Zhu Xudong, Di Yanfei, Hu Chengxiang, Wang Zhengguo
Research Institute of Surgery, the Third Military Medical University, Chongqing 400042, China.
Chin Med J (Engl). 2002 Mar;115(3):326-30.
To detect the expression of cell cycle positive regulators cyclin D(1), cyclin E, CDK(2), CDK(4) and negative regulators p21(cip1), p27(kip1), p16(ink4a) and p15(ink4b) during wound healing in rats.
Open wounds of full-thickness skin, diameter 1.8 cm, on rat backs were used as the wound model. Wound tissues were harvested on postwounding days 3, 5, 7, 9, 11, 14, 21 and 30. Ki67 expression in granulation tissue was detected by immunohistochemical assay. The patterns of the expression of cyclin D(1), cyclin E, CDK(2), CDK(4) and p21(cip1), p27(kip1), p16(ink4a), p15(ink4b) were detected by Western blot.
Cell proliferation in granulation tissue took place predominantly within the first week after injury, with the proliferation peak occurring at postwounding day 5. There were no dramatic variations in the expression of cyclin D(1), CDK(2) and CDK(4) during wound healing. Up-regulated cyclin E was maintained from day 3 to 11 after injury, and then was down-regulated. No expression of p16(ink4a) and p15(ink4b) was found. p21(cip1) was expressed only from day 7 to 14, with peak expression observed on day 9. Constitutive p27(kip1) was expressed throughout wound healing with low levels in the proliferating period of day 3 to 5 and with increased levels in the post-mitotic and remodeling stage. The expression of p21(cip1) and p27(kip1) showed an inverse gradient to that of Ki67.
p21(cip1) and p27(kip1) play a supervising role in preventing the hyperproliferative tendency in tissue repair.
检测细胞周期正调控因子细胞周期蛋白D（1）、细胞周期蛋白E、细胞周期蛋白依赖性激酶（2）、细胞周期蛋白依赖性激酶（4）以及负调控因子p21（cip1）、p27（kip1）、p16（ink4a）和p15（ink4b）在大鼠伤口愈合过程中的表达情况。
以大鼠背部直径1.8 cm的全层皮肤开放性伤口作为伤口模型。在伤后第3、5、7、9、11、14、21和30天采集伤口组织。采用免疫组织化学法检测肉芽组织中Ki67的表达。采用蛋白质免疫印迹法检测细胞周期蛋白D（1）、细胞周期蛋白E、细胞周期蛋白依赖性激酶（2）、细胞周期蛋白依赖性激酶（4）以及p21（cip1）、p27（kip1）、p16（ink4a）、p15（ink4b）的表达模式。
肉芽组织中的细胞增殖主要发生在损伤后的第一周，增殖高峰出现在伤后第5天。在伤口愈合过程中，细胞周期蛋白D（1）、细胞周期蛋白依赖性激酶（2）和细胞周期蛋白依赖性激酶（4）的表达没有显著变化。伤后第3天至11天，细胞周期蛋白E表达上调，随后下调。未发现p16（ink4a）和p15（ink4b）的表达。p21（cip1）仅在第7天至14天表达，在第9天观察到表达高峰。组成型p27（kip1）在整个伤口愈合过程中均有表达，在第3天至5天的增殖期表达水平较低，在有丝分裂后和重塑阶段表达水平升高。p21（cip1）和p27（kip1）的表达与Ki67的表达呈相反梯度。
p21（cip1）和p27（kip1）在防止组织修复过程中的过度增殖倾向方面发挥监督作用。
