International Journal of Legal Medicine

, Volume 125, Issue 4, pp 549–558 | Cite as

Time-dependent expression and distribution of monoacylglycerol lipase during the skin-incised wound healing in mice

  • Wen-Xiang Ma
  • Tian-Shui Yu
  • Yan-Yan Fan
  • Shu-Tao Zhang
  • Peng Ren
  • Shuai-Bo Wang
  • Rui Zhao
  • Jing-Bo Pi
  • Da-Wei Guan
Original Article


The study investigated the expression of monoacylglycerol lipase (MGL) during the skin-incised wound healing in mice and applicability of the time-dependent expression of MGL to wound age determination by immunofluorescent staining, Western blotting, and real-time PCR. Furthermore, cell types were identified by double immunofluorescence. A total of 45 BALB/c male mice were used in this study. After a 1.5-cm-long incision in the central dorsum skin, mice were killed at intervals ranging from 6 h to 14 days, followed by the sampling of wound margin. In the control, there was a low-level expression of MGL in the epidermis, hair follicles, and glandulae sebaceae. In the injured skin, MGL immunoreactivity was mainly detected in the neutrophils, macrophages, and myofibroblasts. Morphometrically, the average ratios of MGL-positive cells were more than 50% at 5 and 7 days post-wounding, whereas it was <50% at the other posttraumatic intervals. By Western blotting analysis, the average ratio of MGL protein expression was highest at 5 days after injury, which had a ratio of >2.30. Similarly, the relative quantity of MGL mRNA expression maximized at posttraumatic 5 days in comparison with control as detected by real-time PCR, with an average ratio of >2.54. In conclusion, MGL expression is detected in neutrophils, macrophages, and myofibroblasts and significantly up-regulated, suggesting that it may play roles in response to inflammation during skin-incised wound healing. From the viewpoint of forensic pathology, MGL detection is applicable to skin wound age determination.


