摘要
目的了解感觉神经对皮肤损伤愈合中肉芽组织成纤维细胞表达的表皮生长因子(epidermal growth factor,EGF)、碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)蛋白表达的影响.方法在体实验中Wistar大鼠分为辣椒素组和对照组,对辣椒素组实验大鼠皮下注射辣椒素(capsaicin),以特异化学毁损感觉神经.停止注射辣椒素后一周行鼠背部皮肤全层切割伤,采用免疫组化结合图像分析,分别观察伤后3～12 d肉芽组织内EGF、bFGF和主要由感觉神经末梢释放的感觉神经肽P物质,对照组大鼠除不毁损感觉神经外,余处理和观察指标同辣椒素组.离体实验采用无菌肉芽组织培养的成纤维细胞,以蛋白印迹观察不同浓度(10 9～10-5 mol/L)P物质作用培养大鼠肉芽组织成纤维细胞后,上述生长因子的蛋白表达.结果肉芽组织P物质免疫阳性染色除分布于小血管周围、细胞间隙外,成纤维细胞胞浆也有较强的染色.辣椒素组化学去除感觉神经后肉芽组织P物质与EGF、bFGF蛋白质含量均较对照组减少,提示P物质可上调培养肉芽组织成纤维细胞EGF、bFGF的蛋白表达.结论皮肤创面愈合中成纤维细胞内源性EGF、bFGF蛋白表达与感觉神经释放的感觉神经肽P物质有关.
Objective To study the effect of sensory nerves on the expression of epidermal growth factor(EGF), basic fibroblast growth factor(bFGF) by fibroblasts in the granulation tissues in skin wound healing. Methods In vivo and in vitro studies were conducted. Fifty wistar rats were randomly divided into capsaicin group and control group. The rats in the capsaicin group were subcutaneously injected with neurotoxin dose of capsaicin(50 mg/kg for 3 days) to chemically destroy the sensory nerves. Full thickness skin incisive wound was made on the rat′s back on 8th day after last injection. The contents of substance P(SP), which was mainly released from afferent sensory nerve terminal, and EGF, bFGF in granulation tissue of skin lesion were determined using immunohistochemistry combined with computerized imaging analysis on 3rd, 6th, 9th, 12th days after the incision. In the control group no capsaicin was injected. In the in vitro study the fibroblasts from granulation tissue of rats were cultured with different doses of SP(10-9 ~ 10-5 mol/L), and the protein expression of EGF, bFGF by cultured fibroblasts was determined using Western blot. Results Immunoreactive SP was not only found around the small blood vessels and in the intercellular space, but also in the cytoplasm of fibroblasts in granulation tissue. SP content and the protein expression of EGF, bFGF in granulation tissue decreased in the capsaicin group, compared with those in the control group, which indicated that SP(10-7 mol/L) added into the culture medium for fibroblasts could upregulate the levels of EGF, bFGF expression. Conclusions Protein expression of EGF and bFGF by fibroblasts in granulation tissue in skin wound healing might be related with the SP released from sensory nerves.
出处
《中华皮肤科杂志》
CAS
CSCD
北大核心
2004年第10期578-580,共3页
Chinese Journal of Dermatology
基金
国家重点基础研究发展规划项目(G1999054024)
