积雪苷软膏对BALB/C裸小鼠瘢痕疙瘩模型成纤维细胞及胶原蛋白表达的影响

Effect of asiaticoside ointment on fibroblast and collagen expression in the keloid model of nude mice

目的:明确外用积雪苷软膏对瘢痕疙瘩裸鼠模型的成纤维细胞和胶原蛋白表达的影响。方法:取手术切除的瘢痕疙瘩组织植入16只裸鼠肩胛及骶尾部,建立瘢痕疙瘩裸鼠模型。随机分为A组(外用对照软膏基质),B组(外用0.00125%积雪苷软膏),C组(外用0.0025%积雪苷软膏)和D组(外用0.005%积雪苷软膏),每组4只。观察瘢痕疙瘩大体形态学及光镜下变化;免疫组化检测瘢痕疙瘩I型胶原蛋白表达的影响;透射电镜观察瘢痕疙瘩成纤维细胞结构的变化。结果:大体观察裸鼠瘢痕疙瘩经不同浓度积雪苷软膏作用后组织块体积均变小,光镜观察成纤维细胞数明显减少。A^D组I型胶原蛋白的表达率分别为79.17%、70.83%、37.50%、33.33%,四组比较,差异具有统计学意义(Fisher精确概率检验法,P<0.05)。透射电镜观察B-D组瘢痕疙瘩成纤维细胞较A组明显减少,可见凋亡细胞,细胞核固缩,膜结构不清,有坏死的细胞碎片及裸核状态的成纤维细胞。结论:外用积雪苷软膏能减少瘢痕疙瘩成纤维细胞并下调I型胶原蛋白的表达。

Objective:To determine the effect of asiaticoside ointment on fibroblast and collagen expression in the keloid model of nude mice. Methods:Keloid tissue was removed and implanted into scapular and sacral tail of 16 nude mice, which were divided into four groups: group A (control group), group B (topical 0.00125% of asiaticoside), group C (topical 0.0025% of asiaticoside), group D (topical 0.005 of asiaticoside), four mice in each group. The change of keloid gross morphology was observed. The expression level of type I collagen in keloid was detected by immunohistochemistry. The changes of fibroblast structure in keloid were observed by projection electron microscopy. Results:Compared to group A, the size of keloid and the number of fibroblasts were decreased in group B-D. The expression rate of collagen in the experimental group A-D was 79.17%, 70.83%, 37.50% and 33.33%, with a significant difference among four groups (Fisher test, P <0.05). Compared to group A, the number of keloid fibroblasts in group B-D was significantly reduced and there were apoptotic cells, retracted nuclei, unclear cell membrane structure, necrotic cell fragments and fibroblasts in the state of naked nucleus under transmission electron microscopy. Conclusion:Topical application of asiaticoside can reduce keloid fibroblasts and down-regulate type I collagen expression.