Wound age determination Skin wound MGL Macrophage Myofibroblast Real-time PCR 



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


  1. 1.
    Marrs W, Stella N (2009) Measuring endocannabinoid hydrolysis: refining our tools and understanding. AAPS J 11:307–311PubMedCrossRefGoogle Scholar
  2. 2.
    Duncan M, Thomas AD, Cluny NL, Patel A, Patel KD, Lutz B, Piomelli D, Alexander SP, Sharkey KA (2008) Distribution and function of monoacylglycerol lipase in the gastrointestinal tract. Am J Physiol Gastrointest Liver Physiol 295:G1255–G1265PubMedCrossRefGoogle Scholar
  3. 3.
    Puffenbarger RA (2005) Molecular biology of the enzymes that degrade endocannabinoids. Curr Drug Targets CNS Neurol Disord 4:625–631PubMedCrossRefGoogle Scholar
  4. 4.
    Karlsson M, Reue K, Xia YR, Lusis AJ, Langin D, Tornqvist H, Holm C (2001) Exon–intron organization and chromosomal localization of the mouse monoglyceride lipase gene. Gene 272:11–18PubMedCrossRefGoogle Scholar
  5. 5.
    Blankman JL, Simon GM, Cravatt BF (2007) A comprehensive profile of brain enzymes that hydrolyze the endocannabinoid 2-arachidonoylglycerol. Chem Biol 14:1347–1356PubMedCrossRefGoogle Scholar
  6. 6.
    Singer AJ, Clark RA (1999) Cutaneous wound healing. N Engl J Med 341:738–746PubMedCrossRefGoogle Scholar
  7. 7.
    Guan DW, Ohshima T, Kondo T (2000) Immunohistochemical study on Fas and Fas ligand in skin wound healing. Histochem J 32:85–91PubMedCrossRefGoogle Scholar
  8. 8.
    Kondo T, Ohshima T (1996) The dynamics of inflammatory cytokines in the healing process of mouse skin wound: a preliminary study for possible wound age determination. Int J Leg Med 108:231–267CrossRefGoogle Scholar
  9. 9.
    Dreßler J, Bachmann L, Kasper M, Hauck JG, Müller E (1997) Time dependence of the expression ICAM (CD-54) in human skin wound. Int J Leg Med 110:299–304CrossRefGoogle Scholar
  10. 10.
    Hayashi T, Ishida Y, Kimura A, Takayasu T, Eisenmenger W, Kondo T (2004) Forensic application of VEGF expression to skin wound age determination. Int J Leg Med 118:320–325CrossRefGoogle Scholar
  11. 11.
    Kondo T, Ohshima T, Eisenmenger W (1999) Immunohistochemical and morphometrical study on the temporal expression of interleukin-1α(IL-1α) in human skin wounds for forensic wound age determination. Int J Leg Med 112:249–252CrossRefGoogle Scholar
  12. 12.
    Kondo T, Ohshima T, Mori R, Guan DW, Ohshima K, Eisenmenger W (2002) Immunohistochemical detection of chemokines in human skin wounds and its application to wound age determination. Int J Leg Med 116:87–91CrossRefGoogle Scholar
  13. 13.
    Dinh TP, Carpenter D, Leslie FM, Freund TF, Katona I, Sensi SL, Kathuria S, Piomelli D (2002) Brain monoglyceride lipase participating in endocannabinoid inactivation. Proc Natl Acad Sci USA 99:10819–10824PubMedCrossRefGoogle Scholar
  14. 14.
    Gulyas AI, Cravatt BF, Bracey MH, Dinh TP, Piomelli D, Boscia F, Freund TF (2004) Segregation of two endocannabinoid-hydrolyzing enzymes into pre- and postsynaptic compartments in the rat hippocampus, cerebellum and amygdala. Eur J Neurosci 20:441–458PubMedCrossRefGoogle Scholar
  15. 15.
    Comelli F, Giagnoni G, Bettoni I, Colleoni M, Costa B (2007) The inhibition of monoacylglycerol lipase by URB602 showed an anti-inflammatory and anti-nociceptive effect in a murine model of acute inflammation. Br J Pharmacol 152:787–794PubMedCrossRefGoogle Scholar
  16. 16.
    Jhaveri MD, Richardson D, Chapman V (2007) Endocannabinoid metabolism and uptake: novel targets for neuropathic and inflammatory pain. Br J Pharmacol 152:624–632PubMedCrossRefGoogle Scholar
  17. 17.
    Hohmann AG, Suplita RL, Bolton NM, Neely MH, Fegley D, Mangieri R, Krey JF, Walker JM, Holmes PV, Crystal JD, Duranti A, Tontini A, Mor M, Tarzia G, Piomelli D (2005) An endocannabinoid mechanism for stress-induced analgesia. Nature 435:1108–1112PubMedCrossRefGoogle Scholar
