温肺降浊方通过调节Nrf2/Keap 1-GPx4信号通路抑制铁死亡改善血管性痴呆模型大鼠认知功能障碍的研究  

Wenfei Jiangzhuo(温肺降浊)Formula Inhibits Ferroptosis by Regulating Nrf2/Keap 1-GPx4 Signaling Pathway to Treat Cognitive Impairment in the Rat Model of Vascular Dementia

作　　者：张鼎 李方存[1] 姜明贺 周珂青 秦红玲[2] 陈炜[1] 胡跃强[2] ZHANG Ding;LI Fangcun;JIANG Minghe;ZHOU Keqing;QIN Hongling;CEHN Wei;HU Yueqiang(Guangxi University of Chinese Medicine,Nanning 530001;The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023)

机构地区：[1]广西中医药大学,南宁530001 [2]广西中医药大学第一附属医院,南宁530023

出　　处：《中药药理与临床》2024年第10期28-33,共6页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金项目(编号:82260904);广西中医脑病临床研究中心项目(编号:桂科AD20238028);国家中医药管理局高水平重点学科-中医内科学项目(编号:ZYYZDXK-2023166);广西中医药大学“歧黄工程”高层次人才团队项目(编号:202410);广西中医药大学博士研究生创新项目(编号:YCBXJ2023010);广西中医药重点学科建设项目(编号:GZXK-Z-20-13)。

摘　　要：目的:观察温肺降浊方对血管性痴呆模型大鼠的影响,探究其可能的作用机制。方法:制备永久性双侧颈总动脉闭塞(2VO)大鼠模型,使用Ferrostatin-1抑制剂和温肺降浊方2.95、5.9、11.8 g/kg干预,持续2个月。观察成模后大鼠学习认知能力,HE染色观察海马病理情况,普鲁士蓝染色、免疫组化、qRT-PCR和Western Blot法检测大鼠海马组织中铁沉积和GPx4、Nrf2、HO-1、Keap 1、Cox2 mRNA及蛋白的表达情况。结果:与假手术对照组比较,模型对照组大鼠逃避潜伏期延长、穿越平台次数明显缩短(P<0.05),海马细胞出现明显水肿及棕蓝色铁颗粒沉积;海马组织中GPx4、Nrf2、HO-1蛋白及mRNA的表达下调(P<0.05),Keap 1、Cox2蛋白及mRNA的表达上调(P<0.05);与模型对照组比较,Ferrostatin-1抑制剂和温肺降浊方各剂量组大鼠逃避潜伏期缩短、穿越平台次数明显增加(P<0.05),海马组织细胞水肿化减少;GPx4、Nrf2、HO-1蛋白及mRNA的表达上调(P<0.05),Keap 1、Cox2蛋白及mRNA的表达下调(P<0.05)。结论:温肺降浊方可能通过调节Nrf2/Keap 1/GPx4信号通路相关蛋白抑制铁死亡,从而减少大脑海马组织中铁颗粒的沉积,从而改善大脑认知情况,以温肺降浊方11.8 g/kg组作用最佳。Objective:To observe the effect of Wenfei Jiangzhuo(温肺降浊)Formula on vascular dementia in rats and decipher the possible mechanism.Methods:The rat model of 2-vessel occlusion was established and then treated with ferrostatin-1 inhibitor and Wenfei Jiangzhuo Formula at 2.95,5.9,and 11.8 g/kg for 2 months.The cognitive learning ability of the rats was observed after modeling.Hematoxylin-eosin staining and Prussian blue staining were employed to detect the iron deposition in the hippocampus.Immunohistochemistry,qRT-PCR,and Western blotting were employed to determine the expression of glutathione peroxidase 4(GPx4),nuclear factor-erythroid 2 related factor 2(Nrf2),heme oxygenase 1(HO-1),Kelch-like ECH-associated protein 1(Keap 1),and cyclooxygenase-2(Cox2)in the hippocampus.Results:Compared with the sham group,the model group showed prolonged escape latency and reduced platform crossings(P<0.05),obvious edema and deposition of brown-blue iron particles in the hippocampus cells,down-regulated expression of GPx4,Nrf2,and HO-1(P<0.05),and up-regulated expression of Keap 1 and Cox2(P<0.05)in the hippocampus.Compared with the model group,the ferrostatin-1 inhibitor and Wenfei Jiangzhuo Formula shortened the escape latency and increased the platform crossings(P<0.05),reduced the edema in the hippocampus,up-regulated the expression of GPx4,Nrf2,and HO-1(P<0.05),and down-regulated the expression of Keap 1 and Cox2(P<0.05).Conclusion:Wenfei Jiangzhuo Formula may inhibit ferroptosis by regulating the expression of proteins associated with the Nrf2/Keap 1-GPx4 signaling pathway to reduce the deposition of iron particles in the hippocampus,thus improving the cognitive condition of the brain.Wenfei Jiangzhuo Formula administrated at 11.8 g/kg demonstrated the best effect.

关 键 词：温肺降浊方 核因子-红细胞2相关因子2/Kelch样ECH关联蛋白1/谷胱甘肽过氧化物酶4信号通路 铁死亡 血管性痴呆 认知功能障碍 

分 类 号：R285.5[医药卫生—中药学]