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目的在体观察重组人血小板源性生长因子(recombinant human platelet-derived growth factor,rhPDGF)促进糖尿病大鼠全层皮肤缺损创面修复可能涉及的细胞和分子机制,研究其可能涉及的信号通路。方法 26只糖尿病大鼠,每只动物背部制备4个全层皮肤缺损创面,选取其中52个创面,随机分成3组,即对照组,创面自然愈合;rhPDGF治疗组,创面rhPDGF用量为7.0μg/cm2;赋形剂组,创面用等量赋形剂凝胶。观察治疗后3、7和14 d创面肉芽形成、胶原沉积、再上皮化速率以及炎性细胞浸润情况,并采用免疫荧光和免疫组织化学技术观察创面周围和创面修复细胞内细胞外信号调节激酶1/2(extracellular signal-regulated kinase 1/2, ERK1/2)磷酸化和增殖细胞核抗原(proliferative cell nuclear antigen, PCNA)的表达。结果组织学观察,rhPDGF治疗组创面可见大量炎性细胞浸润,毛细血管胚芽及成纤维细胞明显多于另两组(P<0.05);胶原沉积明显,肉芽组织生长活跃,创面收缩显著,与对照组比较差异有统计学意义(P<0.05)。免疫学研究显示,应用rhPDGF 7~14 d后,rhPDGF治疗组ERK1/2明显强于对照组和赋形剂组(P<0.05);且损伤后3~7 d rhPDGF治疗组修复细胞PCNA的表达明显高于对照组和赋形剂组(P<0.05)。结论 rhPDGF促糖尿病大鼠创面愈合的作用部分是通过ERK1/2信号通路的磷酸化来完成的。
OBJECTIVE: To observe the cellular and molecular mechanisms involved in the repair of full-thickness skin defects in diabetic rats with recombinant human platelet-derived growth factor (rhPDGF) and to investigate the possible signaling pathways. Methods Twenty-six diabetic rats were randomly divided into three groups (control group). The wounds were healed naturally. In rhPDGF-treated group, the dosage of rhPDGF was 7.0 μg / cm2; excipient group, wounds with the same amount of excipient gel. Granulation formation, collagen deposition, re-epithelialization rate and inflammatory cell infiltration were observed at 3, 7 and 14 days after treatment. Immunofluorescence and immunohistochemistry were used to observe the changes of extracellular signal-regulated kinase 1 / 2 (extracellular signal-regulated kinase 1/2, ERK1 / 2) phosphorylation and proliferating cell nuclear antigen (PCNA) expression. Results Histological observation showed that a large number of inflammatory cells were infiltrated into the wound surface of rhPDGF group, and the number of capillary villus and fibroblasts was significantly higher than that of the other two groups (P <0.05). Collagen deposition was obvious and granulation tissue grew actively. The difference was statistically significant (P <0.05). Immunological studies showed that after 7 to 14 days of rhPDGF treatment, the ERK1 / 2 level in rhPDGF-treated group was significantly higher than that in control group and vehicle group (P <0.05), and the expression of PCNA in rhPDGF-treated group from 3 to 7 days after injury Significantly higher than the control group and vehicle group (P <0.05). Conclusion The effect of rhPDGF on wound healing in diabetic rats is partly mediated by phosphorylation of ERK1 / 2 signaling pathway.