摘要
目的：探讨抵当汤及其蕴含的泻热化瘀通络法对因核转录因子-кB（NF-κB）通路激活而导致的糖尿病（diabetes mellitus,DM）大鼠心肌炎症反应的影响。方法：选取130只体重达到220-270 g清洁级SD健康雄性大鼠,分为正常组（10只）和造模组（120只）,后者按55 mg·kg-1剂量一次性腹腔注射链脲佐菌素（streptozotocin,STZ）溶液建立DM模型。72 h后将造模成功的大鼠随机分为模型组,抵当汤高、中、低剂量组（1.08,0.72,0.54 g·kg^-1·d^-1）,吡咯啉烷二甲基硫脲组（pyrrolidine dithiocarbamate,PDTC组,0.15 g·kg^-1·d^-1）以及抵当-吡咯啉烷二甲基硫脲组（DP组1.08 g·kg^-1·d^-1抵当汤＋0.15 g·kg^-1·d^-1PDTC）,均采用相应药物1次/d进行灌胃干预。8周后,麻醉状态下腹主动脉取血处死动物,迅速摘取大鼠心脏,置于液氮中储存备用。以蛋白免疫印迹法（Western blot）检测心肌组织匀浆中转录NF-κB p65蛋白的表达,酶联免疫吸附法（ELISA）测定糖尿病心肌病（DCM）大鼠心肌组织匀浆肿瘤坏死因子-a（tumor necrosis factor-alpha,TNF-α）表达,免疫荧光检测心肌组织细胞黏附分子-1（intercellular cell adhesion molecule-1,ICAM-1）,巨噬细胞趋化性蛋白-1（macrophage chemotaxis protein-1,MCP-1）的表达。结果：与正常组比较,模型组NF-κB p65蛋白和TNF-α表达均明显升高;且模型组胞浆内观察到被染成绿色荧光的ICAM-1,MCP-1蛋白。与模型组比较,各治疗组NF-κB p65蛋白和TNF-α表达则明显降低;组间比较发现,DP组NF-κB p65蛋白表达与正常组最接近,抵当汤高剂量组TNF-α表达则较模型组降低更加显著;各治疗组胞浆内观察到被染成绿色荧光的ICAM-1,MCP-1蛋白不同程度减少,其中DP组被染成绿色的范围最少,与正常组最为接近。结论：泻热化瘀通络之抵当汤可通过抑制NF-κB通路的激活来减轻DM模型大鼠的心肌炎症反应。
Objective： To investigate the effect of Didangtang in purging heat,dispersing blood stasis and dredging collaterals on myocardial inflammation reaction of diabetes mellitus（ DM） rats by activating nuclear factor-κB（ NF-κB） pathway. Method： A total of 130 clean grade healthy male SD rats weighing between 220 g and270 g were selected and divided into the normal group（ 10 rats） and the model group（ 120 rats）. The rats of the model group were injected with 55 mg·kg-1 streptozotocin（ STZ） solution to establish the DM model. After 72 hours,the rats were randomly divided into the high-dose group,the model group,the low-dose group（ 1. 08,0. 72,0. 54 g·kg^-1·d^-1）,the pyrrolylene dimethyl thiourea group（ PDTC group 0. 15 g·kg^-1·d^-1） and the didangpyrrolidine dimethyl thiourea group（ DP group 1. 08 g·kg^-1·d^-1＋ 0. 15 g·kg^-1·d^-1 PDTC）,and intervened with the corresponding drugs once a day by gavage. After 8 weeks,the animals were put to death by drawing blood from the abdominal aorta in anesthesia. And then the rats＇ hearts were quickly removed and stored in liquid nitrogen.The NF-кB p65 protein expression in myocardium homogenate was detected by Western blot,the expression of tumor necrosis factor-α（ TNF-α） was detected by enzyme-linked immunosorbent assay（ ELISA） method,and the expressions of intercellular cell adhesion molecule-1（ ICAM-1） and macrophage chemotaxis protein-1（ MCP-1） in myocardium were detected by immunofluorescence. Result： The expressions of NF-кB p65 and TNF-α of the model group were significantly higher than those in the normal group. And we observed that ICAM-1 and MCP-1 proteins were dyed as green fluorescent in cytoplasm in the model group. Compared with the model group,the expressions of NF-кB p65 protein and TNF-α in the treatment groups were significantly lower. According to the inter-group comparison,the expression of NF-кB p65 protein in the DP group was the closest to that in the normal group,and the expression o
作者
储全根
王玲
张凯
潘小凤
CHU Quan-gen;WANG Ling;ZHANG Kai;PAN Xiao-feng(Anhui University of Chinese Medicine, Hefei 230038, Chin)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2018年第1期109-113,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
安徽省自然科学基金项目(1408085MKL32)
安徽中医药大学2016年新安医学教育部重点实验室建设项目(2016HACZ3042)
