补肺健脾方对COPD大鼠骨骼肌IGF-1、mTOR和HIF1-α的影响
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摘要
目的:观察补肺健脾方对慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)稳定期模型大鼠股四头肌、肱二头肌、肋间肌、比目鱼肌胰岛素样生长因子(insulin-like growth factor,IGF)~(-1)、哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)和低氧诱导因子(hypoxia-inducible factor,HIF)~(-1)α表达的影响。方法:48只SD大鼠随机分为空白组、模型组、氨茶碱组、补肺健脾组,每组12只。采用香烟烟雾暴露联合细菌感染法制备COPD稳定期大鼠模型。自第9周起,对照组、模型组给予2 mL生理盐水灌胃;补肺健脾组、氨茶碱组给予补肺健脾方[5 g·(kg·d)~(-1)]、氨茶碱[2.3 m g·(kg·d)~(-1)]治疗,第20周结束后取材。采用荧光定量PCR和Western Blot技术观察4种骨骼肌IGF-1、mTOR和HIF1-αmRNA和蛋白表达。结果:与对照组比较,模型组IGF-1、mTOR mRNA及蛋白表达显著降低(P<0.05),HIF1-αmRNA及蛋白表达显著升高(P<0.01)。与模型组比较,补肺健脾组和氨茶碱组IGF-1、mTOR mRNA表达显著升高(P<0.01),IGF-1、mTOR蛋白表达升高(P<0.05),HIF1-αmRNA和蛋白表达显著降低(P<0.01)。结论:补肺健脾方可通过显著上调IGF-1、mTOR,下调HIF1-α,改善COPD大鼠骨骼肌功能。
Objective:To observe the effects of Bufei Jianpi Formula on the expressions of insulin-like growth factor(IGF)~(-1),mammalian target of rapamycin(mTOR) and hypoxia-inducible factor(HIF)~(-1)α in quadriceps femoris,biceps brachii,intercostal muscle and soleus muscles in rats with chronic obstructive pulmonary disease(COPD).Methods:SD rats were randomly divided into control,model,aminophylline and Bufei Jianpi groups.12 rats in each group.COPD rat model was prepared by cigarette smoke exposure combined with bacterial infections.Rats in control and model groups were given 2 mL of saline,while rats in the rest two groups were administrated with Bufei Jianpi Formula [5 g·(kg·d)~(-1)]or aminophylline [2.3 mg·(kg·d)~(-1)],respectively,from week 9 up to 20.Muscle tissues were harvested at week 20.The expressions of IGF-1,mTOR and HIF1-α mRNA and protein in four muscles were observed by using quantitative PCR and Western Blot techniques.Results:IGF-1 and mTOR mRNA and protein in the four muscle were significantly lower in model group than control(P<0.05),while HIF1-α was significantly higher(P<0.01).Compared with model group,IGF-1 and mTOR mRNA and protein was significantly increased in Bufei Jianpi and aminophylline group(P<0.01 or P<0.05),while HIF1-α mRNA and protein expression were significantly decreased(P<0.01).Conclusion:Bufei Jianpi Formula can improve skeletal muscle function via up-regulating the expressions of IGF-1,mTOR and depressing HIF1-α.
Objective:To observe the effects of Bufei Jianpi Formula on the expressions of insulin-like growth factor(IGF)~(-1),mammalian target of rapamycin(mTOR) and hypoxia-inducible factor(HIF)~(-1)α in quadriceps femoris,biceps brachii,intercostal muscle and soleus muscles in rats with chronic obstructive pulmonary disease(COPD).Methods:SD rats were randomly divided into control,model,aminophylline and Bufei Jianpi groups.12 rats in each group.COPD rat model was prepared by cigarette smoke exposure combined with bacterial infections.Rats in control and model groups were given 2 mL of saline,while rats in the rest two groups were administrated with Bufei Jianpi Formula [5 g·(kg·d)~(-1)]or aminophylline [2.3 mg·(kg·d)~(-1)],respectively,from week 9 up to 20.Muscle tissues were harvested at week 20.The expressions of IGF-1,mTOR and HIF1-α mRNA and protein in four muscles were observed by using quantitative PCR and Western Blot techniques.Results:IGF-1 and mTOR mRNA and protein in the four muscle were significantly lower in model group than control(P<0.05),while HIF1-α was significantly higher(P<0.01).Compared with model group,IGF-1 and mTOR mRNA and protein was significantly increased in Bufei Jianpi and aminophylline group(P<0.01 or P<0.05),while HIF1-α mRNA and protein expression were significantly decreased(P<0.01).Conclusion:Bufei Jianpi Formula can improve skeletal muscle function via up-regulating the expressions of IGF-1,mTOR and depressing HIF1-α.
