Bovine serum albumin as a resuscitation promoting factor for viable but non-culturable <i>Mycobacterium tuberculosis</i> via the activation of protein kinase A-dependent cellular processes
Bovine serum albumin as a resuscitation promoting factor for viable but non-culturable <i>Mycobacterium tuberculosis</i> via the activation of protein kinase A-dependent cellular processes
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牛血清白蛋白通过激活蛋白激酶 A 依赖性细胞过程作为活的但不可培养的结核分枝杆菌的复苏促进因子
DOI：
10.1101/2021.11.22.468319
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发表时间：
2021
期刊：
影响因子：
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通讯作者：
Mitarai Satoshi
中科院分区：
文献类型：
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作者：
Morishige Yuta;Murase Yoshiro;Chikamatsu Kinuyo;Aono Akio;Igarashi Yuriko;Shimomura Yoshiko;Hosoya Makiko;Kamada Keisuke;Yamada Hiroyuki;Takaki Akiko;Mitarai Satoshi
结核分枝杆菌（Mtb）H37 Ra菌株在用NADH氧化酶抑制剂（二苯基碘鎓 [DPI]）处理后迅速进入活的但不可培养（VBNC）状态，并被胎牛血清（FBS）重新激活。然而，FBS诱导再激活的机制尚不清楚。我们试图用M.方法首先，我们评估DPI对H37 Rv的可培养性、活力和耐酸性变化的影响。其次，我们检测了人血清白蛋白、蛋清白蛋白和抗氧化剂对DPI诱导的VBNC细胞的再激活促进作用。结果DPI处理后，H37 Rv细胞处于VBNC状态，活细胞比例高，可培养细胞比例低，细胞耐酸性下降，脂质积累减少。FBS和BSA均可使H37Rv重激活。与我们的预期相反，只有人血清白蛋白具有与BSA相似的恢复效果。腺苷酸环化酶抑制SQ22536没有显着的效果上的再激活，但是，抑制分枝杆菌蛋白激酶H89和staurosporine强烈抑制BSA诱导reactivation.ConclusionDPI诱导的VBNC Mtb细胞可能会通过激活蛋白激酶依赖的细胞分裂过程中通过与BSA的相互作用被重新激活。
ObjectiveMycobacterium tuberculosis (Mtb) H37Ra strain has been reported to rapidly enter the viable but non-culturable (VBNC) state following treatment with an NADH oxidase inhibitor (diphenyleneiodonium [DPI]) and to be reactivated by fetal bovine serum (FBS). However, the mechanism underlying FBS-induced reactivation is unclear. We tried to reveal the mechanism of FBS-induced reactivation using M. tuberculosis H37Rv.MethodsFirst, we evaluated the effect of DPI on culturability, viability and changes of acid-fastness toward H37Rv. Secondly, we measured the reactivation-promoting effects of human serum albumin, egg-white albumin and antioxidative agents in DPI-induced VBNC cells. We also inhibited adenylyl cyclase and protein kinase which is the downstream of adenylyl cyclase to evaluate the influence to reactivation capacity of bovine serum albumin (BSA).ResultsDPI treatment induced VBNC state in H37Rv, resulting in a high proportion of viable cells but a low proportion of culturable cells, loss of acid-fastness and lipid-accumulation. Not only FBS but also BSA alone could reactivate H37Rv. Contrary to our expectation, only human serum albumin had a similar restorative effect to BSA. The inhibition of adenylyl cyclase by SQ22536 did not have a significant effect on reactivation; however, the inhibition of mycobacterial protein kinase by H89 and staurosporine strongly suppressed the BSA-induced reactivation.ConclusionDPI-induced VBNC Mtb cells may be reactivated via the activation of protein kinase-dependent cell division processes through interaction with BSA.