局部应用重组人EGF对糖尿病大鼠烫伤创面EGF受体及其mRNA表达的影响  

EFFECT OF TOPICAL EXTERNAL ADMINISTRATION OF RECOMBINANT HUMAN EPIDERMAL GROWTH FACTOR ON EXPESSION OF EPIDERMAL GROWTH FACTOR RECEPTOR AND ITS mRNA IN SCALD WOUND OF DIABETES MELLITUS RAT

作　　者：宗守凯[1] 梁自乾[1] 欧邦军[1] 

机构地区：[1]广西医科大学第一附属医院烧伤整形外科,南宁530021

出　　处：《中国修复重建外科杂志》2010年第2期150-155,共6页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：广西壮族自治区科技厅资助项目(0542093);广西研究生教育创新计划基金(2007105981002M32)~~

摘　　要：目的EGF对创面修复具有积极作用,EGF受体(EGF receptor,EGFR)可调节皮肤创面愈合,通过建立糖尿病大鼠烫伤创面模型,观察局部应用重组人EGF(recombinant human EGF,rhEGF)联合血糖控制对创面EGFR及其mRNA的影响,探讨rhEGF促进创面愈合的机制。方法136只清洁级雄性Wistar大鼠,体重(188.57±6.59)g,随机分为A、B、C、D组(n=34)。A、B、C组大鼠腹腔内一次性注射链脲佐菌素(60mg/kg)制备糖尿病模型,D组于对应时间点注射等量柠檬酸缓冲液,作为正常对照组。糖尿病模型成功制备后8周,取4组大鼠背部去毛后置于80℃热水6s,制备背部深Ⅱ度烫伤模型。A组于烫伤模型制备前1周开始控制血糖至D组水平;模型制备24h内创面清创后喷洒150U/cm2rhEGF,含1%磺胺嘧啶银霜剂辅料缝扎固定;B组不控制血糖,伤后处理同A组;C组同A组方法控制血糖,模型制备后24h内采用含1%磺胺嘧啶银霜剂辅料缝扎固定;D组伤后处理同A组。烫伤后3、7、11、15、21d测定各组大鼠创面愈合率;并于烫伤后1、3、5、7、11、15、21d,取创面皮肤组织采用mRNA原位杂交法检测EGFR mRNA表达、免疫组织化学染色及Western blot检测EGFR蛋白表达情况结果各组大鼠均存活至实验完成。烫伤后3d各组创面愈合率比较,差异无统计学意义(P>0.05);7、11、15、21d,A、D组高于B、C组(P<0.05);A、D组间及B、C组间比较,差异均无统计学意义(P>0.05)。mRNA原位杂交观察显示,烫伤后各组均有EGFR mRNA表达,主要分布于真皮的成纤维细胞、毛细血管内皮细胞及创缘残存的皮肤附属器上皮中;A、B、C、D组分别于烫伤后5、7、7、11d表达达峰值,且A、D组峰值较B、C组高,差异有统计学意义(P<0.05);A、D组间及B、C组间差异无统计学意义(P>0.05)。免疫组织化学染色及Western blot检测均可见各组EGFR蛋白表达,并逐渐增强,A、B、C、D组分别于烫伤后7、7、11、11d达峰值,A、D组间及B、C组间比较差异�Objective To investigate the effect of topical external administration of recombinant human epidermal growth factor（rhEGF）when controlling blood sugar on expression of epidermal growth factor receptor（EGFR）and EGFR mRNA of wound in diabetes mellitus（DM）combined with scald.Methods A total of 136 male Wistar rats weighing（188.57±6.59）g were randamly divided into 4 groups（groups A,B,C,and D,n=34）.The rats was made DM model by intraperitoneal injected 60 mg/kg streptozocin in groups A,B,and C;rats were injected buffer alone in group D as control group.After 8 weeks,the rats of 4 groups were placed in 80℃hot water for 6 seconds for preparation of the back deep II degree scald model.In group A,the blood sugar level was controlled at the level of group D 1 week before scald model;within 24 hours after models preparation,rhEGF was sprayed on wound at 150 U/cm2.In group B,the rats were given the same treatment as group A except not controlling blood sugar.In group C,the blood sugar was controlled as group A and wound was suture fixation with 1%silver sulfadiazine cream at 24 hours after the model.In group D,the same treatment as group A was given after injury.The healing rate of the wound was detected at 3,7,11,15,and 21 days after injury;the EGFR mRNA expression was determined by mRNA hybridization in situ,and the EGFR protein expression was deterimined by immunohistochemistry and Western blot at 1,3,5,7,11,15,and 21 days.Results All the rats survived at the end of experiment.There was no significant difference in the healing rate of the wound among the 4 groups at 3 days（P0.05）.The healing rate of the wound was significantly higher in groups A and D than in groups B and C（P0.05）at 7,11,15,and 21 days.The expession of EGFR mRNA in 4 groups was observed by hybridization in situ,which mainly distributed in the dermal fibroblasts,capillary endothelial cells and remnants of skin and wound edge epithelium of the subsidiary;the expessions reached the peak at 5 days in group A,at 7 days in groups

关 键 词：糖尿病 烫伤创面 创面愈合 重组人EGF EGF受体 大鼠 

分 类 号：R587.1[医药卫生—内分泌] R644[医药卫生—内科学]