过氧化氢对成肌细胞的氧化损伤作用研究

doi:10.11843/j.issn.0366-6964.2019.05.012

Abstract

Abstract:

The change of C2C12 cells exposed to H₂O₂ was investigated and the possible path of oxidative damage was explored in this study. There was 3 treatments, and 6 repeats in each treatment. C2C12 cells attached to the substrate and cultured in DMEM for 24 h were treated as control (n=6); the medium was added with PDTC (20 μmol·L^-1, PDTC treatment, n=6) for 24 h or H₂O₂ (0.5 mmol·L^-1, H₂O₂ treatment, n=6) for 24 h. The viability, ROS level and apoptosis rate of cells in different treatments were detected. The expression of genes related to cell apoptosis and autophagy and factors related to NF-κB signaling pathway were determined by real-time PCR and Western blot. The results showed that cells in PDTC treatment had significantly higher total apoptosis rate, mRNA expression of caspase-3, caspase-6, caspase-9 and the protein expression of total caspase-3, mRNA expression of Beclin1, LC3-Ⅱ/LC3-I and the protein expression level of total LC3-Ⅱ(P<0.05), and had significantly lower mRNA expression of p50, Bcl-2, RelA, Cox-2 and protein expression of total NF-κB compared to that in the control treatment(P<0.05). Moreover, cells in H₂O₂ treatment exhibited significantly higher ROS level and total apoptosis rate(P<0.05), mRNA expression of caspase-3, caspase-6, caspase-8, caspase-9 and protein expression of total caspase-3(P<0.05), mRNA expression of Beclin 1, LC3-Ⅱ/LC3-I and protein expression of total LC3-Ⅱ(P<0.05), mRNA expression of Bax(P<0.05), and had significantly lower mRNA expressions of p50, Bcl-2, RelA, Cox-1, Cox-2 and protein expression of total NF-κB(P<0.05) compared to that in the control treatment. These results indicated that H₂O₂ could promote the ROS formation, mediated the apoptosis and autophagy processes by suppressing the NF-κB signaling pathway in C2C12 cells, which was similar with the role of PDTC, the inhibitor of NF-κB factor.

CLC Number:

S852.2

CHEN Xiangxing, LI Jiaolong, XING Tong, ZHANG Lin, GAO Feng. Effects of H₂O₂ on Oxidative Damage of C2C12 Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(5): 1016-1025.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.xmsyxb.com/EN/10.11843/j.issn.0366-6964.2019.05.012

