健脾方对废用性肌肉萎缩小鼠Akt/FOXO3a通路表达的机制研究  

Mechanism of Jianpi Formula on expression of Akt/FOXO3a pathway in disused muscle atrophy mice

作　　者：李小凯 丁劼 孙雯[1,2] 高洒洒 杨光月 郭海玲[1,2] 赵咏芳[1,2] LI Xiaokai;DING Jie;SUN Wen;GAO Sasa;YANG Guangyue;GUO Hailing;ZHAO Yongfang(Shi's Center of Orthopedics and Traumatology,Shuguang Hospital Affliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Institute of Traumatology&Orthopedics,Shanghai Academy of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属曙光医院石氏伤科医学中心,上海201203 [2]上海市中医药研究院骨伤科研究所,上海201203

出　　处：《上海中医药大学学报》2022年第5期86-92,共7页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(82174404、81774342);上海市科委医学创新研究专项重大项目(21Y31920200);上海市科委项目(17401901900);上海市慢性筋骨病临床医学研究中心建设项目(20MC1920600);上海市临床重点专科“中医骨伤科”建设项目(shslczdzk03901);全国中医学术流派传承工作室“石氏伤科流派传承工作室”建设项目;上海申康医院发展中心临床三年行动计划资助项目(SHDC2020CR3090B)。

摘　　要：目的:探究健脾方基于Akt/FOXO3a信号通路预防小鼠废用性肌肉萎缩的作用机制。方法:将45只10周龄雄性C57/BL6小鼠随机分为对照组、模型组及健脾方组。通过后肢卸载构建废用性肌肉萎缩小鼠模型。对照组和模型组小鼠每天每只灌胃给予0.2 ml 0.9%NaCl溶液,健脾方组小鼠每天每只灌胃给予0.2 ml健脾方混悬液。给药28 d后,称取小鼠体质量、肌肉湿重(累积肌肉、腓肠肌、比目鱼肌、胫骨前肌、趾长伸肌及股四头肌),qRT-PCR检测腓肠肌中IGF-1、PI3K、Akt、Atrogin及MuRF1 mRNA的表达水平,Western blot检测腓肠肌中p-Akt、p-FOXO3a、Akt、FOXO3a蛋白的表达水平。结果:(1)与对照组比较,模型组小鼠四肢抓力/体质量比值、肌肉湿重/体质量比值、肌纤维横截面积和直径明显降低(P<0.05)。与模型组比较,健脾方组小鼠四肢抓力/体质量比值明显增加(P<0.05);累积肌肉、比目鱼肌、胫骨前肌、股四头肌的湿重/体质量比值明显增加(P<0.05);肌纤维横截面积明显增加(P<0.05)。(2)健脾方干预后,小鼠腓肠肌IGF-1、Akt mRNA表达明显增加(P<0.05),Atrogin、MuRF1 mRNA表达明显减少(P<0.05);p-Akt蛋白表达升高,p-FOXO3a蛋白表达降低。结论:健脾方可能通过上调Akt磷酸化水平抑制Akt/FOXO3a信号通路来预防废用性肌肉萎缩。Objective:To explore the mechanism of Jianpi Formula in preventing disuse muscle atrophy in mice based on Akt/FOXO3a signaling pathway.Methods:Forty-five male C57/BL6 mice were randomly divided into control group,model group and Jianpi Formula group. Disuse muscular atrophy mouse model was established by hindlimb unloading. Mice in control group and model group were given 0.2 ml 0.9% NaCl solution by gavage every day,and mice in Jianpi Formula group were given 0.2 ml Jianpi Formula suspension by gavage every day. After 28 d of drug administration,the body weight and wet muscle weight(accumulation of muscle,gastrocnemius muscle,soleus muscle,tibialis anterior muscle,extensor digitorum longus muscle and quadriceps femoris muscle)of mice were weighed.Moreover,qRT-PCR was used to detect IGF-1,PI3K,Akt,Atrogin and MuRF1 mRNA expression level,Western blot was used to detect p-Akt,p-FOXO3a,Akt and FOXO3a protein expression level in gastrocnemius muscle.Results:(1) Compared with the control group,the grip strength/body weight ratio,wet weight/body weight ratio of muscles,cross-sectional area and diameter of muscle fiber in mice of the model group significantly decreased(P<0.05). Compared with the model group,the grip strength/body weight ratio significantly increased(P<0.05);the wet weight/body weight ratio of cumulative muscle,soleus muscle,tibialis anterior muscle and quadriceps femoris muscle significantly increased(P<0.05);muscle fiber cross-sectional area significantly increased(P<0.05).(2)After the intervention of Jianpi Formula,IGF-1 and Akt mRNA expression significantly increased(P<0.05)in gastrocnemius muscle of mice,while Atrogin and MuRF1 mRNA expression significantly decreased(P<0.05);p-Akt protein expression increased,p-FOXO3a protein expression decreased.Conclusion :Jianpi Formula may be prevent disuse muscular atrophy by inhibiting Akt/FOXO3a signaling pathway through upregulating Akt phosphorylation.

关 键 词：健脾方 废用性肌萎缩 后肢卸载 抓力 腓肠肌 Akt/FOXO3a信号通路 

分 类 号：R285.5[医药卫生—中药学]