白芷通过缺氧诱导因子-1α介导血小板衍生的生长因子受体-β的表达促进糖尿病小鼠足创面愈合的实验研究  

Angelica dahurica via hypoxia inducible factor-1αmediates platelet derived growth factor receptors-βsecretion to promote wound healing in type 2 diabetic mice

作　　者：郭俊[1] 周玉[1] 雷思思 孙蓓[1] 潘从清[1] Guo Jun;Zhou Yu;Lei Sisi;Sun Bei;Pan Congqing(Department of Integrated Traditional Chinese and Western Medicine,National Health Commission Key Laboratory of Hormones and Development(Tianjin Medical University),Tianjin Key Laboratory of Metabolic Diseases,Tianjin Medical University Chu Hsien-I Memorial Hospital and Metabolic Diseases Hospital,Tianjin Medical University,Tianjin 300134,China)

机构地区：[1]天津医科大学朱宪彝纪念医院中西医结合科、天津市内分泌研究所、国家卫健委激素与发育重点实验室、天津市代谢性疾病重点实验室,天津300134

出　　处：《中华糖尿病杂志》2024年第10期1132-1140,共9页CHINESE JOURNAL OF DIABETES MELLITUS

基　　金：天津市教委科研计划项目(2020KJ190)。

摘　　要：目的探讨白芷是否通过缺氧诱导因子-1α(HIF-1α)/血小板衍生的生长因子受体-β(PDGFR-β)信号通路促进2型糖尿病小鼠血管生成和加速创面愈合。方法将48只自发2型糖尿病db/db雄性小鼠按照随机数字表法分为糖尿病溃疡组(db/db组)和白芷组(db/db+Angelica dahurica组),每组24只,另选取24只同龄同源db/m雄性小鼠作为正常溃疡对照组(db/m组)。采用皮肤打孔器在各组小鼠背部打孔制备皮肤溃疡模型。白芷组给予白芷颗粒1.8 g/kg灌胃,正常溃疡对照组和糖尿病溃疡组给予等量生理盐水灌胃。每天给药一次,连续给药11 d,分别于创伤后第1、4、7、11天处死小鼠取材分析,每次每组6只小鼠。使用HE、Masson染色观察创面血管形成及胶原纤维形成等组织形态学改变;免疫组织化学染色、免疫荧光双染及Western blotting法从蛋白水平检测白芷干预后db/db小鼠创面组织中血管化及新生血管成熟相关因子[磷酸化磷脂酰肌醇-3激酶/磷脂酰肌醇-3激酶(p-PI3K/PI3K)、磷酸化蛋白激酶B/蛋白激酶B(p-AKT/AKT)、HIF-1α和PDGFR-β]的表达情况。将人脑微血管周细胞(HBVP)分为以下5组:对照组、高糖组、白芷干预组、高渗对照组和白芷+PI3K抑制剂(LY294002)干预组,应用Western blotting法检测各组细胞中HIF-1α、p-PI3K/AKT、PDGFR-β的表达,使用PI3K抑制剂(LY294002)以及使用HIF-1α-小干扰RNA(siRNA)静默HIF-1α或pReceiver-M11-HIF-1α过表达HIF-1α后检测HBVP募集因子PDGFR-β的表达情况。组间比较采用单因素方差分析。结果动物实验显示,糖尿病溃疡组小鼠各时间点创面愈合率均明显低于正常溃疡对照组(P<0.05),创伤后第4、7、11天,白芷组的创面修复、缩短创面愈合时间均优于糖尿病溃疡组(P<0.05);与正常溃疡对照组比较,糖尿病溃疡组创面的HIF-1α和PDGFR-β阳性表达明显减少(P<0.05);与糖尿病溃疡组比较,白芷干预组上述因子的阳性表达明显增加(P<0.0Objective To determine whether the hypoxia inducible factor-1α(HIF-1α)/platelet-derived growth factor receptor-β(PDGFR-β)signaling pathway is the mechanism by which Angelica dahurica stimulates angiogenesis and promotes angiogenesis and accelerates wound healing in type 2 diabetic mice.Methods A total of 48 spontaneous type 2 diabetic db/db male mice were randomly assigned to two groups using the random number table method:diabetic ulcer group(db/db group)and Angelica dahurica group(db/db+Angelica dahurica),with 24 mice in each group,24 homologous db/m male mice of the same age were selected as the normal ulcer control group(db/m group).The skin ulcer model was prepared using a skin punch on the back of mice.The normal ulcer control group and the diabetes ulcer group received the same amount of normal saline by gavage,while the Angelica dahurica group received 1.8 g/kg of Angelica dahurica granules.The treatment was administered once daily for 11 consecutive days and the mice were euthanized for analysis at 1,4,7,and 11 days post-trauma.There were six mice in each group.HE and Masson staining were used to observe the histomorphologic changes of wound angiogenesis and collagen fiber formation.Western blotting,immunohistochemical staining,and immunofluorescence double staining were used to identify the expression of factors related to angiogenesis and neovascularization,such as phosphorylated phosphatidylinositol 3-kinase/phosphatidylinositol 3-kinase(p-PI3K/PI3K),phosphorylated protein kinase B/protein kinase B(p-AKT/AKT),HIF-1α,and PDGFR-β,in the wound tissues of db/db mice following Angelica dahurica intervention at the protein levels.Five groups were created from human brain microvascular pericytes(HBVP):control group,high glucose group,Angelica dahurica intervention group,high osmotic control group,and angelica dahurica+PI3K inhibitor(LY294002)intervention group.Before detecting the expression of the pericytes recruitment factor PDGFR-β,HIF-1αor pReceiver-M11-HIF-1αoverexpression was silenced using PI

关 键 词：糖尿病 伤口愈合 缺氧诱导因子1 Α亚基 白芷 血小板衍生生长因子受体-β 

分 类 号：R285.5[医药卫生—中药学]