补肾还精方通过调节铁死亡干预小鼠肌少症的效应和机制研究  

Effect and mechanism of Bushen Huanjing Recipe on sarcopenia mice by regulating ferroptosis

作　　者：黄研 邢三丽[1] 胡怡然 陈川[1] HUANG Yan;XING Sanli;HU Yiran;CHEN Chuan(Shanghai Geriatric Institute of Chinese Medicine,Shanghai 200031,China;Shanghai University of Traditional Chinese Medicine,Shanghai201203,China)

机构地区：[1]上海市中医老年医学研究所,上海200031 [2]上海中医药大学,上海201203

出　　处：《上海中医药杂志》2022年第7期74-82,共9页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81774019);上海市自然科学基金项目(20ZR1453500)。

摘　　要：目的 探讨补肾还精方干预小鼠肌少症的效应和机制。方法 采用老龄快速老化小鼠品系8(SAMP8)小鼠作为肌少症模型,随机分为对照组(0.9%NaCl溶液灌胃组)、补肾组(补肾还精方灌胃组)和Ferr-1组(铁死亡抑制剂Ferrostatin-1腹腔注射组)。补肾组和对照组灌胃干预12周,Ferr-1组干预3周。干预结束后测量小鼠各部分肌肉含量;肌肉测试系统检测小鼠肌肉力量;握力仪检测小鼠抓握力;层粘连蛋白免疫荧光染色检测小鼠肌肉纤维显微结构;检测还原型烟酰胺腺嘌呤二核苷酸磷酸(NAPDH)、谷胱甘肽(GSH)、脂质氧化指标丙二醛(MDA)含量及免疫组化染色检测4-羟基壬烯醛(4-HNE)含量,观察小鼠肌肉组织脂质氧化程度;Western blot检测小鼠肌肉组织中铁死亡调控蛋白Gpx4、SCL7A11、P53及P21的表达情况;透射电镜检测小鼠肌肉纤维线粒体形态。结果 与对照组比较,补肾组、Ferr-1组小鼠各部分肌肉含量提高,肌肉力量和抓握能力提升,肌肉纤维面积增大,肌肉组织脂质过氧化程度降低(P<0.01)。与对照组比较,补肾组、Ferr-1组小鼠肌肉组织铁死亡的两个负相关标志蛋白谷胱甘肽过氧化物酶4(Gpx4)和溶质载体家族7成员11(SLC7A11)的表达显著增加(P<0.001);而负调控SLC7A11的细胞衰老相关蛋白P53及其下游蛋白P21的表达却显著减少(P<0.001),肌纤维线粒体铁死亡形态改善(P<0.01)。结论 补肾还精方可通过调控肌肉组织铁死亡途径,有效干预治疗小鼠肌少症。Objective accelerated mouse prone 8(SAMP8)was used as a sarcopenia model,and 36 SAMP8 mice were randomly divided into 0.9%NaCl solution group(control group),Bushen Huanjing Recipe group(Bushen group)and ferroptosis inhibitor Ferrostatin-1(Ferr-1)intraperitoneal injection group(Ferr-1 group). Gavage intervention was given in the control and the Bushen groups for 12 weeks,and inhibitor injection intervention was given for 3 weeks.After the intervention,the muscle mass of the mice was weighed. The muscle strength of the mice was detected by a muscle functional test system,and the maximum grip strength of the mice was detected by a grip strength meter. The microstructure of the mice muscle fibers was detected by the laminin immunofluorescence staining. The contents of reduced nicotinamide adenine dinucleotide phosphate(NAPDH), glutathione(GSH),malondialdehyde(MDA)and 4-hydroxynonenal(4-HNE)by immunohistochemistry staining were used to detect lipid oxidation in muscles of the mice. The expression of ferroptosis related proteins such as anti-glutathione peroxidase 4(Gpx4),SCL7A11,P53 and P21 in mice muscles was detected by Western blot,and the morphology of mitochondria of muscle fiber in the mice were detected by a transmission electron microscope.ResultsCompared with the control group,Bushen Huanjing Recipe and Ferr-1 intervention increased the muscle mass,strength and grip strength as well as the area of muscle fibers in the mice,improved the morphology of mitochondria in muscle fibers(P<0.01),and reduced lipid peroxidation. The expression of antioxidant glutathione peroxidase(Gpx4)and SLC7A11,the key element of cystine/glutamate antiporter,was significantly increased(P< 0.001),while the expression of cell senescence-related protein p53 and its downstream protein P21,which negatively regulated SLC7A11,was significantly decreased(P<0.001).ConclusionThe Bushen Huanjing Recipe can effectively treat sarcopenia in mice by regulating ferroptosis.

关 键 词：肌少症 补肾还精方 小鼠模型 铁死亡 中药研究 

分 类 号：R285.5[医药卫生—中药学]