Email Alerts
RSS
Volume 39 Issue 5
May  2023
Turn off MathJax
Article Contents
Abstract
References
Article Navigation >  CHINESE JOURNAL OF BURNS AND WOUNDS  > 2023  >  39(5): 434-442
Lyu SJ,Yan ZD,Fan RH,et al.Effects and mechanism of glycine on rat cardiomyocytes pretreated with serum from burned rats[J].Chin J Burns Wounds,2023,39(5):434-442.DOI: 10.3760/cma.j.cn501225-20230206-00035.
Citation: Lyu SJ,Yan ZD,Fan RH,et al.Effects and mechanism of glycine on rat cardiomyocytes pretreated with serum from burned rats[J].Chin J Burns Wounds,2023,39(5):434-442.DOI: 10.3760/cma.j.cn501225-20230206-00035.
shu

Effects and mechanism of glycine on rat cardiomyocytes pretreated with serum from burned rats

doi: 10.3760/cma.j.cn501225-20230206-00035
  • 1.

    Department of Burns and Plastic Surgery, Shaanxi Provincial Armed Police Corps Hospital, Xi'an 710086, China

  • 2.

    Xi'an Beilin Tiansi Comprehensive Outpatient Department, Xi'an 710003, China

  • 3.

    Department of Burns and Plastic Surgery and Medical Cosmetic Surgery, Shaanxi Provincial People's Hospital, Xi'an 710068, China

  • 4.

    Clinical Medical Research Center, the First Affiliated Hospital of Army Medical University (the Third Military Medical University), Chongqing 400038, China

Funds:

