Department of Plastic, Reconstructive and Aesthetic Surgery and Burn Unit, Faculty of Medicine, University of Porto and Centro Hospitalar São João, Porto, Portugal.
Department of Biomedicine-Pharmacology and Therapeutics Unit, Faculty of Medicine, University of Oporto, Porto, Portugal.
Fundam Clin Pharmacol. 2022 Feb;36(1):89-99. doi: 10.1111/fcp.12716. Epub 2021 Aug 19.
Previous studies showed that cannabinoid 2 (CB2) receptor is involved in skin inflammation, fibrogenesis and re-epithelialization in mice, indicating that this receptor may be implicated in wound healing. Thus, topical use of cannabinoids may have a role in local fibrotic and wound healing diseases such as scars or keloids. We investigate the effect of the CB2 selective receptor agonist (6aR,10aR)-3-(1,1-Dimethylbutyl)-6a,7,10,10a-tetrahydro-6,6,9-trimethyl-6H-dibenzo[b,d]pyran (JWH133) and the CB2 selective receptor antagonist (6-Iodo-2-methyl-1-[2-(4-morpholinyl)ethyl]-1H-indol-3-yl)(4-methoxyphenyl)-methanone (AM630), on primary cultures of human fibroblasts. Primary cultures of adult human fibroblasts were obtained from abdominal human skin samples. Fibroblasts pretreated with JWH133 and/or AM630 were stimulated with TGF-β (10 ng/ml). Fibroblast activation into myofibroblasts was quantified by the expression of alpha-smooth muscle actin (α-SMA) using Immunocytochemistry and Western Blot assays. Collagen content was quantified with the Sirius red staining assay. Upon human fibroblasts stimulation with TGF-β, a significant increase on α-SMA and CB2 receptor expression was observed. In these cells, JWH133 decreased α-SMA expression and collagen content. However, this effect was not observed in resting human fibroblasts. AM630 decreased α-SMA expression and collagen content in both resting and activated fibroblasts. This effect was time- and concentration-dependent with an IC value of 11 μM. The CB2 receptor appears to be involved in fibroblast repair during skin wound healing in humans, as TGF-β increases CB2 receptor expression and JWH133 produces an anti-fibrotic effect in human fibroblasts. AM630 also showed an anti-fibrotic effect hypothesizing that other cannabinoid receptors, such as TRPV, may be involved in this response.
先前的研究表明，大麻素 2 型（CB2）受体参与了小鼠皮肤炎症、纤维化和再上皮化，这表明该受体可能与伤口愈合有关。因此，大麻素的局部应用可能在疤痕或瘢痕疙瘩等局部纤维化和伤口愈合疾病中发挥作用。我们研究了 CB2 选择性受体激动剂（6aR，10aR）-3-（1,1-二甲基丁基）-6a，7,10,10a-四氢-6,6,9-三甲基-6H-二苯并[b，d]吡喃（JWH133）和 CB2 选择性受体拮抗剂（6-碘-2-甲基-1-[2-（4-吗啉基）乙基]-1H-吲哚-3-基]（4-甲氧基苯基）-甲酮（AM630）对原代培养的人成纤维细胞的影响。从腹部人皮肤样本中获得原代培养的成人成纤维细胞。用 JWH133 和/或 AM630 预处理成纤维细胞，然后用 TGF-β（10ng/ml）刺激。用免疫细胞化学和 Western Blot 检测α-平滑肌肌动蛋白（α-SMA）的表达来量化成纤维细胞向肌成纤维细胞的激活。用天狼猩红染色法定量胶原蛋白含量。在 TGF-β刺激人成纤维细胞后，观察到α-SMA 和 CB2 受体表达显著增加。在这些细胞中，JWH133 降低了α-SMA 的表达和胶原蛋白含量。然而，在静止的人成纤维细胞中未观察到这种作用。AM630 降低了静止和激活的成纤维细胞中α-SMA 的表达和胶原蛋白含量。这种作用具有时间和浓度依赖性，IC 值为 11μM。CB2 受体似乎参与了人类皮肤伤口愈合过程中的成纤维细胞修复，因为 TGF-β增加了 CB2 受体的表达，JWH133 对人成纤维细胞产生抗纤维化作用。AM630 也表现出抗纤维化作用，假设其他大麻素受体，如 TRPV，可能参与了这种反应。