引文
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[12]毛静,王丽丽,李亚,等.补肺健脾方对COPD大鼠骨骼肌Fas、Fasl、Bax及Bcl-2表达的影响[J].中华中医药杂志,2017,32(2):746-749.
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[14]李素云,李亚,李建生,等.补肺健脾方对慢性阻塞性肺疾病大鼠膈神经放电和膈肌功能的影响[J].中国中西医结合杂志,2012,32(6):812-816.
[15]LI S Y,LI J S,WANG M H,et al.Effects of comprehensive therapy based on traditional Chinese medicine patterns in stable chronic obstructive pulmonary disease:a four-center,openlabel,randomized,controlled study[J].BMC Complement Altern Med,2012,12:197.
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[17]HOPPELER H,VOGT M,WEIBEL E R,et al.Response of skeletal muscle mitochondria to hypoxia[J].Exp Physiol,2003,88(1):109-119.
[18]万焕真.胰岛素样生长因子对合成代谢和免疫功能的影响[J].国外医学·骨科学分册,1996,17(4):207-210.
[19]COSKUN F,EGE E,UZASLAN E,et al.Evaluation of thyroid hormone levels and somatomedin-C(IGF-1)in patients with chronic obstructive pulmonary disease(COPD)and relation with theseverity of the disease[J].Tuberk Toraks,2009,57(4):369-375.
[20]孙丽华,欧阳晓平,谷伟,等.胰岛素样生长因子对COPD大鼠膈肌凋亡的保护作用及对肺功能的影响[J].中国呼吸与危重监护杂志,2009,8(5):441-445.
[21]段红杰.胰岛素样生长因子-I抗骨骼肌萎缩研究进展[J].军医进修学院学报,2009,30(1):109-110.
[22]XU G,ZHANG W,BETRAM P,et al.Pharmacogenomic profiling of the PI3K/PTEN-AKT-m TOR pathway in common human tumor[J].Int J Oncol,2004,24(4):893-900.
[23]刘娟.哺乳动物雷帕霉素靶蛋白信号通路在吸烟诱导的气道炎症和慢性阻塞性肺疾病中的调控作用及机制研究[D].杭州:浙江大学,2015.
[24]李艳利,韩锋锋,李妍.PI3K/AKT/m TOR途径在慢性阻塞性肺疾病大鼠骨骼肌萎缩中的作用[J].中国呼吸与危重监护杂志,2014,13(5):474-479.
[25]魏文婷,李冰,贾晔然,等.慢性阻塞性肺疾病患者肺组织中缺氧诱导因子1α的表达变化[J].中国病理生理杂志,2013,29(9):1700-1703.
[26]李启芳,戴爱国,徐平.慢性阻塞性肺疾病患者肺小血管低氧诱导因子1α的表达[J].中华内科杂志,2006,45(2):136-139.
[2]GBD 2013 Mortality and Causes of Death Collaborators.Global,regional,and national age-sex specific all-cause and cause-specific mortality for 240 causes of death,1990-2013:a systematic analysis for the Global Burden of Disease Study 2013[J].Lancet,2015,385(9963):117-171.
[3]KELLY J L,ELKIN S L,FLUXMAN J,et al.Breathlessness and skeletal muscle weakness in patients undergoing lung health screening in primary care[J].COPD,2013,10(1):40-54.
[4]DONG Y,LI Y,SUN Y,et al.Bufei Jianpi granules improve skeletal muscle and mitochondrial dysfunction in rats with chronic obstructive pulmonary disease[J].BMC Complement Altern Med,2015,15:51.
[5]VAN DEN BORST B,GOSKER H R,SCHOLS A M.Central fat and peripheral muscle:partners in crime in chronic obstructive pulmonary disease[J].Am J Respir Crit Care Med,2013,187(1):8-13.
