Department of Pathology, Microbiology and Immunology, University of South Carolina School of Medicine, Columbia, SC, United States.
Front Immunol. 2019 Aug 21;10:1921. doi: 10.3389/fimmu.2019.01921. eCollection 2019.
Multiple sclerosis (MS) is a chronic and disabling disorder of the central nervous system (CNS) characterized by neuroinflammation leading to demyelination. Recently a combination of Δ9-tetrahydrocannabinol (THC) and Cannabidiol (CBD) extracted from Cannabis has been approved in many parts of the world to treat MS-related spasticity. THC+CBD combination was also shown to suppresses neuroinflammation, although the mechanisms remain to be further elucidated. In the current study, we demonstrate that THC+CBD combination therapy (10 mg/kg each) but not THC or CBD alone, attenuates murine experimental autoimmune encephalomyelitis (EAE) by reducing neuroinflammation and suppression of Th17 and Th1 cells. These effects were mediated through CB1 and CB2 receptors inasmuch as, THC+CBD failed to ameliorate EAE in mice deficient in CB1 and CB2. THC+CBD treatment also caused a decrease in the levels of brain infiltrating CD4+ T cells and pro-inflammatory molecules (IL-17, INF-γ, TNF-α, IL-1β, IL-6, and TBX21), while increasing anti-inflammatory phenotype such as FoxP3, STAT5b, IL-4, IL-10, and TGF-β. Also, the brain-derived cells showed increased apoptosis along with decreased percentage in G0/G1 phase with increased percentage in G2/M phase of cell cycle. miRNA microarray analysis of brain-derived CD4+ T cells revealed that THC+CBD treatment significantly down-regulated miR-21a-5p, miR-31-5p, miR-122-5p, miR-146a-5p, miR-150-5p, miR-155-5p, and miR-27b-5p while upregulating miR-706-5p and miR-7116. Pathway analysis showed that majority of the down-regulated miRs targeted molecules involved in cycle arrest and apoptosis such as CDKN2A, BCL2L11, and CCNG1, as well as anti-inflammatory molecules such as SOCS1 and FoxP3. Additionally, transfection studies involving miR-21 and use of mice suggested that while this miR plays a critical role in EAE, additional miRs may also be involved in THC+CBD-mediated attenuation of EAE. Collectively, this study suggests that combination of THC+CBD suppresses neuroinflammation and attenuates clinical EAE development and that this effect is associated with changes in miRNA profile in brain-infiltrating cells.
多发性硬化症（MS）是一种中枢神经系统（CNS）的慢性和致残性疾病，其特征为神经炎症导致脱髓鞘。最近，从大麻中提取的Δ9-四氢大麻酚（THC）和大麻二酚（CBD）的组合已在世界许多地方获得批准，用于治疗与 MS 相关的痉挛。THC+CBD 组合也被证明可以抑制神经炎症，尽管其机制仍有待进一步阐明。在目前的研究中，我们证明 THC+CBD 联合治疗（各 10mg/kg）而非单独的 THC 或 CBD 可通过减少神经炎症和抑制 Th17 和 Th1 细胞来减轻实验性自身免疫性脑脊髓炎（EAE）。这些作用是通过 CB1 和 CB2 受体介导的，因为 THC+CBD 未能改善 CB1 和 CB2 缺失的小鼠的 EAE。THC+CBD 治疗还导致大脑浸润的 CD4+T 细胞和促炎分子（IL-17、INF-γ、TNF-α、IL-1β、IL-6 和 TBX21）水平降低，同时增加抗炎表型，如 FoxP3、STAT5b、IL-4、IL-10 和 TGF-β。此外，大脑来源的细胞显示出凋亡增加，同时 G0/G1 期的百分比降低，G2/M 期的百分比增加。大脑来源的 CD4+T 细胞的 miRNA 微阵列分析显示，THC+CBD 治疗显著下调 miR-21a-5p、miR-31-5p、miR-122-5p、miR-146a-5p、miR-150-5p、miR-155-5p 和 miR-27b-5p，同时上调 miR-706-5p 和 miR-7116。通路分析表明，大多数下调的 miRs 靶向与细胞周期停滞和凋亡相关的分子，如 CDKN2A、BCL2L11 和 CCNG1，以及抗炎分子，如 SOCS1 和 FoxP3。此外，涉及 miR-21 的转染研究和使用 miR-21 缺失小鼠表明，虽然这种 miR 在 EAE 中发挥关键作用，但其他 miR 也可能参与 THC+CBD 介导的 EAE 衰减。总的来说，这项研究表明，THC+CBD 的组合抑制神经炎症并减轻临床 EAE 的发展，这种作用与大脑浸润细胞中 miRNA 谱的变化有关。