https://www.xmsyxb.com/EN/Y2019/V50/I5/1016

References

[1] 赖桂华,邱小忠,余磊,等.氧化应激对C2C12成肌细胞增殖与凋亡的影响[J].中国组织工程研究与临床康复,2007,11(2):278-281.
LAI G H,QIU X Z,YU L,et al.Effect of oxidative stress on myoblast C2C12 proliferation and apoptosis[J].Journal of Clinical Rehabilitative Tissue Engineering Research,2007,11(2):278-281.(in Chinese)
[2] 邱小忠,余磊,廖华,等.活性氧促使L6成肌细胞的分化[J].第一军医大学学报,2005,25(11):1384-1386.
QIU X Z,YU L,LIAO H,et al.Oxidative stress-induced differentiation of L6 myoblasts[J].Journal of First Military Medical University,2005,25(11):1384-1386.(in Chinese)
[3] FLANDIN P,DONATI Y,BARAZZONE-ARGIROFFO C,et al.Hyperoxia-mediated oxidative stress increases expression of UCP3 mRNA and protein in skeletal muscle[J].FEBS Lett,2005,579(16):3411-3415.
[4] SIMON H U,HAJ-YEHIA A,LEVI-SCHAFFER F.Role of reactive oxygen species (ROS) in apoptosis induction[J].Apoptosis, 2000,5(5):415-418.
[5] SCHERZ-SHOUVAL R,ELAZAR Z.Regulation of autophagy by ROS:physiology and pathology[J].Trends Biochem Sci,2011, 36(1):30-38.
[6] JIANG Y,ZHANG W H,GAO F,et al.Micro-encapsulated sodium butyrate attenuates oxidative stress induced by corticosterone exposure and modulates apoptosis in intestinal mucosa of broiler chickens[J].Anim Prod Sci,2014,55(5):587-594.
[7] YIN Y,LU L,WANG D T,et al.Astragalus polysaccharide inhibits autophagy and apoptosis from peroxide-induced injury in C2C12 myoblasts[J].Cell Biochem Biophys,2015,73(2):433-439.
[8] LI H,MALHOTRA S,KUMAR A.Nuclear factor-kappa B signaling in skeletal muscle atrophy[J].J Mol Med,2008, 86(10):1113-1126.
[9] 赵敏孟.胚蛋注射丙酮酸肌酸调控肉鸡能量代谢和肌肉发育的作用机理研究[D].南京:南京农业大学,2017.
ZHAO M M.Regulatory mechanism of in ovo feeding of creatine pyruvate on energy metabolism and muscle development of broiler chickens[D].Nanjing:Nanjing Agricultural University,2017.(in Chinese)
[10] LIVAK K J,SCHMITTGEN T D.Analysis of relative gene expression data using real-time quantitative PCR and the method[J].Methods,2001,25(4):402-408.
[11] 陈伟,林映才,马现永,等.一些抗氧化剂的抗/促氧化作用及其机制[J].动物营养学报,2012,24(4):595-605.
CHEN W,LIN Y C,MA X Y,et al.Anti/pro-oxidative functions of antioxidants and their mechanisms[J].Chinese Journal of Animal Nutrition,2012,24(4):595-605.(in Chinese)
[12] YIN J,REN W K,WU X S,et al.Oxidative stress-mediated signaling pathways:a review[J].J Food Agric Environ, 2013, 11(2):132-139.
[13] LEE J C,SON Y O,CHOI K C,et al.Hydrogen peroxide induces apoptosis of BJAB cells due to formation of hydroxyl radicals via intracellular iron-mediated Fenton chemistry in glucose oxidase-mediated oxidative stress[J].Mol Cells,2006,22(1):21-29.
[14] ESSICK E E,WILSON R M,PIMENTEL D R,et al.Adiponectin modulates oxidative stress-induced autophagy in cardiomyocytes[J]. PLoS One,2013,8(7):e68697.
[15] 韩飞,周孟良.过氧化氢诱导HepG2细胞产生氧化应激细胞模型的建立[J].食品科学,2011,32(5):55-57.
HAN F,ZHOU M L.An experimental HepG2 cell model of hydrogen peroxide induced DNA oxidative injury[J].Food Science, 2011,32(5):55-57.(in Chinese)
[16] YAZAKI T,HIRADATE Y,HOSHINO Y,et al.L-arnitine improves hydrogen peroxide-induced impairment of nuclear maturation in porcine oocytes[J].Anim Sci J,2013,84(5):395-402.
[17] 杨金莲.酒精刺激激活NF-κB信号通路促进肝癌的生长和转移[D].合肥:安徽医科大学,2012.
YANG J L.NF-κB activation mediates ethanol-promoted liver cancer progression[D].Hefei:Anhui Medical University,2012.(in Chinese)
[18] 程何祥,张荣庆,马颖艳,等.过氧化氢诱导乳鼠心肌细胞凋亡及半胱天冬酶-3抑制剂的作用[J].第四军医大学学报,2001,22(21):1953-1956.
CHENG H X,ZHANG R Q,MA Y Y,et al.Apoptosis of neonatal rat cardiomyocytes induced by exogenous hydroxyl free radical and the effect of caspase-3 antagonist[J].Journal of the Fourth Military Medical University,2001,22(21):1953-1956.(in Chinese)
