摘要
目的:研究大鼠急性β射线皮肤损伤创面愈合过程中转化生长因子-1(transforming growth factorβ1,TGF-β1)及其受体(TβRⅠ)表达的动态变化,为深入探讨急性β射线皮肤损伤创面愈合的机制提供理论依据。
方法:雌性SD清洁大鼠54只,随机分为3组。照射组,n=24,直线加速器产生β射线(45Gy)单次照射大鼠臀部皮肤40mm×40mm,建立急性β射线皮肤损伤模型;烧伤组,n=24,将直径30mm铜柱于100℃沸水中放置15分钟,取出后在无外界压力下置于大鼠臀部皮肤8秒,建立深Ⅱ°热烧伤模型;对照组,n=6,正常大鼠,取不同时期创面皮肤组织,采用免疫组织化学方法(SP法)及原位逆转录聚合酶链反应(RT-PCR)和图像分析等方法,检测不同时相点各组大鼠创面组织中TGF-β1/TβRⅠ和TGF-β1mRNA/TβRⅠmRNA的转录及表达水平.
结果:①在照射后四周照射野内出现皮肤溃疡,之后逐渐扩大、融合、加深。②皮肤受照射区多种细胞,特别是溃疡床表皮细胞、成纤维细胞及血管内皮细胞中的转录及表达均较正常皮肤明显增强。③与单纯烧伤组比较,照射组的溃疡床的TGF-β1及TβRⅠ阳性细胞数量明显减少,阳性强度减少不明显,在溃疡出现后前三周内平均积分吸光度(MOD)值有统计学意义。④RT-PCR显示TGF-β1mRNA/TβRⅠmRNA表达的变化基本与免疫组化相似。
结论:辐射照射损伤的皮肤组织细胞合成分泌TGF-β1的功能在溃疡形成前被轻度激活,但在溃疡形成后虽然阳性程度仍较强,但阳性细胞数量明显少于单纯烧伤组,表明急性β射线皮肤损伤创面愈合过程中TGF-β1及TβRⅠ的表达水平降低,不能形成高峰值,可能与急性β射线皮肤损伤创面难愈合的分子机制之一。
Objective:The dynamic expression of TGF-β1/TβRⅠwere investigated Inβ-ray radiation injury of skin wounds,for in-depth study ofβ-ray skin injury wound healing mechanism difficult to provide a theoretical basis.
Methods:Fifty-four male SD were divided into 3 groups.The model of acute radiation of skin wound in rats replicated by 45 Gyβ-ray to skin radiation (R,n=24);The copper column with 30 mm diameter will be boiling water for 15 minutes after removal in the absence of outside pressure placed under burns(B,n=24);and 6 normal rats were served as normal6control group (N,n=6).From different pererent periods skin wounds,the expression of TGF-β1 and TβRⅠand TGF-β1mRNA/ TβRⅠmRNA were respectively semi-quantitative checked by means of immunohistochemical technique SP and reverse transcription PCR(RT-PCR).
Results:(1)Skin ulcers were found on week 4 after irradiation and they were enlarged and deepened gradually during the observation period .(2)In the irradiated skin ,especially in the epidermal cells ,fibroblasts and endothelial cells of the ulcer beds,the expression of TGF-β1 and TβRⅠwas higher than that in normal skin.(3)Compared with that in the surgical wound beds ,the number of TGF-β1 and TβRⅠpositive cells in radiation ulcer beds was obviously decreased .but TGF-β1 and TβRⅠpositive intensity was only slightly or not affected.(4)The results of RT-PCR for TGF-β1mRNA/ TβRⅠmRNA were in line with the results of the immunohistochemistry.
Concludion:The continuing low expression of TGF-β1 and TβRⅠmay be partially involved in the mechanism of retarded of acuteβ-ray radiation injury in skin wounds.
引文
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