拔毒生肌散调控Keap1/Nrf2通路促进糖尿病大鼠皮肤溃疡愈合的机制研究  

Study on Mechanism of Badu Shengji Powder on Regulating Keap1/Nrf2 Pathway to Promote Skin Ulcer Healing in diabetes Rats

作　　者：王双勋[1,2] 李大勇[2] WANG Shuangxun;LI Dayong(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,China)

机构地区：[1]辽宁中医药大学,沈阳110847 [2]辽宁中医药大学附属医院,沈阳110032

出　　处：《中国中医基础医学杂志》2023年第10期1645-1649,共5页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：中国人口福利基金项目(FY202107140413);国家重点研发计划(2019YFC1709303)。

摘　　要：目的 观察拔毒生肌散对糖尿病大鼠皮肤溃疡肉芽组织Kelch样环氧氯丙烷相关蛋白(Kelch-like ECH-associated protein,Keap)-1/核因子E2相关因子(nuclear factor-E2-related factor 2,Nrf2)通路的影响,探讨拔毒生肌散促进糖尿病大鼠皮肤溃疡愈合的作用机制。方法 将60只大鼠随机分为正常对照组、模型组、拔毒生肌散组、一效膏组、清创胶组,每组12只。除正常组外,其他4组大鼠采用腹腔注射链脲佐菌素(streptozocin,STZ)结合喂食高脂高糖饲料的方法建立糖尿病模型,在此基础上建立背部皮肤溃疡模型。外用相应药物治疗14 d后,评估并比较各组大鼠溃疡面面积及愈合率;苏木素-伊红(hematoxylin and eosin,HE)染色、Masson染色、CD34免疫组化检测大鼠肉芽组织病理变化;Western blot、荧光定量PCR分别检测大鼠肉芽组织Keap1、Nrf2蛋白和m RNA表达水平。结果 与正常对照组比较,模型组大鼠溃疡面愈合率、胶原纤维阳性面积、血管内皮细胞阳性率、Nrf2蛋白和m RNA表达显著降低(P<0.01),Keap1蛋白和m RNA表达显著升高(P <0.01),炎性细胞浸润较多,胶原纤维蓝染少,排列紊乱,分布稀疏,CD34标记新生血管较少,管径较细。与模型组比较,拔毒生肌散组大鼠溃疡面愈合率、胶原纤维阳性面积、血管内皮细胞阳性率、Nrf2蛋白和m RNA表达显著升高(P <0.01),Keap1蛋白和m RNA表达显著降低(P <0.01),炎性细胞浸润较少,胶原纤维蓝染增多,排列相对整齐有序,CD34标记新生血管较多,管径较粗。各治疗组之间溃疡面愈合率比较差异无统计学意义(P>0.05)。结论 拔毒生肌散能够通过抑制溃疡面炎症、促进胶原纤维沉积和血管新生以及调控Keap1/Nrf2通路促进糖尿病大鼠皮肤溃疡的愈合。Objective To observe the effect of Badu Shengji powder on Kelch-like ECH-associated protein(Keap)-1/nuclear factor-E2-related factor 2(Nrf2)pathway in granulation tissue of skin ulcer in diabetes rats,and to explore the mechanism of Badu Shengji powder in promoting the healing of skin ulcer in diabetes rats.Methods 60 rats were randomly divided into normal control group,model group,Badu Shengji powder group,Yixiao ointment group and Qingchuang glue group,with 12 rats in each group.In addition to the normal group,intraperitoneal injection of STZ combined with feeding high fat and high sugar diet method was used to establish the diabetes rat model.On the basis of diabetes rat model,the diabetes skin ulcer model was further established.After 14 days of external application of the corresponding drugs,the ulcer area of rats in each group was measured and the healing rate was calculated.HE staining,Masson staining and CD34 immunohistochemistry were used to detect the pathological changes of rat granulation tissue.Western blot method and fluorescent quantitative PCR were used to detect the protein level and mRNA expression of Keap1 and Nrf2 in rat granulation tissue.Results Compared with the normal group,the healing rate of ulcer surface,the positive area of collagen fibers,the positive rate of vascular endothelial cells,the protein and mRNA expression of Nrf2 were significantly decreased,while the protein and mRNA expression of Keap1 were significantly increased in the rats of model group(P<0.01).There were more inflammatory cell infiltration,collagen fibers with less blue staining,disordered arrangement,and sparse distribution in the rats of model group.CD34 marked neovascularization was less and the diameter was smaller in the rats of model group.Compared with the model group,the healing rate of ulcer surface,the positive area of collagen fibers,the positive rate of vascular endothelial cells,the protein amount of Nrf2 and mRNA expression were significantly increased(P<0.01),while the protein and mRNA expression of K

关 键 词：糖尿病 皮肤溃疡 Keap1/Nrf2通路 拔毒生肌散 

分 类 号：R275.9[医药卫生—中西医结合]