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International Journal of Legal Medicine

, Volume 126, Issue 5, pp 807–814 | Cite as

Cannabinoid receptor type 2 is time-dependently expressed during skin wound healing in mice

  • Ji-Long Zheng
  • Tian-Shui Yu
  • Xiao-Na Li
  • Yan-Yan Fan
  • Wen-Xiang Ma
  • Yu Du
  • Rui Zhao
  • Da-Wei Guan
Original Article

Abstract

Dynamic localization of CB2R and quantitative analysis of CB2R mRNA during skin wound healing in mice were performed. Co-localization of CB2R with F4/80 or α-SMA was detected by double-color immunofluorescence microscopy. A total of 110 male mice were divided into control, injury, and postmortem groups. Sixty-five mice were sacrificed, followed by sampling at 0.5 h–21 days post-injury. Five mice without incision were used as control. The other 40 mice that received incised wound were sacrificed at 5 days after injury. The samples were collected at 0 h–3 days postmortem. In the uninjured controls, CB2R immunoreactivity was detected in the epidermis, hair follicles, sebaceous glands, dermomuscular layer, and vascular smooth muscle. In the incision groups, polymorphonulcear cells, macrophages, and myofibroblasts showed positive staining for CB2R. Morphometrically, the average ratios of CB2R-positive cells were more than 50 % at 5 days post-wounding, whereas it was <50 % at the other posttraumatic intervals. The average ratios of CB2R-positive macrophages maximized at 3 days post-wounding, and the average ratios of CB2R-positive myofibroblasts peaked at 5 days post-wounding. The relative quantity of CB2R mRNA expression maximized at posttraumatic 5 days in comparison with control as detected by real-time PCR, with an average ratio of >4.10, which was also confirmed by Western blotting. There was no significant change for CB2R protein within 6 h postmortem and for mRNA within 3 h postmortem as compared with the control group. In conclusion, dynamic distribution and expression of CB2R suggest that CB2R is involved in modulating macrophages and myofibroblasts in response to inflammatory event and repair process in mouse skin wound healing, and CB2R is available as a marker for wound age determination.

Keywords

Wound age determination Skin incision CB2R Macrophage Myofibroblast Real-time PCR 

Notes

Acknowledgments

This study was financially supported in part by a grant from research funds for the Doctoral Program funded by the Ministry of Education of China (200801590020) and a grant funded by the National Natural Science Foundation of China (30271347).

Supplementary material

414_2012_741_Fig6_ESM.jpg (8 kb)
Supplemental Fig. 1

Average ratio of CB2R-positive cells in each group. a p < 0.05 (vs control group); b p < 0.05 (vs preceding group) (JPEG 7 kb)

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High-resolution image (TIFF 88 kb)
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Supplemental Fig. 2

The average ratios of CB2R to GAPDH at different postmortem intervals. a p < 0.05 (vs 0 h group and preceding group) (JPEG 9 kb)

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High-resolution image (TIFF 162 kb)
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Supplemental Fig. 3

Relative intensity of CB2R to GAPDH. The data of 0 h represent the results obtained from the control group. All values are expressed as the means ± SD (n = 5). *p < 0.05 (vs control group); **p < 0.05 (vs control group and preceding group) (JPEG 8 kb)

414_2012_741_MOESM3_ESM.tif (86 kb)
High-resolution image (TIFF 85 kb)
414_2012_741_Fig9_ESM.jpg (8 kb)
Supplemental Fig. 4

Relative intensity of CB2R to GAPDH. All values are expressed as the means ± SD (n = 5). *p < 0.05 (vs control group and preceding group) (JPEG 7 kb)

414_2012_741_MOESM4_ESM.tif (103 kb)
High-resolution image (TIFF 103 kb)
414_2012_741_Fig10_ESM.jpg (8 kb)
Supplemental Fig. 5

Relative quantity of CB2R mRNA by real-time PCR at different posttraumatic intervals. The data of 0 h represent the results obtained from the control group. All values represent the means ± SD (n = 5). *p < 0.05 (vs control group); **p < 0.05 (vs control group and preceding group) (JPEG 8 kb)

414_2012_741_MOESM5_ESM.tif (146 kb)
High-resolution image (TIFF 145 kb)
414_2012_741_Fig11_ESM.jpg (7 kb)
Supplemental Fig. 6

Relative quantity of CB2R mRNA by real-time PCR at different postmortem intervals. All values represent the means ± SD (n = 5). *p < 0.05 (vs control group and preceding group) (JPEG 7 kb)

414_2012_741_MOESM6_ESM.tif (88 kb)
High-resolution image (TIFF 87 kb)

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Ji-Long Zheng
    • 1
    • 2
  • Tian-Shui Yu
    • 1
    • 3
  • Xiao-Na Li
    • 4
  • Yan-Yan Fan
    • 1
  • Wen-Xiang Ma
    • 1
  • Yu Du
    • 2
  • Rui Zhao
    • 1
  • Da-Wei Guan
    • 1
  1. 1.Department of Forensic PathologyChina Medical University School of Forensic MedicineShenyangPeople’s Republic of China
  2. 2.Department of Forensic MedicineChina Criminal Police UniversityShenyangPeople’s Republic of China
  3. 3.Key Laboratory of Evidence ScienceChina University of Political Science and Law, Ministry of EducationBeijingPeople’s Republic of China
  4. 4.Department of ChemistryChina Medical University School of Basic MedicineShenyangPeople’s Republic of China

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