鸡血藤对db/db糖尿病皮肤溃疡小鼠创面愈合的影响及作用机制研究  

Effects and Mechanism of Spatholobi Caulis on Wound Healing in db/db Mice with Diabetic Cutaneous Ulcers

作　　者：杨丹阳 张暇蔚 姚文涛 马慧可 陈佳 王亚卓 赵宁 朱昊玥 李琳[1] 李萍[1] YANG Danyang;ZHANG Xiawei;YAO Wentao;MA Huike;CHEN Jia;WANG Yazhuo;ZHAO Ning;ZHU Haoyue;LI Lin;LI Ping(Beijing Institute of Traditional Chinese Medicine,Beijing Hospital of Traditional Chinese Medicine of Capital Medical University,Beijing 100010,China)

机构地区：[1]首都医科大学附属北京中医医院,北京市中医药研究所,北京100010

出　　处：《世界中医药》2024年第23期3583-3591,共9页World Chinese Medicine

基　　金：国家自然科学基金项目(82204920);北京市自然科学基金项目(7242215);北京市财政专项(YJS-2023-01)。

摘　　要：目的:探索鸡血藤对db/db小鼠糖尿病皮肤溃疡(DCU)创面愈合的影响及其作用机制。方法:将10只db/m小鼠作为空白组;将db/db小鼠制备DCU模型并使用随机数字表法随机分为模型组、阳性对照组、鸡血藤组,每组10只。第0天起,空白组和模型组给予蒸馏水10 mL/kg灌胃,外用含50μL生理盐水明胶海绵;阳性对照组给予蒸馏水10 mL/kg灌胃,外用含50μL bFGF明胶海绵;鸡血藤组给予鸡血藤水提物10 mL/kg(即给药量3.86 g/kg·d)灌胃,外用含50μL生理盐水明胶海绵。每天灌胃1次,背部换药隔天1次,并拍摄创面照片。第21天,小鼠安乐死,取伤口组织。测量小鼠体质量、血糖,计算创面愈合率,经苏木精-伊红(HE)染色、马松(Masson)染色观察创面组织病理学变化,通过网络药理学结合分子对接预测鸡血藤治疗DCU的潜在通路及靶点。用微量法分别测定血清氧化应激指标(MDA、SOD)。实时荧光定量聚合酶链式反应(RT-qPCR)法检测创面组织中HIF-1α、VEGFA mRNA表达,免疫组织化学染色法、蛋白质免疫印迹法检测创面组织中HIF-1α、VEGFA蛋白表达。结果:干预第0、21天,与空白组比较,模型组小鼠体质量、血糖高(P<0.05);模型组、阳性对照组、鸡血藤组小鼠体质量、血糖比较差异无统计学意义(P>0.05)。干预第8、14、21天,与空白组比较,模型组创面愈合率低(P<0.05);与模型组比较,阳性对照组、鸡血藤组创面愈合率高(P<0.05)。与模型组比较,阳性对照组、鸡血藤组创面组织病理学善。筛选出17个鸡血藤抗DCU关键成分,68个“药物-疾病”共同作用靶点,核心靶点包括HIF-1α、VEGFA等,分子对接显示结合能绝对值最高为HIF-1α-芦荟大黄素。与空白组比较,模型组小鼠血清MDA水平高(P<0.05),SOD水平低(P<0.05)。与模型组比较,阳性对照组小鼠血清MDA、SOD水平差异无统计学意义(P>0.05);鸡血藤组MDA水平低(P<0.05),SOD水平高(P<0.05)。与空白组比较,模�Objective:To explore the effects of Spatholobi Caulis on wound healing in diabetic cutaneous ulcers(DCU)in db/db mice and its underlying mechanism.Methods:Ten db/m mice were assigned to the blank control group.The db/db mice were used to establish a DCU model and randomly divided into the model group,positive control group,and Spatholobi Caulis group according to the random number table method,with 10 mice per group.From day 0,the blank and model groups were given distilled water(10 mL/kg)by gavage,with a gelatin sponge soaked in 50μL of normal saline applied to the wounds.The positive control group received distilled water(10 mL/kg)by gavage,with a gelatin sponge soaked in 50μL of bFGF applied to the wounds.The Spatholobi Caulis group received Spatholobi Caulis water extract(10 mL/kg,corresponding to 3.86 g/kg·d)by gavage,with a gelatin sponge soaked in 50μL of normal saline applied to the wounds.Gavage was performed once daily,and wound dressing changes were done every other day,with wound photos taken.On day 21,the mice were euthanized,and the wound tissues were collected.Body weight,blood glucose,and wound healing rate were measured.Hematoxylin-eosin(HE)staining and Masson′s trichrome staining were performed to observe the histopathological changes in the wound tissue.Network pharmacology and molecular docking were used to predict the potential pathways and targets of Spatholobi Caulis in treating DCU.Serum oxidative stress indicators(MDA and SOD)were measured using a microplate method.RT-qPCR was used to detect the mRNA expression of HIF-1αand VEGFA in the wound tissue.Immunohistochemistry and Western blot were used to detect the protein expression of HIF-1αand VEGFA in the wound tissue.Results:On days 0 and 21,compared to the blank group,the model group showed higher body weight and blood glucose(P<0.05).No significant difference was found between the model,positive control,and Spatholobi Caulis groups for body weight and blood glucose(P>0.05).On days 8,14,and 21,the model group had a lower wound he

关 键 词：鸡血藤 糖尿病皮肤溃疡 创面愈合 氧化应激 缺氧诱导因子-1α/血管内皮生长因子信号通路 网络药理学 分子对接 小鼠 

分 类 号：R255.4[医药卫生—中医内科学] R282[医药卫生—中医学] R285