Atractylenolide-1 Attenuates TNF-?-induced Apoptosis And Autophaghy In Mouse C2C12 Myoblasts

BACKGROUNDSkeletal muscle is composed of muscle cells(fibers),accounts for about 50%of total body quality,the body of any activity is performed by the contraction of skeletal muscles,it directly affects the strength and endurance of the human body.Skeletal muscle atrophy refers to the quality and function of the skeletal muscle loss(power and performance),physiological conditions such as old age can appear the situation of skeletal muscle atrophy,pathological state of all kinds of chronic wasting diseases such as chronic renal failure,chronic heart failure,chronic obstructive pulmonary disease(COPD),leukemia,cancer,diabetes,AIDS,autoimmune diseases and so on all can lead to skeletal muscle atrophy.Study founds that tumor necrosis factor-a(TNF-a)plays an important role in the pathological process of skeletal muscle atrophy,such as influences muscle cell growth,differentiation,apoptosis and necrosis.previous research smggests that TNF-a pathological rise is closely related to skeletal muscle atrophy,it can inhibit the growth of muscle cells,proliferation,differentiation,and induced muscle cell apoptosis and autophagy.Cells apoptosis is a autonomic ordered cell death for the sake of maintaining the stability of milieu interne,autonomic ordered by the genes that control cell death,cell apoptosis pathways including internal mitochondrial pathway and death receptor mediated external apoptosis pathway and endoplasmic reticulum stress pathway.To maintain the stability of internal environment of the body and growth plays an important role,but the excessive apoptosis can lead to muscle atrophy.Study confirmed that autophagy is closely related to the skeletal muscle atrophy,in skeletal muscle tissue,autophagy is the primary way to catabolism,its degradation of storage proteins in cells,directly induce skeletal muscle protein catabolism,autophagy can also react on mTOR influence anabolic signal,these processes are associated with skeletal muscle atrophy.Atractylenolide-1 is isolated from volatile oil components from Chinese herbal medicine atractylodes,has obvious antioxidant,anti-aging,alzheimer's disease,antitumor,anti-inflammatory and immune regulation,and so on.But its in skeletal muscle apoptosis and autophagy in the research,there is no reported this study aims to explore the lactone 1 limit of TNF alpha induced C2C12 mice skeletal muscle cells apoptosis and inhibition of autophagy and the underlying mechanisms,for the treatment of diseases in skeletal muscle atrophy associated provide new theoretical basis.OBJECTIVEThe purpose of the research was to appraise the protective effection along with the underlying mechanisms by which acts on TNF-a-induced apoptosis and autophagy in mouse C2C12 myoblasts.METHODSC2C12 cells as the research object,Atractylenolide-1 separately toxicity tests to determine the appropriate concentration of drmg intervention,intervention group(normal control group,20 ng/mlTNF-?model group,the TNF-? + Atractylenolide-1 separately different dose groups(10,20,30?g/mL)),respectively Hoechst33342 colouring be applied to appraise apoptosis rate,flow cytometry to detect apoptotic cells,mitochondrial membrane potential detection kits JC-1 cell mitochondrial membrane potential,fluorescence microscope cell autophagy,protein immunoblot method to detect apoptosis and autophagy related proteins.RESULTS1.The method of CCK 8 concentration for 40?g/mL detection results indicate atractylodes lactone 1 significantly inhibit the growth of C2C12 skeletal muscle cells,the discrepancy was conspicuous in statistical(P<0.05),the concentration under 30 ?g/mL of atractylodes lactone 1 not obvious effects on cells.2.Compared with DMSO control group,the TNF-amodel group significantly increased level of reactive oxygen species,atractylodes lactone 1 different dose groups(10,20,30?g/mL))intervention group active oxygen levels decreased significantly,the discrepancy was conspicuous in statistical(P<0.05).3.Atractylenolide-1 inhibits TNF-a-induced C2C12 skeletal muscle cells apoptosis by inhibiting ASK1 JNK/p38 lightning signaling pathways.4.Atractylenolide-1 inhibits TNF-a-induced C2C12 skeletal muscle cells autophagy with invokeing the phosphorylated protein kinase signaling pathways.CONCLUSIONAtractylenolidel can inhibits TNF-a-induced C2C12 skeletal muscle cells apoptosis and autophagy by inhibiting ASK1 JNK/p38 signaling pathways as well as activating the Akt signaling pathways.