Department of Woman, Child and General and Specialist Surgery, University of Campania "Luigi Vanvitelli".
Department of Experimental Medicine, University of Campania "Luigi Vanvitelli", Naples, Italy.
J Pediatr Gastroenterol Nutr. 2020 Nov;71(5):633-640. doi: 10.1097/MPG.0000000000002863.
In this study, we investigated the role of the cannabinoid receptor type 2 (CB2) in the bone loss associated with celiac disease (CD) evaluating the effect of its pharmacological modulation on osteoclast activity. We previously demonstrated a significant association between the CB2 Q63R variant and CD, suggesting it as a possible disease biomarker. Moreover, CB2 stimulation is beneficial for reducing osteoclast activity in several bone pathologic conditions.
In vitro osteoclasts (OCs) were differentiated from peripheral blood mononuclear cells of healthy donors, CD children at diagnosis and after 1 year of gluten-free diet (GFD) and characterized by real-time PCR and western blot for the expression of CB2 and specific osteoclastic markers, TRAP and Cathepsin K. TRAP assay and Bone Resorption assay were performed to evaluate osteoclast activity before and after 48 h exposure to CB2 selective drugs (JWH-133 and AM630) and Vitamin D.
We found in CD patients an osteoclast hyperactivation and low levels of CB2. CB2 stimulation with JWH-133 agonist is more effective than Vitamin D in reducing osteoclast activity whereas CB2 blockade with AM630 increases osteoclast activation. The anti-osteoporotic effect of JWH-133 decreases when used in co-treatment with vitamin D. GFD reduces osteoclast activity without restore CB2 expression.
CB2 could be a molecular marker to predict the risk of bone alterations in CD and a pharmacological target to reduce bone mass loss in patients who need a direct intervention on bone metabolism, in addition to the GFD.
在这项研究中,我们研究了大麻素受体 2 型(CB2)在乳糜泻(CD)相关骨丢失中的作用,评估了其药理学调节对破骨细胞活性的影响。我们之前证明了 CB2 Q63R 变体与 CD 之间存在显著关联,表明其可能是一种疾病生物标志物。此外,CB2 刺激对减少几种骨病理条件下的破骨细胞活性有益。
体外破骨细胞(OC)从健康供体的外周血单核细胞、诊断时的 CD 儿童和无麸质饮食(GFD)治疗 1 年后分化,并通过实时 PCR 和 Western blot 对 CB2 和特定破骨细胞标志物 TRAP 和组织蛋白酶 K 的表达进行鉴定。在 CB2 选择性药物(JWH-133 和 AM630)和维生素 D 暴露 48 小时前后进行 TRAP 测定和骨吸收测定,以评估破骨细胞活性。
我们发现 CD 患者存在破骨细胞过度激活和 CB2 水平降低。与维生素 D 相比,JWH-133 激动剂刺激 CB2 更能有效降低破骨细胞活性,而 AM630 阻断 CB2 则增加破骨细胞激活。JWH-133 的抗骨质疏松作用在与维生素 D 联合治疗时会降低。GFD 降低破骨细胞活性,但不能恢复 CB2 表达。
CB2 可能是预测 CD 骨改变风险的分子标志物,也是一种药理靶点,可以减少需要直接干预骨代谢的患者的骨量丢失,除了 GFD 之外。