摘要
目的观察黄芪提取液对糖尿病溃疡的治疗效果,探讨其促进糖尿病溃疡创面愈合的作用机制。方法雄性Wistar大鼠随机分为正常对照组、模型组、黄芪提取液低、中、高剂量组(1.35、2.7、5.4 g·kg-1),采用大剂量链脲佐菌素腹腔注射和外科方法制备糖尿病溃疡模型。于溃疡造模后第2天各组分别经口灌胃给药及常规换药,观察给药第3天、7天、14天、21天各组大鼠创面愈合情况;分别于给药后第7、14、21天,应用Real Time-PCR检测大鼠创面组织Wnt1、Wnt3a、β-Catenin、GSK-3βmRNA的表达,采用Western Blot检测β-Catenin、GSK-3β、C-myc、CyclinD1及β-Catenin核内表达水平,以免疫组化法观察创面组织β1整合素、K19、PCNA的表达情况。结果与对照组比较,模型组、各用药组创面造模后第3天、7天、14天创面愈合率均明显降低(P<0.01),模型组、低剂量组创面愈合时间明显延长(P<0.01);与模型组比较,高剂量组给药后第14、21天创面愈合率显著提高(P<0.01),创面愈合时间缩短(P<0.05);与模型组比较,高剂量组给药后第7天、14天、21天创面组织Wnt1、Wnt3a、β-catenin mRNA表达加强(P<0.05),β-Catenin、C-myc及CyclinD1蛋白表达增加(P<0.05),GSK3βmRNA及蛋白表达均降低(P<0.05),β1整合素、K19、PCNA的表达增加(P<0.05)。结论黄芪提取液能促进糖尿病溃疡愈合,其机制可能与通过对Wnt/β-catenin信号通路的调节,促进表皮干细胞的增殖分化,加快创面上皮化的进程有关。
Objective To observe the therapeutic effect of Astragalus extract on diabetic ulcer and explore its mechanism of promoting wound healing.Methods Male Wistar rats were randomly divided into normal control group,model group,astragalus extract low,medium and high dose group(1.35,2.7,5.4 g·kg-1).Diabetic ulcer model was made by intraperitoneal injection of high-dose streptozotocin and surgical method.On the second day after the ulcer model was made,each group was given medicine by gavage and routine surgical dressing change,the wound healing of rats in each group was observed on the 3 rd,7 th,14 th and 21 st day after administration;real time PCR was used to detect to expression of Wnt1,Wnt3 a,β-Catenin and GSK-3βmRNA on the 7 th,14 th and 21 st day after administration.Western blot was used to detect the expression ofβ-Catenin,expression ofβ-Catenin in nucleus,GSK-3β,C-myc and CyclinD1.Immunohistochemistry was used to observe the expression ofβ1 integrin,K19,PCNA in wound tissue.Results Compared with the control group,the wound healing rate of the model group and each medication group decreased on the 3 rd,7 th and 14 th day(P<0.01),and the wound healing time of the model group and the low dose group prolonged significantly(P<0.01).Compared with the model group,the wound healing rate of the high dose group increased significantly on the 14 th and 21 st day(P<0.01),and the wound healing time shortened(P<0.05).Compared with the model group,expression of Wnt1,Wnt3 a andβ-Catenin mRNA was increased on the 7 th,14 th and 21 st day in high dose group(P<0.05),expression ofβ-Catenin,C-myc and CyclinD1 protein was increased(P<0.05),expression of GSK3βmRNA and GSK3βprotein was decreased(P<0.05),expression ofβ1 integrin,K19 and PCNA was increased(P<0.05).Conclusion Astragalus extract can promote the healing of diabetic ulcer.Its mechanism may be related to the regulation of Wnt/β-Catenin signal pathway,the promotion of proliferation and differentiation of epidermal stem cells,and the acceleration of the process of
作者
仇莲胤
阙华发
屈可伸
孙伟
郭树豫
李阳
Qiu Lianyin;Que Huafa;Qu Keshen;Sun Wei;Guo Shuyu;Li Yang(Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;LongHua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2020年第7期2522-2530,共9页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
国家自然科学基金面上项目(81774317):补虚化瘀中药调控microRNA促进糖尿病性溃疡创面愈合的机制研究,负责人:阙华发
上海中医药大学高峰高原创新团队(A1-N1920501010212),负责人:阙华发
