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The purpose of skin wound healing is a complex process, including hemostasis, inflammation, granulation tissue formation, epithelial covering, and the final tissue reconstruction. Ⅰ cannabinoid receptor (cannabinoid receptor type 1, CB1R) endogenous cannabinoid system (endocannabinoid system, ECs) one of the main members, it is a G protein-coupled receptors (G protein-coupled receptor GPCR), expressed mainly in the central nervous system, also part of the expression in the peripheral organs, such as liver, gastrointestinal. In this study, the identification of of CB1R different regions using two polyclonal antibodies detected in mouse skin damage CB1R expression and changes in the healing process, to explore the role of CB1R in the healing of skin damage and its application in injury time estimation possibilities. Materials and methods clean grade healthy adult Kunming mice 50, male, body weight 35 ~ 40g, were randomly divided into 10 groups. Incised skin is divided into nine segments group, another group of normal control group, each group of five mice. Ether gas inhalation anesthesia, the the back shearing treatment, conventional surgery disinfection to do a longitudinal President 1.5cm full-thickness skin incision in the back of the central, deep fascia. Each mouse was housed separately, sanitized and feed and tap water to prevent infection. After injury Oh, 6h, 12h, Id, 3d, 5d, 7d, 10d and 14d mice cervical dislocation were sacrificed to take the wound 1.5cm × 1.0cm skin tissue. Back shearing normal control group, the mice were anesthetized, not incised wounds were observed to 14d, were killed by cervical, take the same part of the same size of the skin, to prepare HE staining and immunohistochemical staining and Western blotting detection application. Using immunohistochemistry and Western blot analysis period 50 cases of skin incised wound CB1R changes in non-incised wound healing in mice skin tissue as control, x400 times under the microscope, the damage zone and the damage to the surrounding area of ??10 randomly selected a vision, to count the total number of cells, the number of positive cells and the rate of positive cells. Application Fluochem V2.0 software detects the average gray value of each time period, the measured data are expressed as mean ± standard deviation (x ± s) said. SPSS13.0 for Windows, one-way ANOVA, P lt; 0.05 was considered statistically significant. Results expressed in normal mouse skin, the epidermis, hair follicles, sebaceous glands and skin myometrium CB1R. Skin cut wounds 6h, polynuclear granulocytes did not express CBIR. After injury, 12-24h, part of the mononuclear cells CB1R positive. L-3d after injury CB1R positive cells formed mononuclear cells and shuttle fibroblast-like cell-based. After injury, 5-14d, CB1R expression of fibroblast-like cells formed mainly in the shuttle. Oh-ld, CB1R after injury rate of positive cells gradually increased. 3-5d after injury showed high expression state, 5d peaked. 7 days after injury-14d CB1R-positive cells decreased gradually. Groups with adjacent on the set of the total number of cells and the positive cell rate differences were statistically significant (P lt; 0.05). Western blot analysis showed that CB1R positive with normal skin. The incised skin after each period group CB1R expression intensity of certain laws. After injury, Oh-3d, positive bands gradually increased; 5d after injury, the CB1R peaked. After injury 7d-14d, positive bands remain relatively strong, but the downward trend. In addition to the average gray value in the the 0h normal control group showed no significant difference (P gt; 0.05), among other groups the average gray value differences were statistically significant (P lt; 0.05). Conclusions normal mouse skin epidermis, hair follicles, sebaceous glands and skin muscularis expression CB1R. Mice skin incised wound healing process in mononuclear cells and fibroblasts express CB1R prompted CB1R may be involved in the inflammatory response and the healing of skin damage. Mouse skin incised wound healing in the CB1R expression changed regularly in the healing process may be used for the incised skin damage time inference.
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