[6]SHRIKRISHNA D,PATEL M,TANNER R J,et al.Quadriceps wasting and physical inactivity in patients with COPD[J].Eur Respir J,2012,40(5):1115-1122.
[7]李江.胰岛素样生长因子-1的研究进展[J].国际检验医学杂志,2013,34(7):848-850.
[8]黄娇.低氧微环境对骨髓间充质干细胞成骨分化影响的研究[D].重庆:重庆医科大学,2012.
[9]李启荣,师永红.缺氧诱导因子1降解-泛素化研究进展[J].中华肿瘤防治杂志,2013,20(17):1371-1374.
[10]徐益文,徐庆祥,丁义涛.缺氧诱导因子1与肿瘤的研究进展[J].医学综述,2012,18(11):1656-1660.
[11]MALTAIS F,DECRAMER M,CASABURI R,et al.An official American Thoracic Society/European Respiratory Society statement:update on limb muscle dysfunction in chronic obstructive pulmonary disease[J].Am J Respir Crit Care Med,2014,189(9):15-62.
[12]毛静,王丽丽,李亚,等.补肺健脾方对COPD大鼠骨骼肌Fas、Fasl、Bax及Bcl-2表达的影响[J].中华中医药杂志,2017,32(2):746-749.
[13]LI Y,LI S Y,LI J S,et al.A rat model for stable chronic obstructive pulmonary disease induced by cigarette smoke inhalation and repetitive bacterial infection[J].Biol Pharm Bull,2012,35(10):1752-1760.
[14]李素云,李亚,李建生,等.补肺健脾方对慢性阻塞性肺疾病大鼠膈神经放电和膈肌功能的影响[J].中国中西医结合杂志,2012,32(6):812-816.
[15]LI S Y,LI J S,WANG M H,et al.Effects of comprehensive therapy based on traditional Chinese medicine patterns in stable chronic obstructive pulmonary disease:a four-center,openlabel,randomized,controlled study[J].BMC Complement Altern Med,2012,12:197.
[16]RAGUSO C A,GUINOT S L,JANSSENS J P,et al.Chronic hypoxia:commontraits between chronic obstructive pulmonary disease and altitude[J].Curr Opin Clin Nutr Metab Care,2004,7(4):411-417.
[17]HOPPELER H,VOGT M,WEIBEL E R,et al.Response of skeletal muscle mitochondria to hypoxia[J].Exp Physiol,2003,88(1):109-119.
[18]万焕真.胰岛素样生长因子对合成代谢和免疫功能的影响[J].国外医学·骨科学分册,1996,17(4):207-210.
[19]COSKUN F,EGE E,UZASLAN E,et al.Evaluation of thyroid hormone levels and somatomedin-C(IGF-1)in patients with chronic obstructive pulmonary disease(COPD)and relation with theseverity of the disease[J].Tuberk Toraks,2009,57(4):369-375.
[20]孙丽华,欧阳晓平,谷伟,等.胰岛素样生长因子对COPD大鼠膈肌凋亡的保护作用及对肺功能的影响[J].中国呼吸与危重监护杂志,2009,8(5):441-445.
[21]段红杰.胰岛素样生长因子-I抗骨骼肌萎缩研究进展[J].军医进修学院学报,2009,30(1):109-110.
[22]XU G,ZHANG W,BETRAM P,et al.Pharmacogenomic profiling of the PI3K/PTEN-AKT-m TOR pathway in common human tumor[J].Int J Oncol,2004,24(4):893-900.
[23]刘娟.哺乳动物雷帕霉素靶蛋白信号通路在吸烟诱导的气道炎症和慢性阻塞性肺疾病中的调控作用及机制研究[D].杭州:浙江大学,2015.
[24]李艳利,韩锋锋,李妍.PI3K/AKT/m TOR途径在慢性阻塞性肺疾病大鼠骨骼肌萎缩中的作用[J].中国呼吸与危重监护杂志,2014,13(5):474-479.
[25]魏文婷,李冰,贾晔然,等.慢性阻塞性肺疾病患者肺组织中缺氧诱导因子1α的表达变化[J].中国病理生理杂志,2013,29(9):1700-1703.
[26]李启芳,戴爱国,徐平.慢性阻塞性肺疾病患者肺小血管低氧诱导因子1α的表达[J].中华内科杂志,2006,45(2):136-139.