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目的利用转绿色荧光蛋白(green fluorescent protein,GFP)嵌合小鼠的动物模型,研究粒细胞集落刺激因子(granulocyte colony-stimulating factor,G-CSF)骨髓动员在创面修复中的作用和机制。方法 C57骨髓嵌合小鼠随机分成骨髓动员组和对照组。骨髓动员组小鼠连续5 d予以皮下注射G-CSF,对照组小鼠给予相同剂量的生理盐水。在小鼠背部制作皮肤全层缺损伤,1.3 cm×1.3 cm大小。骨髓动员组继续给予G-CSF 3 d,对照组给予生理盐水。伤后每天观察各组动物创面大小及愈合情况。取创伤后第1天的创面组织,消化成单细胞悬液,流式检测GFP+细胞比例。取伤后第3天对照组和骨髓动员组创面组织制取石蜡切片,免疫组化检测β-catenin表达。结果骨髓动员组小鼠创面愈合增快为(12.5±0.9)d,对照组愈合时间为(18.3±0.8)d。骨髓动员组在伤后1 d创面局部GFP+细胞明显多于对照组,差异有统计学意义(P<0.05)。骨髓动员组创面处β-catenin在创缘组织中的表达明显增强,与对照组相比差异具有统计学意义(P<0.05),并部分呈现核表达。结论 G-CSF动员后,GFP+细胞大量聚集于创面,并促进创面β-catenin的表达和核转位,加快创面愈合。
OBJECTIVE: To investigate the role and mechanism of bone marrow mobilization by granulocyte colony-stimulating factor (G-CSF) in the repair of wounds using an animal model of green fluorescent protein (GFP) chimeric mice. Methods C57 bone marrow chimeric mice were randomly divided into mobilization group and control group. The bone marrow mobilization group mice were injected subcutaneously with G-CSF for 5 days, and the control group mice were given the same dose of saline. Full-thickness skin lesions were made on the back of the mouse, 1.3 cm × 1.3 cm in size. The bone marrow mobilization group was given G-CSF for 3 days, and the control group was given normal saline. After wounding, the wound size and healing of each group were observed daily. The wound tissue on day 1 after wounding was digested into single cell suspension and the proportion of GFP + cells was detected by flow cytometry. On the 3rd day after injury, paraffin sections were obtained from the wound surface of control group and bone mobilization group, and the expression of β-catenin was detected by immunohistochemistry. Results The healing rate of bone marrow mobilization group was (12.5 ± 0.9) d and that of control group was (18.3 ± 0.8) d. The number of local GFP + cells in bone mobilization group was significantly higher than that in control group on the 1st day after wounding, the difference was statistically significant (P <0.05). The expression of β-catenin in the wound surface of mobilized bone marrow group was significantly higher than that of the control group (P <0.05), and some of them showed nuclear expression. Conclusion After mobilization of G-CSF, GFP + cells accumulate in large numbers on the wound surface, promote the expression and nuclear translocation of β-catenin, and accelerate wound healing.