摘要
目的探讨糖尿病(DM)合并深二度烫伤创面中成纤维细胞(FB)主要生物学功能及其超微结构的变化,以及该变化与 DM 创面愈合延迟的病理关系。方法将80只 SD 大鼠随机分为正常对照组和糖尿病组,各组分别于烫伤前及伤后3、7、14、21 d 采集创面皮肤组织标本。用流式细胞仪检测创面真皮组织中 FB 的增值周期;采用氯胺 T 氧化法测定创面组织羟脯氨酸(OHP)含量;应用 ELISA 技术检测Ⅰ/Ⅲ胶原比例;SP 免疫组化法检测α-肌动蛋白(α-SMA)的表达;透射电镜观察FB 超微结构的改变。结果 DM 大鼠烫伤前处于 G_0/G_1期细胞较低(DM 组66%±5%,正常组82%±10,P<0.01),烫伤后各时相点处于 G_0/G_1期细胞比例高于正常大鼠(烫伤后3 d,DM 组70%±4%,正常组42%±13%,P<0.01),S 期细胞低于正常大鼠(P<0.05,P<0.01);DM 大鼠皮肤组织单位面积 OHP 含量,烫伤前(DM 组0.72μg/cm^2±0.06μg/cm^2,正常组1.42μg/cm^2±0.28μg/cm^2,P<0.01)和烫伤后均低于正常大鼠(均 P<0.01),且伤后7、14和21d,创面Ⅰ/Ⅲ胶原比例明显高于正常组(烫伤后7 d,DM 组为5.54%±0.47%,正常组为4.01%±0.32%,P<0.01);DM 组α-SMA 的表达延迟,上升缓慢,且伤后各时相点的峰值均低于正常组;伤后 DM 组真皮层成纤维细胞的内质网扩张,高尔基复合体欠发达,微管微丝缺乏,且胞浆中溶酶体明显增多。结论 DM 创面修复延迟可能与 FB 增殖异常、胶原合成能力减弱、α-SMA 表达不足以及 FB 超微结构改变等有关。
Objective To explore the changes of the biological function of dermal fibroblasts （FBs） in the wounds of diabetic and non-diabetic burned rats and the pathogenesis of impaired wound healing in diabetes. Methods 80 Sprague-Dawley （SD） rats weighing 220 g were randomly divided into control and STZ-induced diabetic groups, and then deep partial thickness scald involving 10% TBSA was reproduced in the two groups. The diabetic groups were randomized into pre-scalding, post-scalding day（ PSD 3 ）, PSD 7, PSD 14 and PSD 21 groups, with 6 rats in each group. Controls were also randomized into 5 groups. Skin specimens from the wound were harvested at each time point. Cell cycles of FBs were analyzed with flow cytometry. The amount of hydroxyproline in the skin tissue was assessed on 0, 3, 7,14,and PSD 21. The type Ⅰ and III collagens were determined by ELISA. The expression of α-SMA in the dermal fibroblasts of each group was assessed by immunohistochemistry method. Transmission electron microscopy was used to observe the ultrastructure changes of FBs. Results Compared with that in the normal rats, the percentage of the ceils in G0/G, phase in the DM group was evidently lower on PSD 0 （65.79 ± 5.24 vs 82.43 ± 9. 68, P〈0. 01 ）. After the scalding, the percentage of the cells in G0/G1 phase in DM group was significantly higher（70. 00 ±4. 27 vs 42.04 ± 12.96, on PSD 3, P 〈0. 01 ）, meanwhile the percentage of S phase was remarkably lower than those in C group on 3, 7, 14,21PSD（P 〈0.05, P 〈0.01）.The amount of hydroxyproline in the diabetic skin tissue was obviously lower than those of the responding control groups before （0. 72 ±0.06 vs 1.42 ±0. 28, P 〈0. 01 ）and after burn injury （P 〈0. 01 ）. Furthermore, the rate of Ⅰ/Ⅲ collagen on 7, 14 and PSD 21 was much higher in DM group than that in C group （ P 〈0. 01 ）. The expression of α-SMA in DM groups on PSDS 3, 7, 14 and PSD 21 was evidently lower than those of the controls （ levels 10. 28 ± 3.99, C group 28.42 ±
出处
《中华医学杂志》
CAS
CSCD
北大核心
2007年第26期1812-1816,共5页
National Medical Journal of China
基金
国家重点基础研究发展规划基金(C1999054205)
关键词
糖尿病
烫伤
成纤维细胞
创面愈合
Diabetes mellitu
Burns
Dermal Fibroblast
Wound healing
