Bort Alicia, Alvarado-Vazquez Perla A, Moracho-Vilrriales Carolina, Virga Kristopher G, Gumina Giuseppe, Romero-Sandoval Alfonso, Asbill Scott
1 Department of Biochemistry and Molecular Biology, School of Medicine, Alcalá de Henares, Madrid, Spain.
2 Department of Pharmaceutical and Administrative Sciences, Presbyterian College School of Pharmacy, Clinton, SC, USA.
Mol Pain. 2017 Jan;13:1744806916688220. doi: 10.1177/1744806916688220.
Background JWH015 is a cannabinoid (CB) receptor type 2 agonist that produces immunomodulatory effects. Since skin cells play a key role in inflammatory conditions and tissue repair, we investigated the ability of JWH015 to promote an anti-inflammatory and pro-wound healing phenotype in human primary skin cells. Methods Human primary keratinocytes and fibroblasts were stimulated with lipopolysaccharide. The mRNA expression of cannabinoid receptors was determined using RT-PCR. The effects of JWH015 (0.05, 0.1, 0.5, and 1 µM) in pro- and anti-inflammatory factors were tested in lipopolysaccharide-stimulated cells. A scratch assay, using a co-culture of keratinocytes and fibroblasts, was used to test the effects of JWH015 in wound healing. In addition, the topical and transdermal penetration of JWH015 was studied in Franz diffusion cells using porcine skin and LC-MS. Results The expression of CB1 and CB2 receptors (mRNA) and the production of pro- and anti-inflammatory factors enhanced in keratinocytes and fibroblasts following lipopolysaccharide stimulation. JWH015 reduced the concentration of major pro-inflammatory factors (IL-6 and MCP-1) and increased the concentration of a major anti-inflammatory factor (TGF-β) in lipopolysaccharide-stimulated cells. JWH015 induced a faster scratch gap closure. These JWH015'seffects were mainly modulated through both CB1 and CB2 receptors. Topically administered JWH015 was mostly retained in the skin and displayed a sustained and low level of transdermal permeation. Conclusions Our findings suggest that targeting keratinocytes and fibroblasts with cannabinoid drugs could represent a therapeutic strategy to resolve peripheral inflammation and promote tissue repair.
JWH015是一种能产生免疫调节作用的2型大麻素（CB）受体激动剂。由于皮肤细胞在炎症状态和组织修复中起关键作用，我们研究了JWH015在人原代皮肤细胞中促进抗炎和促伤口愈合表型的能力。方法：用脂多糖刺激人原代角质形成细胞和成纤维细胞。采用RT-PCR测定大麻素受体的mRNA表达。在脂多糖刺激的细胞中测试JWH015（0.05、0.1、0.5和1µM）对促炎和抗炎因子的影响。使用角质形成细胞和成纤维细胞的共培养物进行划痕试验，以测试JWH015在伤口愈合中的作用。此外，在Franz扩散池中使用猪皮和LC-MS研究JWH015的局部和透皮渗透情况。结果：脂多糖刺激后，角质形成细胞和成纤维细胞中CB1和CB2受体（mRNA）的表达以及促炎和抗炎因子的产生均增强。JWH015降低了脂多糖刺激细胞中主要促炎因子（IL-6和MCP-1）的浓度，并增加了主要抗炎因子（TGF-β）的浓度。JWH015诱导划痕间隙闭合更快。JWH015的这些作用主要通过CB1和CB2受体调节。局部给药的JWH015大多保留在皮肤中，并表现出持续且低水平的透皮渗透。结论：我们的研究结果表明，用大麻素药物靶向角质形成细胞和成纤维细胞可能是解决外周炎症和促进组织修复的一种治疗策略。
