唐肌康对糖尿病大鼠难愈性溃疡创面成纤维细胞、Ⅰ、Ⅲ型胶原合成的影响

Effect of Tangjikang on fibroblasts and collagen Ⅰ,Ⅲ in intractable ulcer of diabetic rats

目的:建立大鼠糖尿病溃疡模型,对比研究唐肌康治疗难愈性溃疡时的疗效及探索机制。方法:30只大鼠用STZ35mg/kg造模8周,冰醋酸涂抹创面3 d,建立糖尿病难愈溃疡模型,分为2组,自身同体对照,左侧为唐肌康治疗侧,右侧分别为表皮生长因子和生理盐水治疗侧,分别观察溃疡创面愈合率并于7、14d分2批处死糖尿病大鼠,取其溃疡及周围皮肤组织,采用HE染色观察创而及其周围皮肤成纤维细胞数、新生血管数,免疫组化法观察Ⅰ、Ⅲ型胶原含量。结果:成功建立糖尿病难愈性溃疡模型,大鼠体重下降,差异有统计学意义(P<0.05);血糖升高均稳定大于20.0mmol/L,前后自身对比有统计学差异(P<0.05)。唐肌康侧愈合率于7d时高于表皮生长因子(rhEGF)(P<0.05),于5、7、11、14d时唐肌康侧愈合率高于生理盐水侧(P<0.05)。成纤维细胞于第7、14 d唐肌康侧均明显多于表皮生长因子侧和生理盐水侧(P<0.05)。唐肌康侧新生血管数于第7d时多于表皮生长因子侧,二者比较差异无统计学意义(P<0.05)。第14d时,唐肌康侧Ⅰ型胶原含最明显低于生理盐水侧,二者比较差异有统计学意义(P>0.05)。第14d时,Ⅲ型胶原含量唐肌康侧均高于表皮生长因子和生理盐水侧(P<0.05),唐肌康Ⅰ/Ⅲ型胶原比值总体呈下降趋势,唐肌康侧Ⅰ/Ⅲ胶原比值均低于生理盐水侧(P<0.05)。结论:唐肌康治疗糖尿病大鼠皮肤难愈性溃疡愈合率比生理盐水侧高,瘢痕少可能与唐肌康使成纤维细胞增多,新生血管增多,调节Ⅰ/Ⅲ胶原比值有关。

Objective：To establish intractactable skin ulcer model of diabetic rats and to study the therapeutic effect of Tangjikang and its mechanism. Methods：Intractable ulcer models were made by surgery and daub of glacial acetic acid on 30 STZ rats. Each rat had two symmetrical wounds on right and left side of back. Then Tangjikang was applied on left side and rhEGF or saline treatment on right ride. The wound healing rate was obserred, and the rats were killed at 7d and 14d, the wound and the surrounding skin tissue of the ulcer were taken, using HE staining to evaluate the count of fibroblasts and the new blood vessels and immuonhistochemstry to observe the expression of collagenⅠ, Ⅲ. Results：The healing rate of Tangjikang was higher than that of rh-EGF at 7d and saline at 5,7,11, 14d（P〈0.05）. The count of fibroblasts of Tangjikang side was more than that of rhEGF and saline side at 7,14d （P〈0.05）.The count of new blood vessels of Tangjikang side more than that of rhEGF side at 7day （P〈0.05）. At 7d, the collagen Ⅰwas more in Tangjikan side but at 14d was less than in saline side （P 〈 0.05）. The collagen Ⅲ of Tangjikang side was higher than rhEGF, and saline side,The of ratio Ⅰ/Ⅲ collagen was declining in Tangjikang side,at 14d it was lower than rhEGF and saline side （P〈0.05）. Conclusion：Tangjikang has significant effect. The healing rate of Tangjikang side is higher than that of saline side ,and the scar is less.It is likely due to the proliferation of fibroblasts,the increase of new blood vessels, and the regulation of the ratio of collagenⅠ/Ⅲ.