摘要
目的:探讨急性放射性皮肤溃疡形成早期酸性成纤维细胞生长因子及其受体的表达和意义。方法:实验于1999-06/2004-12在军事医学科学院完成。选取二级健康雌性Wistar大鼠80只,随机分为照射组40只、创伤组30只、正常对照组10只。照射组以60Coγ射线50Gy单次局部照射建立急性放射性皮肤溃疡动物模型,照射部位为双后大腿、臀部及全尾;创伤组于背部制作直径1.5cm单纯皮肤伤口动物模型;正常对照组未作任何处理。采用免疫组化、原位杂交等方法检测创面内酸性成纤维细胞生长因子及其受体的表达。400倍条件下光镜观察其相对含量,以视野内阳性细胞数量为准,分为阴性(视野内无阳性信号)、弱阳性(视野内可见10个以下阳性信号)、阳性(视野内可见11～30个阳性信号)、强阳性(视野内可见30个以上阳性信号)。结果:实验共纳入80只大鼠,进入结果分析73只,死亡7只。①照射组与创伤组大体观察:照射组照射后1～7d未出现异常反应,第9天出现尾根部红肿及足底轻度红肿,第28天尾根部与大腿内侧脱毛、糜烂及浅小溃疡达高峰,第35天伤口仍未愈合,溃疡周围可见新生上皮迟缓长入,第55天溃疡周围新生上皮部分覆盖创面,伤口有收缩趋势,但仍未愈合。创伤组皮肤伤口均经历炎症渗出、伤口收缩、瘢痕愈合阶段,伤后1～2d炎性渗出较多,第3天伤口开始收缩,伤后18d时伤口全部愈合。②照射组与创伤组基本病理学变化:照射组:溃疡前期表皮细胞及毛囊上皮肿胀、核固缩、碎裂,真皮及皮下组织出现充血性血管反应,胶原纤维肿胀、融解、断裂、水肿、排列紊乱,并有灶状表皮浅层缺失灶;溃疡期溃疡表面为坏死组织层,其下为增生不良的肉芽组织,其内极少见新生血管,胶原纤维变性、融解、断裂、水肿,可见成纤维细胞松散聚集形成的细胞团。创伤组:伤后1～3d,创面大量炎细胞、浆液和纤维素渗出,成纤维细胞数量增加,散在肉芽组织形成;伤后5～9d,肉芽组织增殖达到高峰,上皮细胞增殖旺盛;伤后11d,伤口逐渐愈合;伤后11～21d,肉芽组织减少,由瘢痕组织取代;伤后21～28d,成纤维细胞数量明显减少,胶原纤维逐渐增宽呈条束状。③酸性成纤维细胞生长因子及其受体的免疫组化及原位杂交结果:正常对照组:皮肤内无明显的酸性成纤维细胞生长因子及其受体的阳性信号。创伤组:伤后5～10d,创面肉芽组织内酸性成纤维细胞生长因子及其受体表达呈强阳性,创面愈合后表达迅速减弱。照射组:形成溃疡前(照射后1～11d)组织内酸性成纤维细胞生长因子及其受体表达逐渐增强,呈阳性;溃疡形成后(照射后14～28d)酸性成纤维细胞生长因子及其受体表达较创伤组伤后5～10d明显减弱。结论:急性放射性皮肤溃疡内酸性成纤维细胞生长因子及其受体表达的减弱可能与其愈合延迟有关,早期应用生长因子制剂可能有助于溃疡的愈合。
AIM： To investigate the expressions and significances of acidic fibroblast growth factor （a-FGF） and its receptor at the early stage of acute radiationinduced skin ulcers. METHODS： The experiment was finished in the Academy of Military Medical Sciences from June 1999 to December 2004. Eighty healthy second-degree female Wistar rats were enrolled in the study, .and randomized into three groups： radiation group （n=40）, trauma group （n=30） and normal control group （n=10）. In the radiation group, tat models of acute radiation-induced skin injury were made with 50 Gy ^60Coγ-rays on two lower limbs, buttock and tails. In the trauma group, models of skin wound at the back with the diameter of 1.5 cm were made. Rats in the normal control group did not receive any treatment. Immunohistochemieal staining and in situ hybridization were used to detect the expressions of a-FGF and its receptor in simple skin wounds and acute radiation-induced skin ulcers. The relative content was observed under light microscope, and divided into four degrees according to the number of positive cells： negative （none positive signals in sights）, weak positive （less than 10 positive signals in sights）, positive （10-30 positive signals in sights）, strong positive （more than 30 positive signals in sights）. RESULTS： Eighty rats were enrolled in the study, 7 died and 73 were involved in the analysis of results. ① Gross observation： In the radiation group, no abnormal reaction was shown from 1 to 7 days, red swelling appeared at 9 days on tails and sole of feet, hair abscission, erosion, small ulcers increased maximally at 28 days on tails and inside of lower limbs, neonate epithelia grow slowly at 35 days, the ulcers contracted and partially healed at 55 days; In the trauma group, the wounds healing stages included inflammation exudation,wound contraction, scar healing stages, inflammation exudation was rich from 1 to 2 days,wound contraction appeared at 3 days, and all wounds healed at 18 days after injury.② Basic pathological changes： In the radiation group,epithelia and hair follicle swelling, karyopyknosis, fragmentation, vascular congested reaction, collagen fibers swelling, rupture, disordered array were observed during the pre-uleer stage; During the ulcer stage,the superficial layer was necrotic tissues, below that granulation tissues proliferated weakly,few new vessels appeared,and scattered cellular balls consisted of fibroblasts were seen in lower layers.In the trauma group, large exudation of inflammatory cells and cellulose,and scattered granulation tissues showed from 1 to 3 days; the epithelia and granulation proliferated rapidly from 5 to 9 days; the wounds healed gradually from 11 to 21 days, granulation tissues decreased and substituted by sears, the number of fibroblasts decreased and collagen fibers widened gradually into zona fasciculata.③ Results of immunohistochemistry and in situ hybridization： There were no obvious positive signals of a-FGF and its receptor in normal skin;In the trauma group,the expressions of a-FGF and its receptor were strongly positive in granulation tissues from 5 to lO days after injury,and decreased rapidly after wound healing;while in the radiation group,the expressions of a-FGF and its receptor were increased gradually from 1 to 11 days after radiation, and became weaker in granulation tissues from 21 to 28 days after radiation as compared with those in the trauma group from 5 to 10 days after injury. CONCLUSION： The radiation-induced reduce of the expressions of a-FGF and its receptor may be relevant to the wound healing retardation of acute radiation-induced skin ulcers, early administration of growth factors may contribute to the healing of radiation-induced skin ulcers.
出处
《中国临床康复》
CSCD
北大核心
2005年第30期107-109,i0003,共4页
Chinese Journal of Clinical Rehabilitation
