小鼠皮肤切创愈合过程中Ⅱ型大麻素受体的表达

The time-dependent exprenssion of CB2R during skin incised wound healing in mice

目的观察皮肤切创愈合过程中CB2R表达变化的时间规律性。方法健康小鼠50只,随机分为10组,建立皮肤切创模型,应用免疫组织化学技术及Western bloting方法检测切创后不同时间段小鼠背部正中皮肤CB2R的表达。结果免疫组织化学结果显示,对照组CB2R表达于表皮、毛囊、皮脂腺、皮肌层、血管平滑肌、神经纤维外膜及神经纤维束束膜;伤后6h损伤区及周边区多核粒细胞少量表达CB2R,12～24h以单个核细胞阳性表达为主,3d以单个核细胞和梭形成纤维细胞表达为主,5～14d以梭形成纤维细胞为主。CB2R阳性细胞率于0h～3d逐渐升高,5d达最高峰,随后逐渐下降。经Western blotting检测显示,对照及各个损伤时间段都有CB2R阳性条带,其中5d为CB2R表达高峰。结论小鼠皮肤切创愈合过程中,CB2R于损伤区及周边区多核粒细胞、单个核细胞、梭形成纤维细胞中表达,并具有规律性,提示CB2R可能参与皮肤损伤愈合,并可用于皮肤损伤时间的推断。

Objective To investiage the time-dependent changes of CB2R in the expression of CB2R during the mice. Methods after the model of skin wound incised healing and skin wound was established in mice,a total of 50 mice were divided into ten groups randomly,the expression of CB2R in skin layer of the central dorsum was detected by immunohistochemical staining and Western blotting. Results CB2R immurore- activity was detected in epidemical layer, hair follicle, sebaceous gland, dermomuscular layer, vascular smooth muscle, nerve fiber adventitia and perimysium in the control skin; weak CB2R immunostaining was detected in polymorphonuclear cells （PMNs） in the wound and peripheral region in aged 6h, a large number of CB2R- positive mononuclear cells（MNCs） were identified from 12h to 24h, Afterwards ,the CB2R-positive ceils were mostly MNCs and spindle-shaped fibroblastic cells at 3d postinjury, and mainly in spindle-shaped fibroblastic, cells from 5d to 14d postinjury. The ratio of the CB2R-positive cells increased gradually in the wound specimens from Oh to 3d,and maximized at 5d,and decreased from 7d to 14d postinjury. The bands of CB2R expression were observed throughout the wound healing stages by Western blotting, and maximized at 5d. Conclution The results suggest that CB2R is expressed in PMNs, MNCs and spindle-shaped fibroblastic cells during healing process of skin wound in mice,and may be used as a newmarker for the wound age detemtination.