  18. 18.
    Desroches J, Guindon J, Lambert C, Beaulieu P (2008) Modulation of the anti-nociceptive effects of 2-arachidonoylglycerol by peripherally administered FAAH and MGL inhibitors in a neuropathic pain model. Br J Pharmacol 155:913–924PubMedCrossRefGoogle Scholar
  19. 19.
    McPartland JM (2008) Expression of the endocannabinoid system in fibroblasts and myofascial tissues. J Bodyw Mov Ther 12:169–182PubMedCrossRefGoogle Scholar
  20. 20.
    Kondo T (2007) Timing of skin wounds. Leg Med 9:109–114CrossRefGoogle Scholar
  21. 21.
    Mori R, Kondo T, Nishie T, Ohshima T, Asano M (2004) Impairment of skin wound healing in beta-1,4-galactosyltransferase-deficient mice with reduced leukocyte recruitment. Am J Pathol 164:1303–1314PubMedCrossRefGoogle Scholar
  22. 22.
    Oka S, Wakui J, Ikeda S, Yanagimoto S, Kishimoto S, Gokoh M, Nasui M, Sugiura T (2006) Involvement of the cannabinoid CB2 receptor and its endogenous ligand 2-arachidonoylglycerol in oxazolone-induced contact dermatitis in mice. J Immunol 177:8796–8805PubMedGoogle Scholar
  23. 23.
    Facchinetti F, Del Giudice E, Furegato S, Passarotto M, Leon A (2003) Cannabinoids ablate release of TNF alpha in rat microglial cells stimulated with lypopolysaccharide. Glia 41:161–168PubMedCrossRefGoogle Scholar
  24. 24.
    Jordà MA, Verbakel SE, Valk PJ, Vankan-Berkhoudt YV, Maccarrone M, Finazzi-Agrò A, Löwenberg B, Delwel R (2002) Hematopoietic cells expressing the peripheral cannabinoid receptor migrate in response to the endocannabinoid 2-arachidonoylglycerol. Blood 99:2786–2793PubMedCrossRefGoogle Scholar
  25. 25.
    Yu TS, Cheng ZH, Li LQ, Zhao R, Fan YY, Du Y, Ma WX, Guan DW (2010) The cannabinoid receptor type 2 is time-dependently expressed during skeletal muscle wound healing in rats. Int J Leg Med 124:397–404CrossRefGoogle Scholar
  26. 26.
    Di Marzo V, Bisogno T, De Petrocellis L, Melck D, Orlando P, Wagner JA, Kunos G (1999) Biosynthesis and inactivation of the endocannabinoid 2-arachidonoylglycerol in circulating and tumoral macrophages. Eur J Biochem 264:258–267PubMedCrossRefGoogle Scholar
  27. 27.
    Walter L, Dinh T, Stella N (2004) ATP induces a rapid and pronounced increase in 2-arachidonoylglycerol production by astrocytes, a response limited by monoacylglycerol lipase. J Neurosci 24:8068–8074PubMedCrossRefGoogle Scholar
  28. 28.
    Cecchi R (2010) Estimating wound age: looking into the future. Int J Leg Med 124:523–536CrossRefGoogle Scholar
  29. 29.
    Kagawa S, Matsuo A, Yagi Y, Ikematsu K, Tsuda R, Nakasono I (2009) The time-course analysis of gene expression during wound healing in mouse skin. Leg Med (Tokyo) 11:70–75Google Scholar
  30. 30.
    Sun JH, Wang YY, Zhang L, Gao CR, Zhang LZ, Guo Z (2010) Time-dependent expression of skeletal muscle troponin I mRNA in the contused skeletal muscle of rats: a possible marker for wound age estimation. Int J Leg Med 124:27–33CrossRefGoogle Scholar
  31. 31.
    Ohshima T, Sato Y (1998) Time-dependent expression of interleukin-10 (IL-10) mRNA during the early phase of skin wound healing as a possible indicator of wound vitality. Int J Leg Med 111:251–255CrossRefGoogle Scholar
  32. 32.
    Bai R, Wan L, Shi M (2008) The time-dependent expressions of IL-1beta, COX-2, MCP-1 mRNA in skin wounds of rabbits. Forensic Sci Int 175:193–197PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Wen-Xiang Ma
    • 1
  • Tian-Shui Yu
    • 1
  • Yan-Yan Fan
    • 1
  • Shu-Tao Zhang
    • 1
  • Peng Ren
    • 1
  • Shuai-Bo Wang
    • 1
  • Rui Zhao
    • 1
  • Jing-Bo Pi
    • 2
  • Da-Wei Guan
    • 1
  1. 1.Department of Forensic PathologyChina Medical University School of Forensic MedicineShenyangPeople’s Republic of China
  2. 2.Division of Translational BiologyThe Hamner Institutes for Health SciencesResearch Triangle ParkUSA

Personalised recommendations