| Background:Burns is a common and frequently-occurring disease in clinical, of which the severity depends on the scope and depth of the damaged tissue. Shengfu Oil(SFO) is saprophytic muscle, anti-inflammatory, analgesic and can be used for treating ? degree burn wounds, which is composed of sixteen traditional drugs including radix scutellariae, olibanum, etc. Olibanum, the main compontent of SFO which is from the gum resin of Boswellia carterii, has demonstrated its anti-inflammatory and antioxidative activities, and these results provide a very interesting potential source of Traditional Chinese medicine for wound healing.Aim:This thesis investigated the scarless repairment effect and main pharmacodynamic studies of SFO on wounds of full-thickness scald in rabbits. In addition, the compositions of volatile oil from olibanum and the extractions of volatile oil from olibanum with different solventsis had been described. Anti-inflammatory and analgesic effects are compared among ?-Pinene, Linalool and 1-Octanol. At the same time, these results would provide basic experimental data to the studies on the mechanisms of the scarless repairment effect on wounds of full-thickness scald and the clinical application of SFO.Methods:1, Established the model of full-thickness scald wounds in rabbits by water vapor. Wounds with area of 6 cm2 were reproduced on both sides of the back in 36 rabbits. These rabbits were divided into sesame oil(S1), Shengfu oil(S2), and mupirocin ointment(M) groups, with 12 rabbits in each group. Both wounds of each rabbit in the three groups were respectively treated with S1, S2, and M, in dose of 0.15 mL/cm2, 2-3 times per day. The general condition of wounds was observed on post scald day(PSD) 1, 7, 14, and 43, the three rabbits of each group were sacrificed on PSD 1, 7, 14, and 43. The wounded tissues were subjected to histomorphological study with H&E staining. The protein expressions of ?-catenin, delta-like homologue 1(Dlk1), and cyclooxygenase-2(COX-2) were observed with immunohistochemistry staining for the other part of wounded tissues and were quantitated with the use of multispectral analysis software. Data were processed with two-way analysis of variance.2, The infiammatory model was made by dimethylbenzene-induced edema of mice ear, and the analgesic model was made by an injection of formalin into the plantar region of the right hind paws, followed by smearing different drugs or equal volume of 0.9% NaCl. Then, both ear edema and paw edema were measured at 0, 1, 2, 4, 8, 12 h and 0, 1, 4, 12 h, respectively. Data were processed with two-way analysis of variance.3, GC-MS was used to analyze the components of the essential oil from olibanum using different solvents. The contents of ?-Pinene, Linalool and 1-Octanol were determined by using internal standard method. The inflammatory model was made by dimethylbenzene-induced edema of the right ear, and the analgesic model was made by an injection of formalin into the plantar region of the right hind paws, followed by smearing different drugs or equal volume of 0.9% NaCl. Then, both ear edema and paw edema were measured at 0, 0.15, 1, 2, 4, 8, 12 h and 0, 4, 8, 12 h, respectively. Data were processed with two-way analysis of variance. Mice of each group were sacrificed. The foot skin tissues were subjected to histomorphological study with H&E staining. The protein expressions of COX-2 were observed with immunohistochemistry staining for the other part of foot skin tissues and were quantitated with the use of multispectral analysis software. Wound healing assay was established to study the effects of Olibanum and SFO on wound healing.Results:1, SFO can promote wound healing quickly: ? The wound healing quality of rabbits in SFO group was better than that in the other two groups. ? Histomorphological observation showed that the structure of wounded tissues in SFO group was intact, including regenerated cutaneous appendages.? Immunohistochemical staining showed that the protein expressions of ?-catenin?Dlk1 in SFO group were higher than those in control in early wound repair, accompanying with lower expression of COX-2; the protein expressions of ?-catenin fell in the middle of wound repair; the protein expressions of ?-catenin and Dlk1 raised in the end of wound repair. ? Wound healing assay showed that SFO can promote NIH/3T3 cell migration.2, SFO exerts anti-inflammatory and analgesic effects: 4 hours after applied SFO in the ears of mice, the ear edema was reduced significantly compared with the mice in model. SFO can efficiently inhibit the inflammatory response induced by dimethylbenzene. 12 hours after applied SFO in the feet of mice, the paw edema was raised to 111% of control. SFO can potently inhibit hyperalgesia induced by an injection of formalin, and the effect is long-acting.3, Olibanum exerts anti-inflammatory and analgesic effects: ? The content of ?-Pinene, Linalool and 1-Octanol is 0.072 mg/mL, 0.038 mg/mL, 0.0593 mg/mL in olibanum oil extract(OOE) respectively, while is 0.0015 mg/mL, 3.69 mg/mL, 0.73 mg/mL in olibanum alcohol extract(OAE). ? Compared to model, OOE and OAE can reduce the ear edema obviously, while OWE has no effect on it. OOE, OAE and OWE potently inhibit hyperalgesia. ?-Pinene, Linalool, 1-Octanol and a mixture of all three can inhibit inflammation and hyperalgesia in mice. ? Histomorphological observation showed that the inflammatory cells infiltration is lighter in ?-Pinene, Linalool, 1-Octanol and mixture compared with model. Linalool and mixture are better than the other two groups. Immunohistochemical staining showed that the protein expressions of COX-2 reduced in ?-Pinene, Linalool, 1-Octanol and mixture, 87%, 86%, 73% and 91%, respectively. ?-Pinene, Linalool and mixture are better than 1-Octanol. ? Wound healing assay showed that OOE promoted NIH/3T3 cell migration.Conclusion:1, SFO can promote scarless wound healing by regulating the expressions of ?-catenin?Dlk1 and COX-2 and exerting anti-inflammatory and analgesic effects. ?-Pinene, Linalool and 1-Octanol are the main volatile oil from olibanum, which exert anti-inflammatory and analgesic effects by reducing the expression of COX-2 in injured tissues. Olibanum can promote wound healing by accelerating cell migration. |
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