中国修复重建外科杂志

中国修复重建外科杂志

曲尼司特对小鼠烧伤后创面愈合的影响及给药时间研究

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目的 探讨曲尼司特对小鼠烧伤创面愈合时间的影响及其抑制瘢痕增生的作用机制,以及曲尼司特抑制瘢痕增生的能力是否随给药时间点变化而变化。 方法 7~8 周龄清洁级雄性昆明小鼠 66 只,于小鼠背部制备深Ⅱ度烧伤模型,然后随机分为对照组(n=18)、早期干预组(n=18)、中期干预组(n=18)和晚期干预组(n=12)。早、中、晚期干预组分别于造模后当天、7 d、14 d 开始给予曲尼司特 200 mg/(kg·d)灌胃,对照组于造模后当天开始每天给予等体积生理盐水灌胃。定期观察创面愈合情况,对照组及早、中期干预组分别于造模后 14、28、42 d,晚期干预组于 28、42 d,随机选取 6 只小鼠取创面新生组织,分别行甲苯胺蓝染色观察肥大细胞形态,Masson 染色观察总胶原含量情况;行免疫组织化学染色观测Ⅰ型胶原及Ⅲ型胶原含量,并计算Ⅰ/Ⅲ型胶原含量比值;ELISA 法检测组织中 TGF-β1 和组胺含量;透射电镜观察成纤维细胞的超微结构。 结果 各组小鼠创面愈合时间比较差异无统计学意义(F=1.105,P=0.371)。早期干预组各时间点肥大细胞数、总胶原含量、Ⅰ/Ⅲ型胶原含量比值、TGF-β1 含量及组胺含量均显著低于其他组(P<0.05);除 42 d 时对照组肥大细胞数、总胶原含量、组胺含量与晚期干预组比较差异无统计学意义(P>0.05)外,其余各时间点对照组及中、晚期干预组间以上指标比较差异均有统计学意义(P<0.05)。与对照组相比,早期干预组成纤维细胞活跃程度明显被抑制,纤维排列更加规律整齐;中、晚期干预组成纤维细胞活性亦较明显被抑制。 结论 曲尼司特干预对小鼠烧伤创面愈合时间无明显影响,但能明显降低新生组织中肥大细胞、组胺和 TGF-β1 含量,抑制成纤维细胞胶原合成能力及调节胶原合成的比值,从而抑制瘢痕增生,且烧伤后即刻进行曲尼司特干预对瘢痕增生的抑制作用最明显。

Objective To investigate the effect of tranilast on wound healing and the mechanism of inhibiting scar hyperplasia in mice, and to study the relationship between the inhibiting ability of tranilast on scar hyperplasia and administration time. Methods Sixty-six Kunming mice were selected to build deep II degree burn model, and were randomly divided into the control group (18 mice), the early intervention group (18 mice), the medium intervention group (18 mice), and the late intervention group (12 mice). The mice in the early intervention group, the medium-term intervention group, and the late intervention group were given tranilast 200 mg/(kg·d) by gastrogavage at immediate, 7 days, and 14 days after burn respectively, and the mice in the control group were managed with same amount of normal saline every day. The wound healing was observed regularly. At 14, 28, and 42 days in the early and medium intervention groups and at 28 and 42 days in the late intervention group, fresh tissues were taken from 6 mice to observe the shape of mast cells by toluidine blue staining, collagen content by Masson staining; the collagen type I and collagen type III content were measured to calculate the I/III collagen content ratio by immunohistochemistry method, the contents of transforming growth factor β1 (TGF-β1) and histamine were detected by ELISA; and the ultrastructure of fibroblasts was observed under transmission electron microscope. Results There was no significant difference in wound healing time between groups (F=1.105,P=0.371). The mast cells number, collagen content, TGF-β1 content, histamine content, and the I/III collagen content ratio in the early intervention group were significantly less than those in the other groups (P<0.05). Significant difference was found in mast cells number, collagen content, and histamine content between control group and medium or late intervention group at the other time points (P<0.05) except between control group and late intervention group at 42 days (P>0.05). Compared with the control group, the activity of fibroblasts in the early intervention group was obviously inhibited, and the arrangement of the fibers was more regular; the fibroblast activity in the medium and late intervention groups was also inhibited obviously. Conclusion Tranilast has no obvious effect on the wound healing time in mice. Tranilast intervention shows the inhibitory effect on the scar hyperplasia which can significantly reduce the number of mast cells, the content of histamine and TGF-β1, inhibit the ability of fibroblasts synthetic collagen and adjust the proportion of collagen synthesis. The immediate tranilast intervention may have the best inhibitory effect on scar hyperplasia.

关键词: 曲尼司特; 瘢痕增生; 创面愈合; TGF-β1; 肥大细胞; 小鼠

Key words: Tranilast; scar hyperplasia; wound healing; transforming growth factor β1; mast cells; mouse

引用本文: 胡珍珍, 陈彬, 李阳, 姜卫, 文丽红, 姬付康, 杨晓, 王晋煌, 柳大烈. 曲尼司特对小鼠烧伤后创面愈合的影响及给药时间研究. 中国修复重建外科杂志, 2017, 31(4): 465-472. doi: 10.7507/1002-1892.201611033 复制

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