皮肤创伤修复过程中TGF-β1及胶原表达变化的定量研究

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目的探讨皮肤创伤修复过程中TGF-β1的表达、成纤维细胞增殖、凋亡和胶原表达的动态变化及其意义。方法清洁成年雄性C57小鼠30只随机分为对照组和模型组,在小鼠的背部造成直径为0.4 cm的圆形全层皮肤缺损创面。于伤后1、4、7、14 d,取创面及周边的组织,常规制片HE染色,采用免疫组化染色观察创面及边缘TGF-β1和PCNA的表达,采用TUNEL法检测细胞凋亡及用天狼猩红染色方法显示胶原表达变化情况。结果 1TGF-β1的表达于伤后1 d组创缘处表皮细胞呈阳性,阳性率为19.87%±1.16%,成纤维细胞中表达较弱;伤后4 d组成纤维细胞中呈较强阳性,阳性率为37.15%±3.15%,伤后7 d组成纤维细胞阳性率达47.66%±3.71%;14 d组成纤维细胞呈弱阳性,阳性率为10.08%±0.12%。2PCNA于伤后1 d组阳性率为12.1%±1.12%;伤后4 d组阳性率为34.3%±2.31%,以成纤维细胞增殖为主;伤后7d组成纤维细胞增殖显著,阳性率达58.5%±4.73%;创后14 d组细胞增殖明显减弱,阳性率为6.9%±0.12%。3细胞凋亡检测示伤后1 d组皮肤创面及边缘细胞凋亡率为36.15±2.47%;伤后4 d凋亡率为27.15±2.30%,伤后7 d细胞凋亡率为11.71±0.98%,伤后14 d凋亡率达47.38%±2.16。4伤后1 d组胶原纤维的表达强度基本同对照组,伤后4 d胶原表达开始增高,7 d显著增高,于14 d胶原表达基本同对照组。结论在皮肤缺损创面修复过程中,TGF-β1的表达与成纤维细胞增殖及胶原合成呈正相关。 Objective To investigate the dynamic changes of TGF-β1, fibroblast proliferation, apoptosis and collagen expression in skin wound repair and its significance. Methods Thirty adult male C57 mice were randomly divided into control group and model group. A circular full - thickness skin defect with a diameter of 0.4 cm was created on the back of mice. At 1, 4, 7 and 14 days after wounding, the wounds and the surrounding tissues were taken for routine HE staining. The expressions of TGF-β1 and PCNA were detected by immunohistochemical staining. TUNEL method was used to detect the apoptosis and Sirius red staining showed changes in collagen expression. Results The expression of 1TGF-β1 was found on the wounds on the 1st day after wounding. The positive rate of 1TGF-β1 was 19.87% ± 1.16% and the expression of 1TGF-β1 was weak in fibroblasts. The positive rate was 37.15% ± 3.15%. The positive rate of fibroblasts on day 7 after injury was 47.66% ± 3.71%. On the 14th day, fibroblasts were weakly positive and the positive rate was 10.08% ± 0.12%. The positive rate of 2PCNA on day 1 after injury was 12.1% ± 1.12%; on day 4 after injury, the positive rate was 34.3% ± 2.31%, which was mainly fibroblast proliferation; on the 7th day after injury, the proliferation of fibroblasts was significant 58.5% ± 4.73%. After 14 days, the cell proliferation was significantly weakened, the positive rate was 6.9% ± 0.12%. The apoptosis rate of wounds and margins on the first day after wounding was 36.15 ± 2.47%. The apoptosis rate on the 4th day after injury was 27.15 ± 2.30%, and the apoptosis rate on the 7th day after injury was 11.71 ± 0.98 %, And the apoptotic rate reached 47.38% ± 2.16.4 on the 14th day after injury. The expression of collagen fibers in the first day after injury was almost the same as that in the control group. Collagen expression began to increase 4 days after injury and was significantly increased on the 7th day. Basic with the control group. Conclusion The expression of TGF-β1 is positively correlated with the proliferation of fibroblasts and collagen synthesis during the repair of wounds with skin defects.
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