温阳生肌膏调控GRP78/CHOP通路抑制过度内质网应激促糖尿病难愈合创面修复  

Wenyang-Shengji ointment regulates GRP78/CHOP pathway to inhibit excessive endoplasmic reticulum stress for facilitating diabetic refractory wound repair

作　　者：丁雅容 郑时旭 王君[3] 谢晨磊 奉水华[1] 周忠志[1,2] 陈丽[2] DING Yarong;ZHENG Shixu;WANG Jun;XIE Chenlei;FENG Shuihua;ZHOU Zhongzhi;CHEN Li(Department of Burns,Ulcers and Plastic Surgery,The First Hospital of Hunan University of Chinese Medicine,Changsha 410007,China;Hunan Provincial Key Laboratory of Vascular Biology and Translational Medicine,Hunan University of Chinese Medicine,Changsha 410208,China;The People'S Hospital of Hainan Province,Haikou 570311,China)

机构地区：[1]湖南中医药大学第一附属医院烧伤疮疡整形科,湖南长沙410007 [2]湖南中医药大学血管生物学与转化医学湖南省重点实验室,湖南长沙410208 [3]海南省人民医院,海南海口570311

出　　处：《中国病理生理杂志》2024年第3期493-501,共9页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.82074447);海南省重点研发项目(No.ZDYF2023SHFZ123);湖南省自然科学基金资助项目(No.2023JJ60348);湖南省卫生健康委卫生科研课题(No.23A0294,No.D202304039377);湖南中医药大学省级学科建设项目(中西医结合);血管生物学与转化医学湖南省重点实验室(No.2023TP1018)

摘　　要：目的:通过分析葡萄糖调节蛋白78(CRP78)/CCAAT/增强子结合蛋白同源蛋白(CHOP)通路研究温阳生肌膏对内质网应激(ERS)的调控,以探讨温阳生肌膏促进糖尿病难愈合创面修复的相关机制。方法:SD大鼠高脂饲料喂养联合链脲霉素腹腔注射后手术制备背部全层皮肤缺损建立大鼠糖尿病难愈合创面模型;实验分为正常组、模型组、温阳生肌膏组和贝复新(重组牛碱性成纤维细胞生长因子外用凝胶)组;正常组及模型组消毒后予以生理盐水涂抹,温阳生肌膏组及贝复新组分别以温阳生肌膏、贝复新涂抹创面,每日换药1次;处理14 d后观察创面愈合情况,计算创面愈合率;HE染色观察创面组织显微形态;Western blot法检测创面组织GRP78、CHOP和cas pase-12含量;免疫组化法检测创面GRP78、CHOP和caspase-12表达及分布情况;透射电子显微镜观察创面内质网数量及肿胀情况;ELISA检测创面促炎因子白细胞介素1β(IL-1β)含量。结果:各组相应干预14 d后检测各项指标。与正常组相比,模型组创面愈合率显著降低(P<0.01),炎症渗出较多,肉芽组织生长情况差,GRP78、CHOP和caspase-12蛋白表达升高(P<0.01),炎症因子IL-1β含量显著增加(P<0.01);与模型组相比,温阳生肌膏组创面愈合率显著增加(P<0.01),炎症渗出减少,肉芽组织生长良好,创面组织GRP78、CHOP和caspase-12蛋白表达显著降低(P<0.01);炎症因子IL-1β含量显著降低(P<0.01)。结论:温阳生肌膏促进糖尿病难愈合创面愈合的作用,可能与其下调GRP78/CHOP通路,抑制过度ERS,降低创面细胞凋亡水平有关。AIM:To study the regulation of endoplasmic reticulum stress(ERS)using the glucose regulated protein 78(GRP78)/CCAAT/enhancer binding protein homologous protein(CHOP)pathway and explore the related mech anism of Wenyang-Shengji ointment in facilitating the repair of diabetic refractory wounds.METHODS:To establish a rat model of diabetic refractory wound repair,Sprague-Dawley(SD)rats were fed a high-fat diet and intraperitoneally in jected with streptozotocin.Subsequently,full-thickness skin defects were induced in the dorsal region of the rats.The ex periment included 4 groups:normal,model(diabetic refractory wounds),Wenyang-Shengji ointment,and Beifuxin(re combinant bovine basic fibroblast growth factor gel)groups.The normal and model groups were treated with normal saline after disinfection.In the Wenyang-Shengji ointment and Beifuxin groups,the wounds were topically treated with the re spective ointments once daily.After 14 d of treatment,wound healing was assessed and quantified using the wound healing rate.Hematoxylin-eosin(HE)staining was employed to examine the micromorphology of the wound tissue.Western blot analysis was performed to measure GRP78,CHOP and caspase-12 levels in the wound tissue.Immunohistochemical analy sis was used to detect the expression and distribution patterns of GRP78,CHOP and caspase-12 in the wounds.Transmis sion electron microscopy was used to observe reticulum numbers and swelling.Enzyme-linked immunosorbent assay was used to determine interleukin-1β(IL-1β)level as a pro-inflammatory factor within the wound.RESULTS:Indexes of each group were assessed 14 d after the corresponding intervention.Compared with normal group,the rats in model group exhibited a significant decrease in the wound healing rate(P<0.01),accompanied by increased inflammatory exudation and poor granulation tissue growth.Additionally,there were increases in the expression levels of GRP78,CHOP and cas pase-12 proteins(P<0.01),as well as a significant elevation in the content of inflammatory factor IL-1β(P<0.01

关 键 词：温阳生肌膏 内质网应激 GRP78/CHOP通路 糖尿病难愈合创面 

分 类 号：R363.2[医药卫生—病理学] R587.1[医药卫生—基础医学] R289.6