低浓度顺铂通过磷酸化 p53 相关凋亡通路抑制巨噬细胞中的分枝杆菌增殖。
Inhibition of mycobacteria proliferation in macrophages by low cisplatin concentration through phosphorylated p53-related apoptosis pathway.
机构信息
Department of Pathogenic Biology, College of Basic Medicine, Chongqing Medical University, Chongqing, China.
Hospital-Acquired Infection Control Department, First People's Hospital of Jintang County, Chengdu, China.
出版信息
PLoS One. 2023 Jan 31;18(1):e0281170. doi: 10.1371/journal.pone.0281170. eCollection 2023.
BACKGROUND
Drug resistance is a prominent problem in the treatment of tuberculosis, so it is urgent to develop new anti- tuberculosis drugs. Here, we investigated the effects and mechanisms of cisplatin (DDP) on intracellular Mycobacterium smegmatis to tap the therapeutic potential of DDP in mycobacterial infection.
RESULTS
Macrophages infected with Mycobacterium smegmatis were treated with DDP alone or combined with isoniazid or rifampicin. The results showed that the bacterial count in macrophages decreased significantly after DDP (≤ 6 μg/mL) treatment. When isoniazid or rifampicin was combined with DDP, the number of intracellular mycobacteria was also significantly lower than that of isoniazid or rifampicin alone. Apoptosis of infected cells increased after 24 h of DDP treatment, as shown by flow cytometry and transmission electron microscopy detection. Transcriptome sequencing showed that there were 1161 upregulated and 645 downregulated differentially expressed genes (DEGs) between the control group and DDP treatment group. A Trp53-centered protein interaction network was found based on the top 100 significant DEGs through STRING and Cytoscape software. The expression of phosphorylated p53, Bax, JAK, p38 MAPK and PI3K increased after DDP treatment, as shown by Western blot analysis. Inhibitors of JAK, PI3K or p38 MAPK inhibited the increase in cell apoptosis and the reduction in the intracellular bacterial count induced by DDP. The p53 promoter Kevetrin hydrochloride scavenges intracellular mycobacteria. If combined with DDP, Kevetrin hydrochloride could increase the effect of DDP on the elimination of intracellular mycobacteria. In conclusion, DDP at low concentrations could activate the JAK, p38 MAPK and PI3K pathways in infected macrophages, promote the phosphorylation of p53 protein, and increase the ratio of Bax to Bcl-2, leading to cell apoptosis, thus eliminating intracellular bacteria and reducing the spread of mycobacteria.
CONCLUSION
DDP may be a new host-directed therapy for tuberculosis treatment, as well as the p53 promoter Kevetrin hydrochloride.
背景
耐药性是结核病治疗中的一个突出问题，因此迫切需要开发新的抗结核药物。在这里，我们研究了顺铂（DDP）对细胞内耻垢分枝杆菌的影响及其机制，以挖掘 DDP 在分枝杆菌感染中的治疗潜力。
结果
用 DDP 单独或联合异烟肼或利福平处理感染耻垢分枝杆菌的巨噬细胞。结果表明，DDP（≤6μg/ml）处理后巨噬细胞内细菌数量明显减少。当异烟肼或利福平与 DDP 联合使用时，细胞内分枝杆菌的数量也明显低于异烟肼或利福平单独使用时。用 DDP 处理 24 小时后，通过流式细胞术和透射电镜检测，发现感染细胞的凋亡增加。转录组测序显示，对照组和 DDP 处理组之间有 1161 个上调和 645 个下调的差异表达基因（DEGs）。通过 STRING 和 Cytoscape 软件，基于前 100 个显著 DEGs 构建了一个色氨酸 53 中心蛋白相互作用网络。用 Western blot 分析发现，DDP 处理后磷酸化 p53、Bax、JAK、p38 MAPK 和 PI3K 的表达增加。JAK、PI3K 或 p38 MAPK 的抑制剂抑制了 DDP 诱导的细胞凋亡增加和细胞内细菌数量减少。p53 启动子 Kevetrin 盐酸盐可清除细胞内分枝杆菌。如果与 DDP 联合使用，Kevetrin 盐酸盐可以增强 DDP 对消除细胞内分枝杆菌的作用。综上所述，低浓度 DDP 可激活感染巨噬细胞中的 JAK、p38 MAPK 和 PI3K 通路，促进 p53 蛋白磷酸化，增加 Bax 与 Bcl-2 的比值，导致细胞凋亡，从而清除细胞内细菌，减少分枝杆菌的传播。
结论
DDP 可能是一种新的抗结核宿主定向治疗药物，以及 p53 启动子 Kevetrin 盐酸盐。
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