tern对EIS蛋白表达的影响。结果 Tern对两组重组耻垢分枝杆菌增殖差异无统计学意义(P>0.05);Tern可抑制eis基因的表达(P<0.05);SDS-PAGE及Western Blot检测发现Tern可显著降低EIS蛋白的表达(P<0.05)。结论 Tern对重组耻垢分枝杆菌的增殖无影响,但可抑制结核分枝杆菌eis基因及其表达的蛋白。该结果对研究持留性结核分枝杆菌的药物治疗提供了实验依据。

Objective To explore the influence of Ternatolide on the growth of recombinant Mycobacterium smegmatis,and the expression of eis gene and protein.Methods 5×105 of MS-eis-PMV261 was inoculated in LB medium as the control group.MS-eis-PMV261 was cultured alone as the experiment group which was treated with 200 mg/L of Ternatolide.Inocula were collected at different time point and then the A values at 600 nm wavelength were measured to get the proliferation curve.The eis gene and protein expression in Mycobacterium smegmatis treated as above was detected by RT-PCR and Immunoblot analyses respectively.Results Ternatolide showed no influence on the growth of Mycobacterium smegmatis（P0.05）,but inhibited the expression of eis gene and protein which played a great role in the persistency of Mycobacterium tuberculosis（P0.05）.SDS-PAGE and Western blot showed that Ternatolide further decreased the expression of EIS protein（P0.05）.Conclusion Ternatolide has no effect on the growth of recombinant Mycobacterium smegmatis,however,it can inhibit the expression of eis gene and protein of Mycobacterium tuberculosis.The results of the study can provide experimental basis for the treatment of persistant Mycobacterium tuberculosis.