General Program of National Natural Science Foundation of China 81971838

More Information
  • Corresponding author: Peng Xi, Email: pxlrmm@163.com
  • Received Date: 2023-02-06
    Abstract
  •   Objective   To investigate the effect and mechanism of glycine on rat cardiomyocytes pretreated with serum from burned rats (hereinafter referred to as burn serum).   Methods   Experimental research methods were adopted. Thirty gender equally balanced Wistar rats aged 7 to 8 weeks were collected, 10 of which were used to prepare normal rat serum (hereinafter referred to as normal serum), and the other 20 were inflicted with full-thickness burn of 30% total body surface area to prepare burn serum. Primary cardiomyocytes were isolated and cultured from the apical tissue of 180 Wistar rats aged 1 to 3 days by either gender for follow-up experiments. Cells were divided into normal serum group and burn serum group treated with corresponding serum according to the random number table (the same grouping method below). Trypanosoma blue staining was performed at post treatment hour (PTH) 1, 3, 6, 9, and 12 to detect the cell survival rate. Cells were divided into burn serum alone group treated with burn serum for 6 h followed by routine culture of 30 min and 0.4 mmol/L glycine group, 0.8 mmol/L glycine group, 1.2 mmol/L glycine group, 1.6 mmol/L glycine group, and 2.0 mmol/L glycine group treated with burn serum for 6 h followed by culture of 30 min with corresponding final molarity of glycine, i.e., at post intervention hour (PIH) 6.5, the cell survival rate was detected as before. Cells were divided into normal serum group, burn serum alone group, 0.8 mmol/L glycine group, 1.2 mmol/L glycine group, and 1.6 mmol/L glycine group, with the same intervention of 6.5 h as before, respectively. The content of adenosine monophosphate (AMP) and adenosine triphosphate (ATP) was detected by high performance liquid chromatography, and the AMP/ATP ratio was calculated. The protein expressions of phosphorylated mammalian target of rapamycin complex 1 (p-mTORC1), phosphorylated p70 ribosomal protein S6 kinase (p-p70 S6K), phosphorylated eukaryotic translation initiation factor 4E-binding protein 1 (p-4E-BP1), and phosphorylated AMP-activated protein kinase (p-AMPK) were detected by Western blotting. Cells were divided into normal serum group, burn serum alone group, 0.8 mmol/L glycine group intervened as before and 0.8 mmol/L glycine+25 ng/mL rapamycin group treated with burn serum followed by culture with two reagents. The expressions of heat shock protein 70 (HSP70), metallothionein (MT), and tubulin were detected by immunofluorescence method after 30 min of corresponding culture at PTH 1, 3, and 6, i.e., at PIH 1.5, 3.5, and 6.5, and the microtubule morphology was observed at PIH 6.5. The sample number at each time point was 10. Data were statistically analyzed with analysis of variance for factorial design, one-way analysis of variance, least significant difference (LSD)- t test, LSD test, and Bonferroni correction.   Results   At PTH 1, 3, 6, 9, and 12, the cell survival rates in burn serum group were significantly lower than those in normal serum group (with t values of 4.96, 16.83, 35.51, 34.33, and 27.88, P<0.05). In burn serum group, the cell survival rate at PTH 3, 6, 9, or 12 was significantly lower than that at PTH 1 ( P<0.05), the cell survival rate at PTH 6, 9, or 12 was significantly lower than that at PTH 3 ( P<0.05), and the cell survival rate at PTH 6 was similar to that at PTH 9 ( P>0.05) but significantly higher than that at PTH 12 ( P<0.05). Treatment of 6 h was selected as the follow-up intervention time of burn serum. At PIH 6.5, compared with that in burn serum alone group, the cell survival rate in each glycine group was significantly increased ( P<0.05). The cell survival rate in 0.8 mmol/L glycine group was the highest, and 0.8, 1.2, and 1.6 mmol/L were selected as subsequent glycine intervention concentrations. At PIH 6.5, the AMP/ATP ratio of cells in burn serum alone group was significantly higher than that in normal serum group, 1.2 mmol/L glycine group, or 1.6 mmol/L glycine group ( P values all <0.05), and the AMP/ATP ratio of cells in 1.6 mmol/L glycine group was significantly lower than that in 0.8 mmol/L glycine group ( P<0.05). At PIH 6.5, the protein expressions of p-mTORC1, p-p70 S6K, and p-4E-BP1 of cells in normal serum group, burn serum alone group, 0.8 mmol/L glycine group, 1.2 mmol/L glycine group, and 1.6 mmol/L glycine group were 1.001±0.037, 0.368±0.020, 1.153±0.019, 1.128±0.062, 1.028±0.037, 0.96±0.07, 0.63±0.12, 1.17±0.13, 1.13±0.16, 1.11±0.11, and 0.98±0.06, 0.45±0.08, 1.13±0.05, 0.77±0.12, 0.51±0.13. Compared with those in burn serum alone group, the protein expressions of p-mTORC1, p-p70 S6K, and p-4E-BP1 of cells in normal serum group and each glycine group were significantly increased ( P<0.05), while the protein expressions of p-AMPK were significantly decreased ( P<0.05). Compared with those in 0.8 mmol/L glycine group, the protein expression of p-4E-BP1 of cells in 1.2 mmol/L glycine group and the protein expressions of p-mTORC1 and p-4E-BP1 of cells in 1.6 mmol/L glycine group were significantly decreased ( P<0.05). Compared with those in 1.2 mmol/L glycine group, the protein expressions of p-mTORC1 and p-4E-BP1 of cells in 1.6 mmol/L glycine group were significantly decreased ( P<0.05), while the protein expression of p-AMPK was significantly increased ( P<0.05). Compared with those in normal serum group, the expression of tubulin of cells in burn serum alone group was significantly decreased at PIH 1.5, 3.5, and 6.5 ( P<0.05), while the expression of HSP70 of cells at PIH 1.5 and 3.5 and the expression of MT at PIH 3.5 and 6.5 were significantly increased ( P<0.05). The expressions of HSP70 and MT of cells at PIH 1.5, 3.5, and 6.5 and the expression of tubulin at PIH 1.5 and 3.5 in burn serum alone group and 0.8 mmol/L glycine+25 ng/mL rapamycin group were significantly lower than those in 0.8 mmol/L glycine group ( P<0.05). At PIH 6.5, compared with that in normal serum group, the cell microtubule structure in burn serum alone group was disordered; the cell boundary in 0.8 mmol/L glycine group was clearer than that in burn serum alone group, and the microtubule structure arranged neatly near the nucleus. Compared with that in 0.8 mmol/L glycine group, 0.8 mmol/L glycine+25 ng/mL rapamycin group had unclear cell boundaries and disordered microtubule structure.   Conclusions   Burn serum can cause cardiomyocytes damage in rats. Glycine can significantly up-regulate mammalian target of rapamycin/p70 ribosomal protein S6 kinase/eukaryotic translation initiation factor 4E-binding protein 1 signaling pathway through AMP-activated protein kinase, promote the synthesis of protective proteins HSP70, MT, and tubulin, stabilize the microtubule structure, and exert cardiomyocytes protection function.

     

