摘要
目的探讨双和汤通过调控肌肉铁死亡改善小鼠肌少症的效应和机制。方法老龄SAMP8小鼠随机分为对照组(生理盐水)、双和汤组[双和汤,16 g/(kg·d)]和铁死亡抑制剂Ferrostatin-1(Ferr-1)腹腔注射组[Ferr-1组,0.7 mg/(kg·d)],每组15只。灌胃干预12周,注射干预3周。干预结束后测量小鼠各部分肌肉质量,肌肉测试系统检测小鼠肌肉力量,握力仪检测小鼠最大抓握力,Laminin染色检测小鼠肌纤维显微结构,MHC染色检测小鼠肌纤维类型,还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)、还原型谷胱甘肽(GSH)、脂质氧化丙二醛(MDA)含量检测及4-HNE染色检测小鼠肌肉组织脂质氧化程度,Western Blot检测小鼠肌肉组织铁死亡标志蛋白谷胱甘肽过氧化物酶4(GPX4)、溶质载体家族7成员11(SLC7A11)的表达,透射电镜检测小鼠肌纤维线粒体形态及分布,Western Blot及免疫荧光染色检测小鼠肌肉组织铁调节蛋白2/双微体同源基因2/肿瘤蛋白P53(IRP2/MDM2/P53)信号通路的改变。结果与对照组比较,双和汤组肌少症小鼠各部分肌肉含量提高(P<0.01),肌肉力量和抓握力增强(P<0.01),肌纤维萎缩形态、老化亚型和线粒体形态改善(P<0.01),肌肉组织IRP2/MDM2/P53信号改善(P<0.01),NAPDH和GSH含量升高(P<0.01),MDA和4-HNE水平降低(P<0.01),铁死亡标志蛋白GPX4和SLC7A11表达增加(P<0.01)。结论双和汤通过调控IRP2/MDM2/P53信号通路,减少肌肉铁死亡,从而有效改善老龄小鼠肌少症症状。
Objective To investigate the effects and mechanisms of Shuanghe Decoction on sarcopenia by attenuating ferroptosis of muscles in mice.Methods Aged SAMP8 mice were randomly divided into saline gavage group(control group),Shuanghe Decoction group(Shuanghe group,16 g·kg-1·d-1),and Ferrostatin-1(Ferr-1)intraperitoneal injection group(Ferr-1 group,0.7 mg·kg-1·d-1).The gavage intervention lasted 12 weeks and the injection intervention was administered for 3 weeks.After intervention,the muscle mass of various parts was measured,muscle strength was detected by muscle testing system,maximum grip strength was detected by grip dynamometer,microstructure of mouse muscle fibers was detected by Laminin staining,muscle fiber type was detected by MHC staining,lipid oxidation of mouse muscle tissues was detected by NAPDH,reduced glutathione(GSH),and malonaldehyde(MDA)content detection and 4-HNE staining,and the expression of GPX4 and SLC7A11 as ferroptosis markers in muscle tissues was detected by Western Blot.Transmission electron microscopy was used to detect the mitochondrial morphology and distribution of mouse muscle fibers,and Western Blot and immunofluorescence staining was used to detect the alteration of the IRP2/MDM2/P53 signaling axis in mouse muscle tissues.Results Compared with the control group,the muscle mass of sarcopenia mice of Shuanghe group increased(P<0.01),the muscle strength and grip strength enhanced(P<0.01),the muscle fiber atrophy morphology,MHC isoforms and mitochondrial morphology improved(P<0.01),IRP2/MDM2/P53 signaling was modulated(P<0.01),the levels of NADPH and GSH increased(P<0.01),lipid peroxidation reduced(P<0.01),MDA and 4-HNE levels decreased(P<0.01),and the expression GPX4 and SLC7A11 as ferroptosis marker proteins in muscles upregulated(P<0.01).Conclusion Shuanghe Decoction can effectively ameliorate sarcopenia in aged mice by attenuating ferroptosis in muscles through the IRP2/MDM2/P53 pathway.
作者
黄研
齐真
陈川
邢三丽
申定珠
HUANG Yan;QI Zhen;CHEN Chuan;XING San-li;SHEN Ding-zhu(Shanghai Geriatric Institute of Chinese Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai,200031)
出处
《中国中西医结合杂志》
北大核心
2025年第11期1365-1372,共8页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家自然科学基金资助项目(No.82305352)。
关键词
肌少症
双和汤
SAMP8小鼠
铁死亡
中药复方
sarcopenia
Shuanghe Decoction
senescence accelerated mouse prone 8
ferroptosis
Chinese herbal medicine compound prescription
