对非复制期结核分枝杆菌活性测定模型的建立及应用

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中图分类号：R978.1 文献标志码：A

Abstract Objective Two non-replicating phase models of Mycobacterium tuberculosis in hypoxic and nutrient starvation cultures were developed for drug activity assays in vitro to provide asistance inthe development of drugs that were efective against non-replicating Mycobacterium tuberculosis with the aim of shortening the course of therapy.MethodsMycobacterium tuberculosis was induced into the non-replicative phase by gradually decreasing the oxygen concentration and nutritional starvation.The gene expression changes of Mycobacterium tuberculosis entering into the non-replicative state were analyzed by using transcriptome sequencing，and the two models were applied to evaluate the activities of 5-NPand different anti-tuberculosis drugs.ResultsThe models of non-replicating Mycobacterium tuberculosis induced by hypoxia and nutrient starvation were successfully established.The transcriptome data revealed 1，636 and1，814 diffrential genes in the two models，respectively.Both models caused changes in oxidative phosphorylationand metabolic pathways.In addition，genes related to the arabinogalactan synthesis pathway showed significant changes in the hypoxia model，while diferentially expressed genes were enriched in the carbon metabolism and amino acid synthesis pathways in the starvation model. Some drugs had significantly different activities against Mycobacterium tuberculosis in the two models.The 5μg/mL concentration group of 5-NP in the starvation model reduced the log10CFU by 2.72，better than that of rifampicin in the high-concentration group （ΔLog10CFU20μg/mL=2.55） . The 20μg/mL concentration groupof 5-NP in the hypoxia model reduced the Log10CFU by 1.05， which was comparable to thatofpretomanid （ΔLog10CFU20μg/mL=1.32） . Metronidazole exhibitedsignificantactivityonlyagainst Mycobacteriumtubereulosis inthe hypoxicmodel （ΔLog10CFU5μg/mL=2.00） ConclusionInthis study，two models for investigating theactivityof non-replicating Mycobacterium tuberculosis induced by hypoxia and starvation were successfully developed and applied，providing asistance for the research and development of anti-tuberculosis drugs， especially for those possessing efficacy against non-replicating Mycobacterium tuberculosis.

KeyWordsMycobacterium tuberculosis;Non-replication;Hyoxia; Stavation; Transcriptome

结核病是由结核分枝杆菌（Mycobacteriumtuberculosis，MTB）引起的一种严重危害人类健康的慢性传染性疾病。（剩余12858字）