目的:使用大麻,含有多种抗菌药物,可能是牙周炎的危险因素。我们假设多种口服螺旋体对植物大麻素具有抗性,并且大麻二酚(CBD)将充当环境应激源,而Denticola密螺旋体将在转录上做出反应,从而提供对螺旋体生存策略的初步见解。
方法:在存在和不存在生理相关的植物大麻素剂量的情况下,用分光光度法监测口腔螺旋体的生长。通过RNAseq确定的对植物大麻素暴露的转录反应,在已鉴定的完全测序的口腔螺旋体中,使用qRT-PCR和直系同源物验证了特定的基因活性通量。
结果:多种口腔螺旋体菌株对CBD(0.1-10μg/mL)具有抗性,而T.denticolaATCC35405对所有测试的植物大麻素具有抗性(CBD,大麻酚[CBN],四氢大麻酚[THC])。RNAseq发现总共392个DenticolaATCC35405基因具有CBD响应性。通过qRT-PCR独立验证这些基因的所选子集。发现通过两种方法差异激活的基因包括几个参与转录调节和毒素控制的基因。抑制的基因包括几种参与趋化性和蛋白水解的基因。
结论:口腔螺旋体,与其他牙周细菌不同,对生理剂量的植物大麻素具有抗性。对CBD诱导的转录组变化的研究提供了对这种重要牙周病原体抗性机制的了解。应在报告的大麻使用者对牙周炎的易感性增加的背景下考虑这些发现。
The use of cannabis, which contains multiple antimicrobials, may be a risk factor for periodontitis. We hypothesized that multiple oral spirochetes would be phytocannabinoid-resistant and that cannabidiol (CBD) would act as an environmental stressor to which Treponema denticola would respond transcriptionally, thereby providing first insights into spirochetal survival strategies.
Oral spirochete growth was monitored spectrophotometrically in the presence and absence of physiologically relevant phytocannabinoid doses, the transcriptional response to phytocannabinoid exposure determined by RNAseq, specific gene activity fluxes verified using qRT-PCR and orthologues among fully sequenced oral spirochetes identified.
Multiple strains of oral treponemes were resistant to CBD (0.1-10 μg/mL), while T. denticola ATCC 35405 was resistant to all phytocannabinoids tested (CBD, cannabinol [CBN], tetrahydrocannabinol [THC]). A total of 392 T. denticola ATCC 35405 genes were found to be CBD-responsive by RNAseq. A selected subset of these genes was independently verified by qRT-PCR. Genes found to be differentially activated by both methods included several involved in transcriptional regulation and toxin control. Suppressed genes included several involved in chemotaxis and proteolysis.
Oral spirochetes, unlike some other periodontal bacteria, are resistant to physiological doses of phytocannabinoids. Investigation of CBD-induced transcriptomic changes provided insight into the resistance mechanisms of this important periodontal pathogen. These findings should be considered in the context of the reported enhanced susceptibility to periodontitis in cannabis users.