[19] 肖卫民,蒋碧梅,石永忠,等.过氧化氢诱导C2C12细胞凋亡的机制初探[J].医学临床研究,2004,21(11):1288-1291.
XIAO W M,JIANG B M,SHI Y Z,et al.Preliminary investigation on the mechanism of C2C12 cell apoptosis induced by hydrogen peroxide[J].Journal of Clinical Research,2004,21(11):1288-1291.(in Chinese)
[20] JIANG B M,XIAO W M,SHI Y Z,et al.Role of Smac/DIABLO in hydrogen peroxide-induced apoptosis in C2C12 myogenic cells[J].Free Radical Bio Med,2005,39(5):658-667.
[21] WYLLIE A H."Where,O death,is thy sting?" A brief review of apoptosis biology[J].Mol Neurobiol,2010,42(1):4-9.
[22] KIFFIN R,BANDYOPADHYAY U,CUERVO A M. Oxidative stress and autophagy[J].Antioxid Redox Sign,2006,8(1-2):152-162.
[23] 易蓓,袁海汀,许永会,等.NF-κB抑制剂PDTC对人骨髓瘤U266细胞增殖和凋亡的影响[J].中国病理生理杂志, 2017,33(7):1177-1183.
YI B,YUAN H T,XU Y H,et al.Effect of NF-κB inhibitor pyrrolidine dithiocarbamate on proliferation and apoptosis of human multiple myeloma U266 cells[J].Chinese Journal of Pathophysiology,2017,33(7):1177-1183.(in Chinese)
[24] BYUN Y J,KIM S K,KIM Y M,et al.Hydrogen peroxide induces autophagic cell death in C6 glioma cells via BNIP3-mediated suppression of the mTOR pathway[J].Neurosci Lett,2009,461(2):131-135.
[25] ZHANG H Y,KONG X X,KANG J S,et al.Oxidative stress induces parallel autophagy and mitochondria dysfunction in human glioma U251 cells[J].Toxicol Sci,2009,110(2):376-388.
[26] YU L,WAN F Y,DUTTA S,et al.Autophagic programmed cell death by selective catalase degradation[J].Proc Natl Acad Sci USA,2006,103(13):4952-4957.
[27] SCHERZ-SHOUVAL R,SHVETS E,FASS E,et al.Reactive oxygen species are essential for autophagy and specifically regulate the activity of Atg4[J].EMBO J,2007,26(7):1749-1760.
[28] GAO H,LIN L,HAQ I U,et al.Inhibition of NF-κB promotes autophagy via JNK signaling pathway in porcine granulosa cells[J]. Biochem Biophys Res Commun,2016,473(1):311-316.
[29] YUAN L,WEI S P,WANG J,et al.Isoorientin induces apoptosis and autophagy simultaneously by reactive oxygen species (ROS)-related p53,PI3K/Akt,JNK,and p38 signaling pathways in HepG2 cancer cells[J].J Agric Food Chem,2014, 62(23):5390-5400.
[30] YIN J,DUAN J L,CUI Z J,et al.Hydrogen peroxide-induced oxidative stress activates NF-κB and Nrf2/Keap1 signals and triggers autophagy in piglets[J].RSC Adv,2015,5(20):15479-15486.
[31] 刘庆羊,孙自强,赵鸿雁,等.自噬在骨保护素对破骨细胞及其前体凋亡过程的调控机制[J].畜牧兽医学报,2018,49(8):1761-1769.
LIU Q Y,SUN Z Q,ZHAO H Y,et al.Effects of autophagy on the apoptosis of osteoclasts and osteoclast precursors mediated by osteoprotegerin[J].Acta Veterinaria et Zootechnica Sinica,2018,49(8):1761-1769.(in Chinese)
[32] 胡林,南良康,张敏,等.猪Beclin1基因调控伪狂犬病病毒诱导细胞自噬和凋亡的机制分析[J].畜牧兽医学报,2018,49(6):1274-1281.
HU L,NAN L K,ZHANG M,et al.The mechanism of porcine Beclin1 gene on autophagy and apoptosis[J].Acta Veterinaria et Zootechnica Sinica,2018,49(6):1274-1281.(in Chinese)
[33] 张勇,崔岩.NF-κB在细胞凋亡中的调节作用和应用前景[J].动物营养学报,2011,23(5):715-719.
ZHANG Y,CUI Y.NF-κB:modulation in apoptosis and application prospects[J].Chinese Journal of Animal Nutrition,2011, 23(5):715-719.(in Chinese)
[34] CHEN X X,ZHANG L,LI J L,et al.Hydrogen peroxide-induced change in meat quality of the breast muscle of broilers is mediated by ROS generation,apoptosis,and autophagy in the NF-κB signal pathway[J].J Agric Food Chem,2017, 65(19):3986-3994.
[35] CHEN X,GU R,ZHANG L,et al.Induction of nuclear factor-κB signal-mediated apoptosis and autophagy by reactive oxygen species is associated with hydrogen peroxide-impaired growth performance of broilers[J].Animal,2018,12(12):2561-2570.

