茱萸丸调控p53/SLC7A11信号通路介导氧化损伤及铁死亡减轻动脉粥样硬化  

Zhuyu Pills regulate p53/SLC7A11 signaling pathway-mediated oxidative damage and ferroptosis to treat atherosclerosis

作　　者：宋玮 张钟艺 张小波 沈涛[1] SONG Wei;ZHANG Zhong-yi;ZHANG Xiao-bo;SHEN Tao(Chengdu University of Traditional Chinese Medicine,Chengdu 610075,China)

机构地区：[1]成都中医药大学,四川成都610075

出　　处：《中国中药杂志》2024年第15期4118-4127,共10页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81973743,82374331)。

摘　　要：旨在探讨茱萸丸对动脉粥样硬化(AS)的影响及可能的作用机制。采用高脂饲料喂养诱导小鼠AS模型,造模周期为12周。造模成功的50只载脂蛋白E基因敲除(ApoE^(-/-))小鼠按照随机数字表法分为5组,即模型组,茱萸丸低、中、高剂量组,阿托伐他汀钙组,每组10只;C57BL/6J小鼠10只作为空白组。空白组及模型组给予等体积无菌蒸馏水灌胃,茱萸丸低、中、高剂量组给予130.54、261.08、522.16 mg·kg^(-1)灌胃,阿托伐他汀钙组给予10.40 mg·kg^(-1)灌胃,每日1次,共12周。给药结束后,采用苏木素-伊红(HE)染色观察小鼠主动脉、肝脏、附睾脂肪的病理学变化,并计算主动脉斑块面积占比、附睾脂肪面积、非酒精性脂肪肝活动性积分(NAS);油红O染色、Masson染色观察主动脉脂质及胶原纤维沉积,并计算主动脉红染脂质面积占比、主动脉胶原沉积面积占比;比色法检测血清超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)、铁离子水平;免疫荧光法检测主动脉环氧合酶2(COX2)、铁蛋白重链1(FTH1)、胱氨酸/谷氨酸逆向转运蛋白溶质载体家族7成员11(SLC7A11)、谷胱甘肽过氧化物酶4(GPX4)蛋白水平;免疫组化法检测主动脉肿瘤蛋白53(p53)水平;蛋白免疫印迹法检测主动脉p53、SLC7A11蛋白表达;实时荧光定量聚合酶链式反应(real-time PCR)检测小鼠主动脉p53、SLC7A11、GPX4、FTH1、前列腺素G/H合酶2(PTGS2)、还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶1(NOX1)mRNA表达水平。结果显示,与空白组比较,模型组小鼠主动脉斑块面积明显扩大,胶原纤维沉积增加,肝脏脂质沉积增加,脂滴变多,附睾脂肪细胞体积扩大;血清中铁离子、MDA含量升高,SOD、GSH-Px水平降低;p53、COX2蛋白表达升高;主动脉FTH1、SLC7A11、GPX4蛋白及mRNA表达降低;PTGS2、NOX1 mRNA表达升高。与模型组比较,茱萸丸低、中、高剂量组及阿托伐他汀钙组小鼠主动脉斑�This article aims to investigate the effect of Zhuyu Pills on atherosclerosis(AS)and decipher the underlying mechanism.The mouse model of AS was established by feeding with a high-fat diet for 12 weeks.The 50 successfully modeled mice with the apolipoprotein E knockout(ApoE^(-/-))were assigned by the random number table method into 5 groups(n=10):model,low-,medium-,and high-dose(130.54,261.08,522.16 mg·kg^(-1),respectively)Zhuyu Pills,and atorvastatin calcium(10.40 mg·kg^(-1)).Ten C57BL/6J mice were selected as the blank group.The blank group and model group were administrated with an equal volume of normal saline,and other groups were administrated with corresponding drugs once a day for 12 weeks.At the end of drug intervention,hematoxylin-eosin(HE)staining was employed to observe the pathological changes of fat in the aorta,liver,and epididymis of mice,and the proportion of aortic plaque area,fat area in epididymis,and nonalcoholic fatty liver disease activity score(NAS)were calculated.Lipid and collagen deposition in the aorta was observed by oil red O staining and Masson staining,respectively,and the proportions of lipid and collagen deposition areas were calculated.The serum levels of superoxide dismutase(SOD),malondialdehyde(MDA),glutathione peroxidase(GSH-Px),and iron ion were measured by colorimetry.The expression of cyclooxygenase 2(COX2),ferritin heavy chain 1(FTH1),cystine/glutamate reverse transporter solute carrier family 7 member 11(SLC7A11),and glutathione peroxidase 4(GPX4)in the aorta was detected by the immunofluorescence assay.The level of tumor protein 53(p53)in the aorta was detected by immunohistochemistry.The protein levels of p53 and SLC7A11 in the aorta were determined by Western blot.The mRNA levels of p53,SLC7A11,GPX4,FTH1,prostaglandin G/H synthase 2(PTGS2),and reduced nicotinamide adenine dinucleotide phosphate oxidase 1(NOX1)in mouse aorta were determined by real-time PCR.The results showed that compared with the blank group,the model group showcased enlarged aortic plaque area,inc

关 键 词：动脉粥样硬化 茱萸丸 氧化损伤 铁死亡 肿瘤蛋白53(p53) 胱氨酸/谷氨酸逆向转运蛋白溶质载体家族7成员11(SLC7A11) 

分 类 号：R285.5[医药卫生—中药学]