丹参联合罗沙司他对糖尿病大鼠全层皮肤缺损创面愈合的影响及其机制  

Effect of salvia miltiorrhiza combined with roxadustat on wound healing of full-thickness skin defects in diabetic rats and its mechanism

作　　者：夏如意 唐棣 杨斌 Xia Ruyi;Tang Di;Yang Bin(The First Clinical College of Hubei University of Chinese Medicine,Wuhan 430060,China;Department of Burns and Plastic Surgery,Central Theater Command General Hospital,Wuhan 430070,China)

机构地区：[1]湖北中医药大学第一临床学院,武汉430060 [2]中部战区总医院烧伤整形科,武汉430070

出　　处：《中华烧伤与创面修复杂志》2024年第4期380-388,共9页Chinese Journal of Burns And Wounds

基　　金：国家自然科学基金面上项目(81872536);国家自然科学基金青年科学基金项目(82302818);湖北省自然科学基金(2021CFB164)。

摘　　要：目的探究丹参联合罗沙司他对糖尿病大鼠全层皮肤缺损创面愈合的影响及其机制。方法该研究为实验研究。将20只8周龄雄性SD大鼠成功制成糖尿病模型后,在其背部制作全层皮肤缺损创面,之后将大鼠按照随机数字表法分为生理盐水组、单纯罗沙司他组、单纯丹参组、罗沙司他+丹参组,每组5只。伤后即刻,对生理盐水组大鼠予5 mL生理盐水灌胃,对单纯罗沙司他组大鼠予1.5 mg/mL的罗沙司他混悬液按照25 mg/kg灌胃,对单纯丹参组大鼠予18 mg/mL的丹参混悬液按照300 mg/kg灌胃,对罗沙司他+丹参组的大鼠予19.5 mg/mL的罗沙司他和丹参混悬液按照罗沙司他25 mg/kg、丹参300 mg/kg灌胃,均持续给药2周,1次/d。观察伤后0(即刻)、4、8、12 d创面情况,计算伤后4、8、12 d的创面愈合率(样本数为5)。伤后14 d,取腹主动脉血行血红蛋白、红细胞计数、白细胞计数检测(样本数为5);取创面组织,行苏木精-伊红染色后观察炎性浸润、皮肤组织结构及新生血管生成情况,行Masson染色后观测胶原纤维占比(样本数为3),行蛋白质印迹法检测血管内皮生长因子(VEGF)、CD31、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)、IL-1β的蛋白表达水平(样本数为3),行免疫组织化学染色检测表皮生长因子受体(EGFR)、缺氧诱导因子1α(HIF-1α)、增殖细胞核抗原(PCNA)的蛋白表达水平(样本数为3)。结果伤后0~12 d,4组大鼠创面面积均逐渐缩小。伤后4 d,单纯丹参组、罗沙司他+丹参组大鼠创面愈合率均明显高于生理盐水组和单纯罗沙司他组(P<0.05);伤后8 d,单纯罗沙司他组和单纯丹参组大鼠创面愈合率均明显高于生理盐水组(P<0.05),罗沙司他+丹参组大鼠创面愈合率明显高于其余3组(P值均<0.05);伤后12 d,单纯罗沙司他组、单纯丹参组、罗沙司他+丹参组大鼠创面愈合率均明显高于生理盐水组(P<0.05)。伤后14 d,4组大鼠的血红蛋白和红细胞Objective To explore the effect of salvia miltiorrhiza combined with roxadustat on wound healing of full-thickness skin defects in diabetic rats and its mechanism.Methods This study was an experimental study.Twenty male 8-week-old Sprague-Dawley rats were used to successfully establish diabetic model,then full-thickness skin defect wounds on their backs were made.The rats were divided into normal saline group,roxadustat alone group,salvia miltiorrhiza alone group,and roxadustat+salvia miltiorrhiza group according to the random number table,with 5 rats in each group.Immediately after injury,the rats in normal saline group were given 5 mL normal saline by gavage,the rats in roxadustat alone group were given 1.5 mg/mL roxadustat suspension by gavage at 25 mg/kg,the rats in salvia miltiorrhiza alone group were given 18 mg/mL salvia miltiorrhiza suspension by gavage at 300 mg/kg,and the rats in roxadustat+salvia miltiorrhiza group were given 19.5 mg/mL roxadustat and salvia miltiorrhiza suspension at roxadustat 25 mg/kg and salvia miltiorrhiza 300 mg/kg.All were administered once a day for 2 weeks.The wounds at 0(immediately),4,8,and 12 d after injury were observed,and the wound healing rates at 4,8,and 12 d after injury were calculated(n=5).At 14 d after injury,abdominal aortic blood was collected,and hemoglobin,red cell count,and white blood cell count were detected(n=5).The wound tissue was collected for hematoxylin-eosin staining to observe inflammatory infiltration,skin tissue structure,and neovascularization,for Masson staining to observe the proportion of collagen fiber(n=3),for Western blotting to detect the protein expression levels of vascular endothelial growth factor(VEGF),CD31,interleukin 6(IL-6),tumor necrosis factor α(TNF-α),and IL-1β(n=3),and for immunohistochemical staining to determine the protein expression levels of epidermal growth factor receptor(EGFR),hypoxia-inducible factor 1α(HIF-1α),and proliferating cell nuclear antigen(PCNA),with sample number of 3.Results From 0 to 12 d after injury,

关 键 词：糖尿病 实验性 伤口愈合 丹参 炎症 血管内皮生长因子类 受体 表皮生长因子 缺氧诱导因子1 Α亚基 罗沙司他 

分 类 号：R587.1[医药卫生—内分泌]