Effects of H₂O₂ on Oxidative Damage of C2C12 Cells

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	LIU Jie, CONG Wei, ZHAO Mindie, ZHAO Ruqian. Expression of GR and FKBP5 in Hippocampus and Hypothalamus of Arbor Acres and Rugao Yellow Broiler Chickens and Its Relationship with Stress Sensitivity [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4766-4776.
[2]	DAI Lingli, LIU Zaixia, GUO Lili, YANG Yanda, CHANG Chencheng, WANG Yu, SHI Caixia, WANG Yuzhen, ZHANG Wenguang. β-Hydroxybutyrate Mediated Epigenetic Modification and Its Molecular Mechanism of Regulating Inflammation [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4095-4104.
[3]	ZHAO Huiying, YU Shiqiang, ZHAO Yuchao, JIANG Linshu. Mechanism of Liver-Adipose Tissue Crosstalk in the Development of Fatty Liver in Periparturient Cows [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4105-4116.
[4]	CAO Xiyue, JI Xiaoxia, ZHANG Chonghao, ZHANG Yafeng, CHEN Yutao, ZHANG Yuanshu. Diminazene Aceturate Alleviates Pulmonary Fibrosis in Mice by Endogenous Activation of ACE2 to Inhibit AngⅡ-TGF-β1 Pathway [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2631-2640.
[5]	YANG Heng, LI Licai, FU Lin, XU Huihao, ZHANG Dezhi, LI Qianyong. Effect of Prostaglandin D₂ on Endocrine Function and Expression of Apoptosis-related Genes in Goat Luteal Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1652-1663.
[6]	JIN Meilin, LI Taotao, SUN Dongxiao, WEI Caihong. Research Progress of Epigenetic Regulation in Fat Deposition Mechanism of Livestock and Poultry [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 855-867.
[7]	YE Qianwen, CHEN Zhuo, LI Xin, SUN Yawei, JIN Xiaoye, LI Ziqian, WUMAIERJIANG·Yahefu, ZHONG Qi, MA Xuelian, YAO Gang. Dynamic Changes of Gut Microbiota Composition and Its Predicted Metabolism Function in One-month-old Sucking Lambs [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1095-1108.
[8]	ZHANG Xiyu, ZHAI Pin, WANG Fan, DAI Yingying, ZHAO Bohao, CHEN Yang, WU Xinsheng. Expression and Function of KRT 16 in Hair Follicle Development of Angora Rabbit [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(1): 157-167.
[9]	LIU Zhihao, JIA Peng, LAI Qi, DONG Lifeng, WU Qiujue, GAO Yanhua, TIAN Zhonghong, DIAO Qiyu. Characteristics of Rumen Fermentation and Methane Production in Different Parity Dry Cows [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(12): 4296-4305.
[10]	CAO Xiyue, FU Yali, XIE Nana, ZHANG Yafeng, ZHANG Yuanshu. Inhibitory Effect and Mechanism of Endogenous Activation of Angiotensin Converting Enzyme 2 by Diminazene Aceturate on Cell Lipid Deposition [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2729-2738.
[11]	GUO Wenliang, XU Yuanqing, JIN Xiao, SHI Binlin. Moderating Role of Heat Shock Protein Under Inflammatory Response and Oxidative Stress Caused by Cold Stress [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(6): 1668-1677.
[12]	YUAN Jing, ZHANG Xinxin, ZHOU Di, YANG Shuai, ZHOU Jiayan, LIANG Aixin, YANG Liguo. Evaluation of the Effect of SS and CST Dual-expression DNA Vaccines with Attenuated Salmonella as Carrier in Immunizing Calves through Different Routes [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(6): 1914-1924.
[13]	SUN Yanjin, XUE Ya’nan, ZHONG Tao, WANG Linjie, LI Li, ZHANG Hongping, ZHAN Siyuan. The Function of HuR and Its Regulation on Muscle Growth and Development [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(5): 1345-1353.
[14]	DING Jun, FU Zilin, HE Junhao, DUAN Xiaowei, MA Lu, BU Dengpan. Research Progress of Milk-derived Exosomes [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(4): 1019-1029.
[15]	LI Yaqi, WANG Xianzhong, ZHANG Jiaojiao. Research Progress of Cell Proliferation and Apoptosis Regulated by Non-thermal Plasma and Its Application in Animal Husbandry [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(1): 1-10.

Effects of H2O2 on Oxidative Damage of C2C12 Cells

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

Effects of H₂O₂ on Oxidative Damage of C2C12 Cells