丹黄消炎液对糖尿病小鼠创面组织中HIF1-α及VEGF表达的影响  

Effect of Danhuang Xiaoyan Solution on Expression of HIF1-α and VEGF in Wound Healing of Diabetic Mice

作　　者：刘国涛 王军[1] 周玉 卢增珍 LIU Guo-tao;WANG Jun;ZHOU Yu(Department of Surgery,the First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300381,China)

机构地区：[1]天津中医药大学第一附属医院疮疡周围血管外科

出　　处：《中国中西医结合外科杂志》2020年第1期20-26,共7页Chinese Journal of Surgery of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金(81973854)

摘　　要：目的:应用糖尿病小鼠模型,探讨丹黄消炎液外敷对糖尿病创面的治疗作用及相关机制。方法:通过多次小剂量腹腔注射链脲佐菌素(STZ)制作I型糖尿病小鼠模型,并切除小鼠15 mm×15 mm背部全层皮肤制作创面;将造模成功的小鼠随机分为模型组、阳性药组、中药组,每组10只;从普通小鼠创面模型中随机挑选10只作为正常组。从造模后第3天起,正常组和模型组用生理盐水1 mL/只外敷,阳性药组用碘伏1 mL/只外敷,中药组用丹黄消炎液1 mL/只外敷,医用自粘绷带包扎1次/d,连续外敷7 d。给药后观察并计算各组小鼠愈合面积,同时收集各组小鼠创面组织进行HE染色及Masson染色,检测各组小鼠创面中病理学变化及纤维组织分布,评价丹黄消炎液外敷对糖尿病小鼠创面的治疗作用;荧光定量PCR法检测各组小鼠创面组织中低氧诱导因子1-α(HIF1-α)及血管内皮生长因子(VEGF)mRNA表达水平,免疫组化法检测各组小鼠创面组织中HIF1-α、VEGF及CD31蛋白表达水平。结果:正常组、阳性药组、中药组创面愈合面积分别为(117.40±8.04)mm2、(115.76±14.34)mm2、(147.82±6.31)mm2,显著高于模型组(P<0.05);中药组愈合面积显著高于阳性药组(P<0.05);与模型组相比,正常组、阳性药组、中药组HIF1-αmRNA和VEGF mRNA表达水平显著上调(P<0.05),中药组HIF1-αmRNA表达水平与阳性药组相比显著上调(P<0.05);免疫组化结果显示,与模型组相比,正常组、阳性药组、中药组小鼠创面组织中CD31显著增加,且中药组高于阳性药组,差异有统计学意义(P<0.05)。HIF1-α及VEGF主要表达于纤维组织与血管内皮细胞的细胞质中。与模型组相比,正常组、阳性药组及中药组创面组织中HIF1-α及VEGF阳性表达水平显著增加(P<0.05)。中药组的HIF1-α与阳性药组相比无统计学差异,但两组HIF1-α蛋白阳性表达水平均低于正常组(P<0.05);中药组创面的VEGF蛋白表达水平显著高于�Objective To investigate the therapeutic effect and mechanisms of Danhuang Xiaoyan Solution(DHXY)in wound healing of diabetic mice.Methods Type I diabetic model mice were induced through intraperitoneal injection of streptozotocin(STZ).Then,the whole layer of skin on the back of mice was excised for 15 mm×15 mm to establish the wound model in type 1 diabetic mice.The mice were randomly divided into model group,positive control group and Chinese medicine group.Normal mice without STZ administration were used as control group.Mice in control and model groups received external application of normal saline whereas mice in positive control and Chinese medicine groups received external application of Iiodine and DHXY once daily for 7 days.The effects of DHXY on diabetic mice with wound healing were studied via calculating the area of wound.HE staining and Masson staining were also conducted to observe the pathological changes after DHXY treatment.In addition,PCR and immunostaining were conducted to investigate the gene and protein expression of HIF1-αand VEGF after DHXY treatment.Results DHXY treatment significantly reduced the area of wound,inhibited the infiltration of inflammatory cells and increased the fibers in wound.PCR and immunostaining indicated that DHXY increased the expression of HIF1-αand VEGF in wound.The results of immunohistochemistry showed that compared with the model group,CD31 was significantly increased in the wound tissues of mice in the normal group,the positive drug group and Chinese medicine group,and the difference was statistically significant(P<0.05).HIF1-αand VEGF are mainly expressed in the cytoplasm of fibrous tissue and vascular endothelial cells.Compared with the model group,the expression of HIF1-αand VEGF in the wound tissues of the normal group,the positive drug group and Chinese medicine group increased significantly(P<0.05).There was no significant difference between the Chinese medicine group and the positive control group,but the expression level of HIF1-αprotein in the tw

关 键 词：丹黄消炎液 创面愈合 缺氧诱导因子1-Α 血管内皮生长因子 

分 类 号：Q95-33[生物学—动物学] R285.5[医药卫生—中药学]