基于SMAD4/hepcidin通路探讨脑泰方对大鼠局灶性脑缺血后皮质铁代谢的神经元保护作用  

Neuroprotective effect of Naotai Formula on cortical iron metabolism in rats after focal cerebral ischemia based on SMAD4/hepcidin pathway

作　　者：余清平 廖君 梅志刚 陈懿 王国佐[2] 王建君 葛金文 YU Qing-ping;LIAO Jun;MEI Zhi-gang;CHEN Yi;WANG Guo-zuo;WANG Jian-jun;GE Jin-wen(Medical College of Hunan University of Chinese Medicine,Changsha 410208,China;School of Integrated Traditional Chinese and Western Medicine,Hunan University of Chinese Medicine,Changsha 410208,China;Shaoyang University,Shaoyang 422000,China)

机构地区：[1]湖南中医药大学医学院,长沙410208 [2]湖南中医药大学中西医结合学院,长沙410208 [3]邵阳学院,邵阳422000

出　　处：《中华中医药杂志》2022年第8期4656-4660,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81774033,No.81774174);湖南省自然科学基金面上项目(No.2020JJ4467);湖南省教育厅科学研究重点项目(No.19A378)。

摘　　要：目的:研究脑泰方通过SMAD4/hepcidin通路干预局灶性脑缺血后皮质神经元铁代谢的相关机制。方法:将72只SD大鼠随机分为正常组,模型组,脑泰方低、中、高剂量组(3、9、27 g/kg),去铁酮组(125 mg/kg),每组12只。各组大鼠预处理灌胃给药连续3 d,大脑中动脉栓塞(MCAO)模型制备术后灌胃给药1 d。术后1 d取材,HE染色观察皮质神经元形态,ELISA检测血浆缺氧诱导因子α(HIF-1α)、铁调素(hepcidin)表达,QPCR检测SMAD4、hepcidin、Fpn mRNA表达,Western blot检测SMAD4、hepcidin、Fpn蛋白表达。结果:脑泰方中、高剂量组可见少量神经元空泡状及核染色质固缩,正常形态神经元较模型组明显增加。与正常组比较,模型组大鼠血浆内HIF-1α及hepcidin显著上调(P<0.05),大脑皮质SMAD4 mRNA及蛋白表达显著增加(P<0.01),hepcidin蛋白表达显著增加(P<0.01),Fpn蛋白表达显著降低(P<0.01);与模型组比较,各治疗组血浆HIF-1α及hepcidin显著下调(P<0.05),脑泰方中剂量组及去铁酮组SMAD4 mRNA表达显著降低(P<0.01),脑泰方低、高剂量组及去铁酮组Fpn mRNA表达显著增加(P<0.01);各治疗组SMAD4蛋白表达显著降低(P<0.05,P<0.01),脑泰方中、高剂量组及去铁酮组hepcidin蛋白表达显著减少(P<0.05,P<0.01),同时Fpn蛋白表达显著增加(P<0.05,P<0.01)。结论:脑泰方可能通过降低SMAD4及hepcidin的表达,增加Fpn的表达,促进神经元铁外排,减少铁聚集及氧化损伤,对脑缺血导致的铁超载损伤有一定保护作用。Objective:Research on the mechanism of Naotai Formula regulating iron metabolism via SMAD4/hepcidin pathway in cortical neurons after focal cerebral ischemia.Methods:A total of 72 SD rats were randomly divided into control group,model group,Naotai Formula low,medium and high dose groups(3,9,27 g/kg) and deferiprone(DFP) group(125 mg/kg),with 12 rats in each group.Each group was pretreated with gavage and administered for 3 consecutive days and middle cerebral artery occlusion(MCAO) model preparation for 1 day postoperative gavage administration.The samples were collected 1 day after operation.HE staining was used to detect the morphology of cortical neurons.The expression of hypoxia-inducible factor 1α(HIF-1α) and hepcidin in serum were detected by ELISA.Gene expression and protein of SMAD4,hepcidin and Fpn were detected by QPCR and Western blot.Protein of SMAD4,hepcidin and Fpn were detected by Western blot.Results:There were few vacuoles and nuclear pyknosis of neurons in Naotai Formula middle and high dose groups,and the number of normal neurons increased significantly compared with the model group.Compared with normal group,the expression of HIF-1α and hepcidin in plasma increased(P<0.05),SMAD4 mRNA and protein expression were significantly increased(P<0.01),hepcidin protein expression was significantly increased(P<0.01),and the expression of Fpn protein was significantly decreased(P<0.01) in model group.Compared with the model group,the expression of HIF-1α and hepcidin in plasma decreased significantly in all treatment groups(P<0.05),the expression of SMAD4 mRNA was significantly decreased in Naotai Formula and medium dose group and DFP group(P<0.01),and the expression of Fpn mRNA was significantly increased in Naotai Formula low and high dose group and DFP group(P<0.01);The protein expression of SMAD4 was significantly decreased in each treatment group(P<0.05,P<0.01),the expression of hepcidin was significantly decreased in Naotai Formula medium and high dose group and DFP group(P<0.05,P<0.01),and the

关 键 词：脑泰方 脑缺血 铁代谢 SMAD4 铁调素 膜铁转运蛋白 

分 类 号：R285.5[医药卫生—中药学]