糖尿病大鼠创面愈合过程中成纤维细胞增殖及Ⅰ型胶原合成减少

Decreased fibroblast proliferation and collagen content in the wound of diabetic rats caused by streptozotocin

目的:观察链脲佐菌素诱导的大鼠糖尿病复合创伤修复过程中成纤维细胞增殖与胶原合成的变化。方法:实验采用Wistar大鼠112只,随机分对照组和模型组。模型组大鼠腹腔注射链脲佐菌素(STZ)55mg/kg,3周后各组动物复合背部2.04cm2全厚皮切除形成伤口。观察创面愈合时间和愈合率;采用HE染色和免疫组化法观察成纤维细胞和增殖细胞核抗原(PCNA)表达水平;采用苦味酸-天狼星红染色和图像分析技术观察创面Ⅰ、Ⅲ型胶原含量及Ⅰ/Ⅲ型比值。结果:STZ诱发的糖尿病大鼠复合创伤后创面的愈合时间为(27.13±1.81)d,明显长于对照组(15.25±1.67)d,P<0.01;模型组在创伤第3、7和15d创面愈合率明显低于对照组,分别P<0.01;在3、5、7和9d模型组创面成纤维细胞数量和PCNA表达也明显少于对照组,分别P<0.05和P<0.01。两组创面在不同时点上Ⅰ型胶原分布均呈递增趋势,但对照组明显多于模型组,分别P<0.05;尽管对照组在创伤3、7dⅢ型胶原含量高于模型组,但在创伤3、7和11d模型组Ⅰ/Ⅲ胶原比值都明显低于对照组,分别P<0.01。结论:STZ可能通过影响创面细胞增殖和创面胶原合成,从而导致创面愈合迟缓。

AIM： To study fibroblast proliferation and collagen synthesis during wound healing in diabetic rats induced by streptozotocin. METHODS： 30 Wistar male rats were randomly divided into control group and model group. 55 mg/kg STZ were given intraperitoneally to model rats. After 3 weeks, a round skin of 2.04 cm^2 was excised on all dorsal back of rats. The healing time and healing rate were observed according to re - epithelization. The numbers of fibroblasts and the expression of proliferating cell nuclear antigen （PCNA） were observed by Hematoxylin- Eosin （HE） staining and immuno- histochemistry assay. Collagen Ⅰ and Ⅲ stained by Picric acid - Sirius red were calculated by image analysis. RESULTS： The healing time in model group was （27.13 ± 1.81 ） days, significantly longer than that in control group [ （ 15.25 ± 1.67） days, P 〈 0.01 ]. The healing rates in model group were significantly less than that in control group at day 3, day 7 and day 15 （P 〈 0.01）. The amount of fibroblasts and the expression of PCNA in model group were significantly less than those in control group on day 3, day 5, day 7 and day 9, respectively （P 〈 0,05, P 〈 0.01 ）. Even the content of collagen I in the wound of beth groups increased with time, the values were much higher than that in model group at different times （ P 〈 0.05）, respectively. For model group, the ratio of collagen Ⅰ/Ⅲ was less than that in control group 3, 7 and 11 days after wound （ P 〈 0.01 ）. CONCLUSION： STZ impaires wound healing in rats, which is possible caused by the disturbance of fihroblast proliferation and collagen synthesis in the wound.