【作者】 李莉；

【导师】 廖立新；

【作者基本信息】 南昌大学 ， 外科学， 2010， 硕士

【摘要】 目的:增生性瘢痕严重影响患者的美观及功能,但它的防治仍是烧伤整形外科学领域的重要难题。本研究通过观察羟基喜树碱对成纤维细胞胶原蛋白合成及细胞信号转导抑制子Smad7表达影响,以期了解羟基喜树碱抑制瘢痕作用机制。方法:人增生性瘢痕标本取自本院整形患者,采用组织块法体外培养人增生性瘢痕成纤维细胞。(1)选取羟基喜树碱63,125,250,500,1000ng/ml共五个浓度梯度,采用ELISA法分别在24、48、72小时后检测细胞上清液中Ⅰ、Ⅲ型胶原蛋白量。(2) 125、250、500ng/ml浓度的羟基喜树碱作用瘢痕细胞24h后,采用逆转录-聚合酶链反应(RT-PCR)及蛋白印迹法(Western blot)分别观察Smad7 mRNA及蛋白的表达。结果:(1)与对照组相比,实验组各浓度(63,125,250,500,1000ng/ml)羟基喜树碱均抑制增生性瘢痕成纤维细胞Ⅰ、Ⅲ型胶原蛋白的分泌,且随着药物浓度增加,抑制作用增强,与作用时间无明显相关。(2)实验组各浓度HCPT作用24小时后Smad7mRNA表达分别为0.350±0.037、0.431±0.054、0.616±0.038,与对照组(0.215±0.020)比较,差异有统计学意义(P<0.05)。Smad7蛋白表达分别为0.431±0.027、0.540±0.043、1.186±0.148,与对照组(0.280±0.033)比较,差异有统计学意义(P<0.05)。这说明,各浓度HCPT干预后增生性瘢痕成纤维细胞Smad7mRNA及蛋白表达均增强。其中,500ng/mL浓度组促表达作用最强。结论:羟基喜树碱可通过抑制人增生性瘢痕成纤维细胞胶原蛋白的合成及促进瘢痕成纤维细胞Smad7的表达,来发挥抑制瘢痕形成的作用。更多还原

【Abstract】 Goal: Hypertrophic scar (HS) is severely affected people,s appearance and function, but prevention and treatment of HS is still a difficult problem in the area of burn and plastic surgery medical research. In order to understand the mechanism of HCPT on treat scar, this study observed the effects of 10-hydroxycamptothecin (HCPT) on fibroblast collagen synthesis and the expression of cell signal transduction inhibitor Smad7,Methods: HS obtained from patients who received plastic surgery, and then hypertrophic scar fibroblasts (HSFB) were cultured in vitro using tissue culture method. (1) HSFB was intervened by HCPT with various concentrations (63、125、250、500、1000ng/ml), the secretion of typeⅠ,Ⅲcollagen from HSFB were assayed with ELISA method after 24、48、72 hours. (2) HSFB was intervened by HCPT with various concentrations (125、250、500ng/ml) for 24 hours, then the expression of Smad7 mRNA was measured with RT-PCR and Smad7 protein was detected with western blot method.Results : (1) Compared with the control group, HCPT with various concentrations (63、125、250、500、1000ng/ml) can inhibit the secreation of typeⅠ,Ⅲcollagen, and as the concentration increased, the effect of inhibition raised. But there is no significant relation with action time. (2) Compared with control group (0.215±0.020), the expression of Smad7mRNA in experiment groups is 0.350±0.037、0.431±0.054、0.616±0.038. There is significant difference between control group and experiment group (P<0.05). the expression of Smad7 protein in experiment groups is 0.431±0.027、0.540±0.043、1.186±0.148. And there is significant difference compared with control group (0.280±0.033). (P<0.05). These is indicated that hydroxycamptothecin could significantly increase Smad7 protein and mRNA expression in human hypertrophic scar fibroblasts,especially when its concentration at 500ng/mL.Conclusion:Hydroxycamptothecin could inhibit collagen synthesis of HSFB, and enhanced the expression of Smad7 mRNA and protein, ,which has a role in the treatment of hypertrophic scars.更多还原