摘要
目的:观察局部应用胰岛素样生长因子Ⅰ对大鼠皮肤创面愈合的作用,为临床应用胰岛素样生长因子Ⅰ加速创面愈合建立理论和实验依据。方法:实验于2002-05/06在北京世纪坛医院动物实验室完成。①清洁级SD大鼠32只,雌雄不拘,体质量180～200g。随机分为胰岛素样生长因子Ⅰ组、胰岛素样生长因子Ⅰ+重组人生长激素组、重组人生长激素组、生理盐水组,每组8只。②在大鼠背部脊柱切除直径1.6～1.8cm的圆形皮肤全层缺损,创面压迫止血后各组分别给药。③胰岛素样生长因子Ⅰ组:每次每个创面给液量0.1mL,胰岛素样生长因子Ⅰ浓度1mg/L;胰岛素样生长因子Ⅰ+重组人生长激素组:每次每个创面给液量0.1mL,胰岛素样生长因子Ⅰ浓度1mg/L,重组人生长激素浓度0.2U/mL;重组人生长激素组:每次每个创面给液量0.1mL,重组人生长激素浓度0.2U/mL;生理盐水组:每次每个创面给生理盐水,给液量0.1mL。④分别于伤后1、4、7、11、14、17d创面取材,作组织切片观察。在取材同时剩余创面给药,药量同前,在伤后11～13d创面愈合后,14、17d取材为愈合部位组织。取材后即时以10%多聚甲醛固定10h,逐级脱水,作组织切片。⑤利用免疫组织化学和图像分析,研究局部应用胰岛素样生长因子Ⅰ、重组人生长激素后,创伤模型大鼠皮肤创面愈合过程中与创面愈合相关因素增殖细胞核抗原、Ⅰ型胶原之间的关系。结果:实验大鼠32只全部进入结果分析。①在局部应用外源性胰岛素样生长因子Ⅰ、重组人生长激素后,创面愈合过程中组织内Ⅰ型胶原的变化:伤后1d,胰岛素样生长因子Ⅰ组和胰岛素样生长因子Ⅰ+重组人生长激素组在胞浆里出现了Ⅰ型胶原表达,到伤后11d创面接近愈合及14d和17d创面愈合后在细胞间质中仍有增加的趋势。②增殖细胞核抗原的变化:在核内,胰岛素样生长因子Ⅰ组和胰岛素样生长因子Ⅰ+重组人生长激素组的增殖细胞核抗原表达量于4d达到高峰值,重组人生长激素组和生理盐水组伤后增殖细胞核抗原的表达量随时间的推移逐渐增加,在7d时达到最高后就开始降低,14、17d创面愈合后,其表达量也就下降到正常范围。③胰岛素样生长因子Ⅰ的变化:胰岛素样生长因子Ⅰ组和胰岛素样生长因子Ⅰ+重组人生长激素组伤后在胞浆和细胞间质胰岛素样生长因子Ⅰ的表达量就一直在高值,在重组人生长激素组和生理盐水组,伤后1d出现胰岛素样生长因子Ⅰ的表达,在7d时达到峰值,在伤后11d,表达量下降,14、17d创面愈合后,其表达量也就下降到正常范围。④组间采用t检验。结论:在创面应用胰岛素样生长因子Ⅰ能够促进成纤维细胞分裂增殖,合成分泌增加,是成纤维细胞增生的直接促进因子,能促进创面的愈合。创面应用的重组人生长激素促增殖作用不强。
AIM： To investigate the effect of type Ⅰ insulin like growth factor （IGF-Ⅰ） on the wound healing of rats, so as to provide theoretical and experimental evidence for clinical application of IGF-1 to accelerate the process of wound healing. METHODS： The experiment was conducted in the Experimental Animal Laboratory of Beijing Shijitan Hospital from May to June 2002. ①Thirty- two clean grade Sprague-Dawley rats in either gender with body mass of 180 to 200 g were selected and randomly divided into IGF-1 group, IGF-1 and recombinant human growth hormone （rhGH） group, rhGh group and normal saline （negative control） groupwith 8 rats in each group. ②The round wound models with diameter of 1.6 to 1.8 cm were made in the back skin of rats and given interventions after hemostasis by compression. ③ IGF-1 group： Every wound was given liquid of 0.1 mL with IGF-1 concentration of 1 mg/L; IGF-1 and rhGH group： The dosage was 0.1 mL with IGF-1 concentration of 1 mg/L and rhGH concentration of 0.2 U/mL; rhGH group： The dosage was 0.1 mL with rhGH concentration of 0.2 U/mL; Negative control group： 0.1 mL physiological saline was applied in every wound. ④The specimens in the wound site were harvested after 1, 4, 7, 11, 14 and 17 days. The same dosage medicament was applied in the residuary wounds. The healing tissue was sampled at 14 and 17 days after the wound site was healed after 11 and 13 days. The samples were fixed with 10% formalin for 10 hours, differentiated and dehydrated in alcohol.⑤ Immunohistochemistry （IHC） and Image analysis was employed to detect the relationship between the related factors of wound healing： Proliferating cell nuclear antigen （PCNA） and Type Ⅰ collagen in the process of wound healing after the application of ectogenous IGF-1 and rbGH. RESULTS： Thirty-two rats were involved in the result analysis. ①Changes of expression of type Ⅰ collagen after IGF-1 and rhGH were applied in wound： The expression of type Ⅰ collagen was found in the cytolymph in IGF-1 group, IGF-1 and rhGH group after 1 day of wound, the expression kept the trend of increase in 11, 14 and 17 days after wound. ②Changes of expression of PCNA： The expression of PCNA in the cell nucleus in the IGF-1 group, IGF-1 and rhGH group reached the peak value at day 4. In the rhGH group and negative control group, the expression of PCNA gradually increased and arrived the peak value at day 7, then reduced step by step to the normal range after wound healed at days 14 and 17. ③ Expression of IGF-1： In the IGF-1 group, IGF-1 and rhGH group, the expression of IGF-1 in the cytolymph and cell nucleus always kept the peak value. In the rhGH group and negative control group, the expression of IGF-1 was found after 1 day and arrived the peak value at day 7, reduced from day 11 and to the normal range after wound healed in 14 days and 17 days. ④The t-test was used in the comparison among different groups.CONCLUSION： Application of ectogenous IGF-1 in wound will help to the splitting, proliferation and excretion of fibroblast; it is the direct promoter for proliferation of fibroblast and will accelerate the process of wound healing. On the other hand, the effect of rhGH used in wound is not powerful.
出处
《中国临床康复》
CSCD
北大核心
2006年第45期71-74,共4页
Chinese Journal of Clinical Rehabilitation
