摘要
目的:制备糖尿病大鼠背部全层皮肤缺损模型,观察复方ANBP对大鼠糖尿病创面愈合的影响和作用,从线粒体功能探讨复方ANBP对大鼠糖尿病创面的促愈机制。方法:将90只雄性SD大鼠随机分为空白组、模型组、复方ANBP组、贝复新组和烟酰胺单核苷酸(nicotinamide mononucleotide,NMN)组5组,每组16只。于3、7和14 d观察各组创面愈合情况后取材,分析创面愈合率,用HE和Masson染色观察局部组织病理变化,多重免疫荧光和Western blot检测三羧酸(tricarboxylic acid,TCA)循环相关酶丙酮酸脱氢酶E1α1亚基(pyruvate dehydrogenase E1 subunit alpha 1,PDHA1)、柠檬酸合酶(citrate synthase,CS)、异柠檬酸脱氢酶(isocitrate dehydrogenase,IDH1)和α-酮戊二酸脱氢酶(oxoglutarate dehydrogenase,OGDH)蛋白的表达情况,透射电镜观察创面组织线粒体数量和形态。结果:在糖尿病创面愈合进程中,空白组、复方ANBP组的组织形态学较模型组显著改善,在3、7和14 d空白组、复方ANBP组、贝复新组、NMN组创面愈合率显著升高(P<0.01)。与模型组相比,其余各组创面肉芽组织新生填补创面缺损,生成丰富的胶原纤维。在干预后3、7和14 d,与模型组对比,空白组、复方ANBP组、贝复新组、NMN组TCA循环相关酶PDHA1、CS、IDH1和OGDH荧光强度显著增强,表达数量增多。3、7和14 d模型组TCA循环相关酶水平显著降低(P<0.01),复方ANBP组、贝复新组、NMN组水平均显著升高(P<0.01)。复方ANBP干预后,复方ANBP组线粒体数量增多密度增加、线粒体形态改善空泡化率降低(P<0.05)。结论:复方ANBP可能是通过增加线粒体数量、改善线粒体形态,上调PDHA1、CS、IDH1和OGDH蛋白表达水平,改善线粒体功能,加速创面肉芽组织新生,从而达到促进大鼠糖尿病创面愈合的目的。
AIM:To investigate the effects of the compound ANBP on wound healing in diabetic rats and explore its mechanism of action.METHODS:Ninety male SD rats were randomly divided into blank,model,compound ANBP,Beifuxin,and nicotinamide mononucleotide(NMN)groups,with 16 rats in each group.Wound healing in each group was observed and samples were taken on days 3,7 and 14 to analyze the wound healing rate.Local histopathological changes were observed using HE and Masson staining.The expressions of pyruvate dehydrogenase E1 subunit alpha 1(PDHA1),citrate synthase(CS),isocitrate dehydrogenase(IDH1)and oxoglutarate dehydrogenase(OGDH)were detected through immunofluorescence and Western blot.The number and morphology of mitochondria in the wound tissue were observed using transmission electron microscopy.RESULTS:Histomorphological changes revealed significant improvement in diabetic wound healing in the blank and compound ANBP groups compared to that of the model group.The wound healing rates of the blank,compound ANBP,Beifuxin,and NMN groups were significantly increased on days 3,7,and 14(P<0.01).Compared to the model group,granulation tissue generation was higher in the other groups,covering the wound defect and producing abundant collagen fibers.At 3,7,and 14 days after intervention,the blank,compound ANBP,Beifuxin,and NMN groups showed significantly enhanced fluorescence intensities of TCA cycling-related enzymes PDHA1,CS,IDH1,and OGDH indicating increased expression of these enzymes.The levels of the TCA cycling-related enzymes were significantly increased(P<0.01)in the compound ANBP,Beifuxin and NMN groups but were significantly decreased(P<0.01)in the model group.An increase in the number and density of mitochondria and a decrease in the cavitation rate of mitochondria with improved morphology(P<0.05)was observed in the group treated with compound ANBP.CONCLUSION:Compound ANBP may increase the number of mitochondria,improve mitochondrial morphology and function,upregulate the expression levels of PDHA1,CS,IDH1,and OGDH proteins,and accelerate the regeneration of wound granulation tissue,thus promoting the healing of diabetic wounds in rats.
作者
王巍
侯倩
黄新灵
王君
邹梅林
丁雅容
郭乐天
谢晨磊
黄新宇
陈丽
周忠志
WANG Wei;HOU Qian;HUANG Xinling;WANG Jun;ZOU Meilin;DING Yarong;GUO Letian;XIE Chenlei;HUANG Xinyu;Chen Li;ZHOU Zhongzhi(College of Integrative Medicine,Hunan University of Chinese Medicine,Changsha 410208,China;Department of Burns and Wounds,The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410007,China;Key Labo-ratory of Vascular Biology,Hunan University of Traditional Chinese Medicine,Changsha 410208,China;Chinese Peo-ple's Liberation Army General Hospital,Beijing 100853,China;School of Medicine,Hunan University of Traditional Chi-nese Medicine,Changsha 410208,China;Hainan Provincial People's Hospital,Haikou 570311,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2024年第7期1283-1291,共9页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81973862,No.82074447)
湖南省卫生健康委重点项目(No.C202304039570)
海南省重点研发项目(No.ZDYF2023SHFZ123)
湖南中医药大学研究生创新项目(No.2022CX68)。
关键词
糖尿病
创面愈合
三羧酸循环
线粒体
diabetes mellitus
wound healing
tricarboxylic acid cycle
mitochondria
