解毒生肌膏调控GSK-3β/β-catenin减轻炎症反应促糖尿病SD大鼠慢性创面修复的影响  

Jiedu-Shengji ointment regulates GSK-3β/β-catenin to reduce inflammatory response to promote chronic wound healing in diabetic SD rats

作　　者：袁忠行 陈丽 丁雅容 王巍 王宇萌 郑时旭 洪恺祺 周忠志[3] Yuan Zhongheng;Chen Li;Ding Yarong;Wang Wei;Wang Yumeng;Zheng Shixu;Hong Kaiqi;Zhou Zhongzhi(School of Integrated Traditional Chinese and Western Medicine,Hunan University of Chinese Medicine;Medical College,Hunan University of Chinese Medicine;Department of Burn&Skin Ulcer Plastic Surgery,The First Affiliated Hospital of Hunan University of Chinese Medicine)

机构地区：[1]湖南中医药大学中西医结合学院,长沙410208 [2]湖南中医药大学医学院,长沙410208 [3]湖南中医药大学第一附属医院烧伤疮疡整形科,长沙410007

出　　处：《重庆医科大学学报》2023年第9期1069-1077,共9页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(编号:81973862);湖南省卫生健康委科研资助项目(编号:D202304039371,D202304039377);湖南省研究生科研创新资助项目(编号:CX20210722、CX20220811)。

摘　　要：目的:从GSK-3β/β-catenin角度探讨炎症反应对糖尿病SD大鼠慢性创面修复的影响。方法:使用高脂饲料喂养联合腹腔注射链脲霉素(Streptozocin,STZ)及手术制备全层皮肤缺损的方法建立糖尿病SD大鼠创面模型,成模后随机分为模型组(n=15),磺胺嘧啶银组(n=15)及解毒生肌膏组(n=15),并设立正常创面组(n=15)。磺胺嘧啶银组创面和解毒生肌膏组创面予以磺胺嘧啶银软膏和解毒生肌膏外敷,每日1次,模型组与正常创面组不予干预。给药后第3天、7天和14天肉眼观察各组大鼠的创面愈合情况,计算创面愈合率;HE观察组织基本形态;Masson检测胶原生成情况;超微电镜观察内质网结构差异;免疫组织化学法检测肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)和白细胞介素-6(Interleukin-6,IL-6)表达;蛋白质免疫印迹法检测糖原合成酶激酶-3β(glycogen synthase kinase-3β,GSK-3β)蛋白和β-连环蛋白(β-catenin)蛋白表达。结果:给药后3天解毒生肌膏组创面愈合率高于模型组(P<0.05);给药后14天解毒生肌膏组创面愈合率高于磺胺嘧啶银组(P<0.05)。HE结果:给药后3、7天解毒生肌膏组创面较其他组肿胀程度较小,毛细血管数量丰富;14天,解毒生肌膏组和正常创面组出现皮肤附属器新生,组织结构完整度好。Masson结果:解毒生肌膏组较其他各组胶原纤维分布均匀,排列结构较为整齐,新生胶原纤维更为粗壮,分布均匀,排列整齐。电镜结果:解毒生肌膏组内质网丰富,数量较多,形态结构趋于正常。免疫组织化学法结果:与模型组相比,各组TNF-α、IL-6表达水平明显降低;解毒生肌膏组TNF-α表达水平低于正常创面组;解毒生肌膏组TNF-α、IL-6表达低于磺胺嘧啶银组。Western blot法结果显示:与正常创面组相比,各组β-catenin表达量均上调(P<0.05),解毒生肌膏组上调表达量最少;与正常创面组相比,各组GSK-3β蛋白表达含量明显下调(P<0.05);给药后7�Objective:To investigate the effects of inflammatory response on the repair of chronic wounds in Sprague-Dawley(SD)rats with diabetes from the perspective of glycogen synthase kinase-3β(GSK-3β)/β-catenin.Methods:A wound model in diabetic SD rats was established by high-fat feeding combined with intraperitoneal injection of streptozocin and surgical preparation of full-thickness skin defects.The model rats were randomly divided into model group(n=15),silver sulfadiazine group(n=15),and Jiedu-Shengji oint⁃ment group(n=15),with normal wound group(n=15)set as a con⁃trol.The wounds in the silver sulfadiazine group and Jiedu-Shengji ointment group were externally treated with silver sulfadiazine oint⁃ment and Jiedu-Shengji ointment,respectively,once daily,with no intervention in the model group and normal wound group.On the 3rd,7th,and 14th days after ointment administration,the wound healing status of each group was examined to calculate the wound healing rate. The basic tissue morphology was examined with hematoxylin-eosin(HE) staining. Collagen production was detected byMasson. The structure of endoplasmic reticulum was observed using a transmission electron microscope. The expression of tumor necrosisfactor- α(TNF- α) and interleukin-6(IL-6) was measured by immunohistochemistry. The protein expression of GSK-3β and β-catenin was determined by Western blot. Results:On day 3,the wound healing rate of the Jiedu-Shengji ointment group was signifi⁃cantly higher than that of the model group(P<0.05). On day 14,the wound healing rate of the Jiedu-Shengji ointment group was signifi⁃cantly higher than that of the silver sulfadiazine group(P<0.05). HE staining revealed less swelling and more abundant capillaries inthe wounds of the Jiedu-Shengji ointment group as compared with the other groups on days 3 and 7 and new skin appendages andintact tissue structure in the wounds of the Jiedu-Shengji ointment group and normal wound group on day 14. Masson staining revealedevenly distributed and orderly organized col

关 键 词：糖尿病慢性创面 解毒生肌膏 糖原合成酶激酶-3Β Β-连环蛋白 炎症因子 

分 类 号：R2-031[医药卫生—中西医结合]