基于miR-128-3p/SIRT1/自噬探讨虎杖苷促进糖尿病溃疡模型大鼠创面愈合的机制  

To Explore the Mechanism of Polydatin Promoting Wound Healing in Diabetic Ulcer Model Rats Based on MiR-128-3p/SIRT1/Autophagy

作　　者：童海江[1] 周恺骅 王亚玲[1] 孙海燕[1] 王社梁[1] 夏伟仁[1] TONG Haijiang;ZHOU Kaihua;WANG Yaling;SUN Haiyan;WANG Sheliang;XIA Weiren(The Second Hospital of Shaoxing,Shaoxing Zhejiang 312000,China)

机构地区：[1]绍兴第二医院,浙江绍兴312000

出　　处：《中医药导报》2023年第8期13-18,共6页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：浙江省中医药科技计划中医药现代化专项项目(2021ZB309)。

摘　　要：目的:探究虎杖苷调控微小RNA(microRNA,miR)-128-3p/SIRT/自噬促进糖尿病(DM)溃疡模型大鼠创面愈合的机制。方法:通过腹腔注射链脲佐菌素(70 mg/kg)建立DM模型。将30只DM大鼠随机分为模型组(n=15)、虎杖苷组(n=15)。另取15只健康大鼠作为对照组。3组均通过去皮建立DM伤口损伤模型。虎杖苷组大鼠灌胃虎杖苷(20 mg/kg)。比较各组大鼠伤口愈合情况、愈合周围组织中超氧化物歧化酶(SOD)、丙二醛(MDA)和自噬标志蛋白表达水平。分析miR-128-3p和SIRT1的表达水平。通过双荧光素酶报告在内皮祖细胞中验证miR-128-3p与SIRT1的靶向关系。通过转染miR-128-3p mimic质粒过表达miR-128-3p。结果:第5天,模型组、虎杖苷组大鼠伤口愈合率低于对照组(P<0.05);第10天,虎杖苷组大鼠伤口愈合率高于模型组(P<0.05),低于对照组(P<0.05)。模型组大鼠创面组织中SOD、SIRT1 m RNA相对表达量、SIRT1蛋白相对表达量、LC3Ⅱ蛋白相对表达量、Beclin1蛋白相对表达量、LC3Ⅱ/Ⅰ均低于对照组(P<0.05),而MDA和miR-128-3p水平显著高于对照组(P<0.05)。虎杖苷组大鼠创面组织中SOD、SIRT1 m RNA相对表达量、SIRT1蛋白相对表达量、LC3Ⅱ蛋白相对表达量、Beclin1蛋白相对表达量、LC3Ⅱ/Ⅰ均高于模型组(P<0.05),而MDA、miR-128-3p均低于模型组(P<0.05)。双荧光素酶报告结果显示miR-128-3p可以与SIRT1靶向结合。转染miR-128-3p mimic后,内皮祖细胞中的miR-128-3p的水平显著升高,并且SIRT1 m RNA和SIRT1蛋白表达水平显著降低。结论:虎杖苷可能通过调控miR-128-3p/SIRT1通路诱导自噬,进而缓解高血糖引起的氧化应激损伤,促进DM模型大鼠的伤口愈合。Objective:To explore the mechanism of polydatin regulating miR-128-3p/SIRT/autophagy to promote wound healing in diabetic(DM)ulcer model rats.Methods:The DM model was established by intraperitoneal injection of streptozotocin(70 mg/kg).A total of 30 DM rats were randomly divided into model group and polydatin group,with 15 rats in each group.Another 15 healthy rats were selected as the control group.The DM wound injury model was established by peeling in the three groups.The rats in polydatin group were given polydatin(20 mg/kg)by gavage.The wound healing,the expression levels of superoxide dismutase(SOD),malondialdehyde(MDA)and autophagy marker proteins in the surrounding tissues of rats in each group were compared.The expression levels of miR-128-3p and SIRT1 were detected.The targeting relationship between miR-128-3p and SIRT1 was verified in endothelial progenitor cells by dual luciferase report.MiR-128-3p was overexpressed by transfection of miR-128-3p mimic plasmid.Results:On the 5th day,the wound healing rate of rats in the model group and polydatin group was lower than that in the control group(P<0.05).On the 10th day,the wound healing rate of rats in the polydatin group was higher than that in the model group(P<0.05),and lower than that in the control group(P<0.05).The relative expression levels of SOD,SIRT1 mRNA,SIRT1 protein,LC3Ⅱprotein,Beclin1 protein and LC3Ⅱ/Ⅰin the wound tissue of rats in the model group were lower than those in the control group(P<0.05),while the levels of MDA and miR-128-3 p were significantly higher than those in the control group(P<0.05).The relative expression of SOD,SIRT1 mRNA,SIRT1 protein,LC3Ⅱprotein,Beclin1 protein and LC3Ⅱ/Ⅰin wound tissue of rats in polydatin group were higher than those in model group(P<0.05),while MDA and miR-128-3 p were lower than those in model group(P<0.05).The results of dual luciferase report showed that miR-128-3p could bind to SIRT1.After transfection of miR-128-3p mimic,the level of miR-128-3p in endothelial progenitor cells was si

关 键 词：糖尿病足溃病 伤口愈合 虎杖苷 氧化应激 自噬 微小RNA-128-3p SIRT1 

分 类 号：R285.5[医药卫生—中药学]