黄芪-当归通过TGF-β_(1)/Smad2信号途径抑制兔血管内膜增生模型外膜成纤维细胞的活化作用  

Astragali Radix Combined with Angelicae Sinensis Radix Inhibits Activation of Adventitial Fibroblasts in Rabbit Model of Intimal Hyperplasia via TGF-β_(1)/Smad2 Signaling Pathway

作　　者：李霞[1,2] 曹旺 邓常清 LI Xia;CAO Wang;DENG Changqing(Key Laboratory of Hunan Province for Prevention and Treatment of Integrated Traditional Chinese and Western Medicine on Cardiocerebral Diseases,Hunan University of Chinese Medicine,Changsha 410028,China;Hunan Academy of Chinese Medicine,Changsha 410028,China)

机构地区：[1]湖南中医药大学中西医结合心脑疾病防治湖南省重点实验室,长沙410028 [2]湖南省中医药研究院,长沙410028

出　　处：《中国实验方剂学杂志》2023年第3期61-69,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81874406)。

摘　　要：目的:探讨黄芪-当归配伍对兔血管内膜增生模型血管外膜成纤维细胞(VAF)活化的影响及其作用机制。方法:清洁级家兔,雌雄各半,随机分为假手术组、模型组、单用黄芪组(1 g·kg^(-1))、单用当归组(1 g·kg^(-1))、黄芪-当归1∶1组(黄芪1 g·kg^(-1)+当归1 g·kg^(-1))、黄芪-当归1∶5组(黄芪1 g·kg^(-1)+当归5 g·kg^(-1))、黄芪-当归5∶1组(黄芪5 g·kg^(-1)+当归1 g·kg^(-1))、阿托伐他汀钙组(5 mg·kg^(-1)),采用颈总动脉套管复合高胆固醇饮食制备家兔血管内膜增生模型,假手术组只进行分离操作,不进行套管。术后第1天开始假手术组喂饲普通饲料,其余各组喂饲高胆固醇饲料(2%胆固醇+3%橄榄油+95%普通饲料),同时各用药组给予相应药物灌胃,假手术组、模型组给予等体积蒸馏水灌胃,持续给药28 d。实验结束后腹主动脉采血,马松(Masson)染色观察血管外膜形态学变化,免疫组化法检测外膜局部促炎因子肿瘤坏死因子-α(TNF-α)及白细胞介素-1β(IL-1β)的表达,免疫荧光双标法测定外膜成纤维细胞标记物波形蛋白(Vimentin)及肌成纤维细胞标记物α-平滑肌肌动蛋白(α-SMA)表达反映VAF活化;蛋白免疫印迹法(Western blot)检测相关通路蛋白转化生长因子-β_(1)(TGF-β_(1))、Smad2、磷酸化(p)-Smad2表达。结果:与假手术组比较,模型组血管外膜增生明显,损伤血管局部TNF-α、IL-1β表达显著升高,受损段血管α-SMA、Vimentin、TGF-β_(1)、Smad2、p-Smad2表达均明显升高(P<0.05,P<0.01);与模型组比较,黄芪-当归1∶1、5∶1组可使血管外膜增生减轻,血管局部TNF-α、IL-1β表达降低,α-SMA、Vimentin表达下调,并且可降低TGF-β_(1)、Smad2、p-Smad2表达(P<0.05,P<0.01)。结论:黄芪当归配伍可抑制血管内膜增生模型的外膜增生,抑制外膜成纤维细胞向肌成纤维细胞转化,减轻血管局部炎症反应,以黄芪-当归1∶1、5∶1组的作用明显。其作用机制可能与黄芪�Objective:To observe the effect of Astragali Radix combined with Angelicae Sinensis Radix on the activation of adventitial fibroblasts in the rabbit model of intimal hyperplasia and decipher the underlying mechanism.Method:Clean white rabbits of the same number of males and females were selected.They were randomly assigned into a sham operation group,a model group,an Astragali Radix group(1 g·kg^(-1)),an Angelicae Sinensis Radix group(1 g·kg^(-1)),and Astragali Radix-Angelicae Sinensis Radix groups at the ratios of 1∶1(Astragali Radix 1 g·kg^(-1)+Angelicae Sinensis Radix 1 g·kg^(-1)),1∶5(Astragali Radix1 g·kg^(-1)+Angelicae Sinensis Radix 5 g·kg^(-1)),and 5∶1(Astragali Radix 5 g·kg^(-1)+Angelicae Sinensis Radix1 g·kg^(-1)),and an atorvastatin calcium group(5 mg·kg^(-1)).The rabbit model of intimal hyperplasia was established by common carotid artery cannula combined with high-cholesterol diet.The sham operation group only underwent the separation operation and not the cannula.The sham operation group had been fed with normal diet and the other groups with high-cholesterol diet(2%cholesterol+3%olive oil+95%common diet)since the first day after operation.At the same time,drugs were administrated in corresponding groups,and the sham operation group and model group were administrated with distilled water(15 mL·kg^(-1))for 28 days.At the end of the experiment,blood was collected from abdominal aorta,and the morphological changes of adventitia were observed by Masson staining.The expression of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)in adventitia was detected by immunohistochemical method.The expression of adventitial fibroblast marker vimentin and myofibroblast markerα-smooth muscle actin(α-SMA),which can reflect the activation of adventitial fibroblasts,was detected by immunofluorescence double staining.Western blot was employed to determine the protein levels of transforming growth factor-β_(1)(TGF-β_(1)),Smad2,and p-Smad2.Result:Compared with the normal group,the modeling caus

关 键 词：黄芪 当归 血管外膜成纤维细胞 肌成纤维细胞 转化生长因子-β_(1)(TGF-β_(1))/Smad2信号通路 

分 类 号：R2-0[医药卫生—中医学] R33R289R543.12