咪喹莫特对兔耳瘢痕增生抑制作用的实验分析-experimental analysis of imiquimods inhibitory effect on rabbit ear scar hyperplasia.docxVIP

1 1 咪喹莫特对兔耳瘢痕模型增生抑制作用的 实验研究 中 文 摘 要 目的：研究局部外用免疫调节剂咪喹莫特乳膏对兔耳瘢痕增生是否存 在抑制作用及可能机制。 方法：选取 10 只新西兰大耳白兔，雌雄不限，根据 Morris 和李荟元 报道[1,2]及曾海峰[3]的方法建立兔耳瘢痕模型：每只兔耳腹侧做六个直径为 1cm 的圆形全层皮肤缺损创面，每个创面相距 1.5cm，双侧对称。待术后 14 天上皮化均完全后开始涂药，左耳为实验组涂抹 5% 咪喹莫特乳膏，右 耳为对照组涂抹等量凡士林软膏，一日一次。分别于术后第 14 天、21 天、 28 天、35 天及 42 天同一时间点空气栓塞处死 2 只兔子，后沿每个瘢痕边 缘外周 0.5cm 处的正常皮肤环形切下，并经瘢痕最凸点直径一切为二，一 半固定后行苏木精-伊红（Hematoxylin-eosin, HE）染色及马松（Masson） 三色法染色，观察形态学差异，测量并计算瘢痕增生指数（ Scar elevation index，SEI）；另一半用于提取 RNA，反转录后行 Real-time PCR 检测细胞 外基质Ⅰ型胶原（Collagen-Ⅰ, Col-Ⅰ），细胞因子 γ 干扰素（Interferon-γ, IFN-γ）及白细胞介素 4（Interleukin-4, IL-4） 的表达。 结果：HE 及 Masson 染色可见实验组胶原沉积较对照组明显减少， SEI 显示空白对照组于术后第 21 天增生程度达到高峰，其增生程度明显高 于实验组；Real-time PCR 结果可见实验组 Col-Ⅰ 及 IL-4 的表达在各时 间点明显降低，且 IFN-γ 的表达较对照组增高。 结论：咪喹莫特可能通过调节 IFN-γ 及 IL-4 的表达来发挥抑制瘢痕 增生的作用。 PAGE 1 PAGE 1 PAGE 2 关键词：咪喹莫特；兔耳瘢痕模型；T 淋巴细胞；增生性瘢痕 PAGE 1 PAGE 1 PAGE 3 Imiquimod inhibit scar formation in rabbit ear hypertrophic scar model ABSTRACT Objective: In this study, we evaluated the inhibitory effect and mechanism of imiquimod on rabbit ear hypertrophic scar formation. Methods:Ten New Zealand White rabbits were employed in this study. Rabbit ear hypertrophic scar model was created on the basis of the previous literatures reported by Morris, Huiquan Li and Haifeng Zeng. We cut six round wounds in each rabbit ears ventral that diameter of 1cm, bilateral symmetry, peel the full-thickness of skin and film layer of cartilage. All the left ear wounds were treated with 5% imiquimod cream. The control group contains all the right ear wounds and were treated with vaseline ointment at the same time. Every wounds were applied cream after healing, once a day .Both of the groups were harvested at the same time on days 14、21、28、35 and 42 after operation. Two rabbits were killed by air embolism every time. The specimens were cut along the edge outside the scar at 0.5cm ring, then cut through the most thickest point in half. HE (Hematoxylin-eosin) staining and Masson trichrome staining were performed after