低能量体外震波对糖尿病大鼠皮肤切割伤愈合强度的影响及其作用机制

Effect of low-energy extracorporeal shock wave on incisional wound breaking strength in diabetic rats and underlying mechanism

目的观察低能量体外震波(ESW)对糖尿病大鼠皮肤切割伤愈合强度的影响并探讨其在切口愈合过程中的作用机制。方法36只雄性SD大鼠(购自中国科学院实验动物中心)随机分为3组:空白对照组、糖尿组对照组和ESW治疗组,每组各12只。糖尿对照组和ESW治疗组使用链脲佐菌素法建立糖尿病大鼠模型。3周后于各组大鼠背部制作切割伤模型。ESW治疗组在伤后1、3、5 d行ESW治疗,糖尿病对照组和正常对照组仅涂抹耦合液,未行ESW治疗。伤后7 d和14 d收集标本,万能材料测试仪测量切口愈合强度,苏木精-伊红(HE)和Masson染色观察组织愈合,并检测切口组织内羟脯氨酸含量和转化生长因子-β1(TGF-β1)的表达。组间比较采用t检验和One-way ANOVA检测。结果在伤后7 d和14 d,ESW组切口的愈合强度明显高于糖尿病对照组,差异有统计学意义[(5.12±0.80)N比(2.38±0.67)N,F=1.602,t=-9.078,P<0.05;(12.80±2.49)N比(6.11±0.89)N,F=17.410,t=-7.289,P<0.05];ESW组羟脯氨酸含量明显高于糖尿病对照组[(15.66±1.89)mg/g比(8.60±1.79)mg/g,F=0.018,t=-6.639,P<0.05;(22.46±3.21)mg/g比(11.69±2.26)mg/g,F=0.082,t=-5.928,P<0.05];TGF-β1阳性表达主要位于切口的成纤维细胞内,并且在伤后7 d和14 d ESW治疗组TGF-β1蛋白表达明显高于糖尿病对照组,差异有统计学意义(0.42±0.05比0.14±0.02,F=2.901,t=-15.294,P<0.05;0.36±0.02比0.20±0.01,F=2.447,t=-14.482,P<0.05)。结论低能量ESW可以增加糖尿病皮肤切割伤的愈合强度,这与ESW上调成纤维细胞TGF-β1的表达,进而增加切口内的胶原合成有关。

Objective To investigate the effect of low-energy extracorporeal shock wave(ESW)on incisional wound breaking strength and its mechanism in diabetic rats.Methods Twenty-four male Sprague-Dawley rats were intraperitoneally injected with streptozotocin(STZ)to establish diabetic models and randomly divided into two groups:diabetic control group and ESW group(n=12 each).Twelve rats served as normal control group.In ESW group,rats received three sessions of ESW on days 1,3 and 5 post-wounding.Rats were sacrificed on days 7 and 14 post-wounding.Wound breaking strength,hydroxyproline content,histological evaluation and the expression of transforming growth factor beta 1(TGF-β1)were analyzed.Results On days 7 and 14 post-wounding,the wound breaking strength was significantly increased in ESW group as compared with that in diabetic control group[(5.12±0.80)N vs.(2.38±0.67)N,F=1.602,t=-9.078,P<0.05;(12.80±2.49)N vs.(6.11±0.89)N,F=17.410,t=-7.289,P<0.05].The hydroxyproline content was significantly increased in ESW group as compared with that in diabetic control group[(15.66±1.89)vs.(8.60±1.79)mg/g,F=0.018,t=-6.639,P<0.05;(22.46±3.21)vs.(11.69±2.26)mg/g,F=0.082,t=-5.928,P<0.05].The TGF-β1 expression was detected in fibroblasts,and the Western blotting showed that the expression of TGF-β1 was significantly increased in ESW group as compared with that in diabetic control group(0.42±0.05 vs.0.14±0.02,F=2.901,t=-15.294,P<0.05;0.36±0.02 vs.0.20±0.01,F=2.447,t=-14.482,P<0.05).Conclusion Low energy ESW can increase incisional wound breaking strength in diabetic rats probably by up-regulating the expression of TGF-β1 of fibroblasts in wound and then increasing the collagen synthesis.