MEBT/MEBO调控肌成纤维细胞促进糖尿病难愈合创面修复的机制研究  

Mechanism study of MEBT/MEBO regulating myofibroblasts to promote healing of diabetic refractory wounds

作　　者：贺佐分 田馨如 龚元勋 黄金梅 杨雅量 葛星月 岑丽君 郭文闻 唐乾利[3,4] He Zuofen;Tian Xinru;Gong Yuanxun;Huang Jinmei;Yang Yaliang;Ge Xingyue;Cen Lijun;Guo Wenwen;Tang Qianli(Graduate School,Youjiang Medical University for Nationalities,Baise 533000,Guangxi,China;Graduate School,Guangxi University of Chinese Medicine,Nanning 530023,Guangxi,China;The Affiliated Hospital of Youjiang Medical University for Nationalities,Baise 533000,Guangxi,China;Guangxi University Key Laboratory of High Incidence Prevention and Control Research in Western Guangxi,Baise 533000,Guangxi,China)

机构地区：[1]右江民族医学院研究生学院,广西百色533000 [2]广西中医药大学研究生院,广西南宁530023 [3]右江民族医学院附属医院,广西百色533000 [4]广西高校桂西地区高发病防治研究重点实验室,广西百色533000

出　　处：《右江民族医学院学报》2023年第2期182-189,共8页Journal of Youjiang Medical University for Nationalities

基　　金：国家自然科学基金面上项目(81774327);广西医学高层次骨干人才培养“139”计划项目资助(桂卫科教发〔2018〕22号);右江民族医学院研究生教育创新计划(桂学位〔2021〕5号)(YCSW20211334)。

摘　　要：目的探讨湿润暴露疗法/湿润烧伤膏(moist exposed burn therapy/moist exposed burn ointment,MEBT/MEBO)是否通过调控糖尿病难愈合创面肌成纤维细胞,影响α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)和Ⅰ型胶原蛋白(collagenⅠ,COL-1)的表达水平,从而促进创面愈合。方法90只SPF级成年雄性Wistar大鼠以随机数表法分为5组,空白组、对照组、模型组、MEBO组、贝复新组,每组18只,适应性喂养7 d后构建糖尿病模型,成模后统一制备创面。创面形成后第0天、第3天、第7天、第14天拍照记录创面愈合情况,每组随机取6只大鼠创面组织,进行HE、Masson染色以观察各组创面组织病理形态变化,免疫荧光观察创面组织α-SMA、COL-1的分布及表达情况,Western Blot检测创面组织α-SMA、COL-1的蛋白表达水平。结果①同一时间点组织学观察各组病理形态改善均优于模型组,治疗后期创面愈合率MEBO组、贝复新组优于模型组。②WB:与对照组比较,创面发生第3天、第7天、第14天过程中,模型组创面组织α-SMA、COL-1蛋白表达均明显下降(P<0.05);与模型组比较,创面第7天、第14天,MEBO组、贝复新组创面组织α-SMA、COL-1表达显著升高(P<0.05)。③免疫荧光:与对照组比较,第3天、第7天、第14天的模型组α-SMA蛋白表达均较低,其中第7天时具有显著性差异(P<0.05);模型组COL-1蛋白表达均显著偏低(P<0.05)。与模型组比较,第7天、第14天MEBO组、贝复新组α-SMA表达均较高(P<0.05),两用药组COL-1表达也较高,而贝复新组与模型组的比较差异有统计学意义(P<0.05),MEBO组COL-1表达介于模型组与对照组之间,但与模型组的比较差异不明显(P>0.05)。结论MEBT/MEBO可能通过提高α-SMA和COL-1的蛋白表达水平,促进肌成纤维细胞转化及细胞外基质的沉积,从而加速糖尿病难愈合创面修复。Objective To explore whether moist exposed burn therapy/moist exposed burn ointment(MEBT/MEBO)regulates myofibroblasts in diabetic refractory wounds and affects the expression levels ofα-smooth muscle actin(α-SMA)and typeⅠcollagen(COL-1),thus promoting wound healing.Methods Ninety SPF-grade adult male Wistar rats were randomly divided into 5 groups:the blank group,the control group,the model group,MEBO group,and rb-bFGF group,with 18 rats in each group.After 7 days of adaptive feeding,diabetic models were established and wounds were created uniformly.The wound healing status was photographed and recorded on days 0,3,7 and 14 after wound formation.Six rats from each group were randomly selected for HE and Masson staining to observe the pathological morphological changes of wound tissues in each group.Immunofluorescence was used to observe the distribution and expression ofα-SMA and COL-1 in wound tissues.Western Blot was used to detect the protein expression levels ofα-SMA and COL-1 in wound tissues.Results①Histological observations at the same time point showed that the pathological morphological changes in all groups were better than those in the model group,and the wound healing rate in the later stage of treatment was higher in the MEBO group and rb-bFGF group than in the model group.②WB:compared with the control group,the protein expression ofα-SMA and COL-1 in wound tissues of the model group significantly decreased on day 3,7,and 14 after wound formation(P<0.05).Compared with the model group,the expression ofα-SMA and COL-1 in wound tissues of the MEBO group and rb-bFGF group significantly increased on day 7 and 14(P<0.05).③Immunofluorescence:compared with the control group,the protein expression ofα-SMA in the model group was lower on day 3,7,and 14,with significant differences on day 7(P<0.05).The expression of COL-1 in the model group was significantly lower(P<0.05).Compared with the model group,the expression ofα-SMA in the MEBO group and rb-bFGF groups was higher on day 7 and 14(P<0.05)

关 键 词：湿润暴露疗法/湿润烧伤膏 糖尿病难愈合创面 α-平滑肌肌动蛋白 Ⅰ型胶原蛋白 肌成纤维细胞 

分 类 号：R587.1[医药卫生—内分泌]