    • FullText(HTML)
    • References(28)
  • [1]
    黄跃生.严重烧伤早期心肌损害机制及临床意义的再认识[J].中华烧伤杂志,2016,32(5):257-259.DOI: 10.3760/cma.j.issn.1009-2587.2016.05.001.
    [2]
    ClarkA,ImranJ,MadniT,et al.Nutrition and metabolism in burn patients[J/OL].Burns Trauma,2017,5:11[2023-02-06].https://pubmed.ncbi.nlm.nih.gov/28428966/.DOI: 10.1186/s41038-017-0076-x.
    [3]
    刘琰,王际壮.烧伤应激反应及其调控策略[J].中华烧伤杂志,2021,37(2):126-130.DOI: 10.3760/cma.j.cn501120-20201125-00499.
    [4]
    彭曦.烧伤临床营养新视角[J].中华烧伤杂志,2019,35(5):321-325.DOI: 10.3760/cma.j.issn.1009-2587.2019.05.001.
    [5]
    吕尚军,范荣辉,吴丹,等.谷氨酰胺对经烧伤大鼠血清干预的大鼠心肌细胞的作用及其细胞信号机制[J].中华烧伤杂志,2021,37(12):1149-1157.DOI: 10.3760/cma.j.cn501120-20210601-00208.
    [6]
    ZhangY,LvSJ,YanH,et al.Effects of glycine supplementation on myocardial damage and cardiac function after severe burn[J].Burns,2013,39(4):729-735.DOI: 10.1016/j.burns.2012.09.006.
    [7]
    MorioA,TsutsumiR,KondoT,et al.Leucine induces cardioprotection in vitro by promoting mitochondrial function via mTOR and Opa-1 signaling[J].Nutr Metab Cardiovasc Dis,2021,31(10):2979-2986.DOI: 10.1016/j.numecd.2021.06.025.
    [8]
    Ley-NgardigalS,BertolinG.Approaches to monitor ATP levels in living cells: where do we stand?[J].FEBS J,2022,289(24):7940-7969.DOI: 10.1111/febs.16169.
    [9]
    Costa-MattioliM,WalterP.The integrated stress response: from mechanism to disease[J].Science,2020,368(6489):eaat5314.DOI: 10.1126/science.aat5314.
    [10]
    OsborneT,WallB,EdgarDW,et al.Current understanding of the chronic stress response to burn injury from human studies[J/OL].Burns Trauma,2023,11:tkad007[2023-02-06].https://pubmed.ncbi.nlm.nih.gov/36926636/.DOI: 10.1093/burnst/tkad007.
    [11]
    SomeroGN.The cellular stress response and temperature: function, regulation, and evolution[J].J Exp Zool A Ecol Integr Physiol,2020,333(6):379-397.DOI: 10.1002/jez.2344.
    [12]
    LiuP,WangMZ,ZhongWX,et al.Stress-responsive genes (hsp70 and mt) and genotoxicity elicited by roxarsone exposure in Carassius auratus[J].Environ Toxicol Pharmacol,2018,62:132-139.DOI: 10.1016/j.etap.2018.07.004.
    [13]
    ZhongZ,WheelerMD,LiX,et al.L-glycine: a novel antiinflammatory, immunomodulatory, and cytoprotective agent[J].Curr Opin Clin Nutr Metab Care,2003,6(2):229-240.DOI: 10.1097/00075197-200303000-00013.
    [14]
    PanS,FanM,LiuZ,et al.Serine, glycine and one-carbon metabolism in cancer (Review)[J].Int J Oncol,2021,58(2):158-170.DOI: 10.3892/ijo.2020.5158.
    [15]
    GutP,LizzoG,MigliavaccaE,et al. Effects of glycine and n-acetylcysteine on glutathione levels and mitochondrial energy metabolism in healthy aging[J].Innov Aging,2021,5(Suppl 1):S685.DOI: 10.1093/geroni/igab046.2574.
    [16]
    SaxtonRA,SabatiniDM.mTOR signaling in growth, metabolism, and disease[J].Cell,2017,168(6):960-976.DOI: 10.1016/j.cell.2017.02.004.
    [17]
    ParkJH,LeeG,BlenisJ.Structural insights into the activation of mTORC1 on the lysosomal surface[J].Trends Biochem Sci,2020,45(5):367-369.DOI: 10.1016/j.tibs.2020.02.004.
    [18]
    Francois-VaughanH,AdebayoAO,BrilliantKE,et al.Persistent effect of mTOR inhibition on preneoplastic foci progression and gene expression in a rat model of hepatocellular carcinoma[J].Carcinogenesis,2016,37(4):408-419.DOI: 10.1093/carcin/bgw016.
    [19]
    LinSC,HardieDG.AMPK: sensing glucose as well as cellular energy status[J].Cell Metab,2018,27(2):299-313.DOI: 10.1016/j.cmet.2017.10.009.
    [20]
    SteinbergGR,CarlingD.AMP-activated protein kinase: the current landscape for drug development[J].Nat Rev Drug Discov,2019,18(7):527-551.DOI: 10.1038/s41573-019-0019-2.
    [21]
    ZhuJ,WangYF,ChaiXM,et al.Exogenous NADPH ameliorates myocardial ischemia-reperfusion injury in rats through activating AMPK/mTOR pathway[J].Acta Pharmacol Sin,2020,41(4):535-545.DOI: 10.1038/s41401-019-0301-1.
    [22]
    XiangF,MaSY,LvYL,et al.Tumor necrosis factor receptor-associated protein 1 regulates hypoxia-induced apoptosis through a mitochondria-dependent pathway mediated by cytochrome c oxidase subunit II[J/OL].Burns Trauma,2019,7:16[2023-02-06].https://pubmed.ncbi.nlm.nih.gov/31143823/.DOI: 10.1186/s41038-019-0154-3.
    [23]
    WangW,WuZ,DaiZ,et al.Glycine metabolism in animals and humans: implications for nutrition and health[J].Amino Acids,2013,45(3):463-477.DOI: 10.1007/s00726-013-1493-1.
    [24]
    BuruteM,KapiteinLC.Cellular logistics: unraveling the interplay between microtubule organization and intracellular transport[J].Annu Rev Cell Dev Biol,2019,35:29-54.DOI: 10.1146/annurev-cellbio-100818-125149.
    [25]
    YuanAT,KorkolaNC,WongDL,et al.Metallothionein Cd4S11 cluster formation dominates in the protection of carbonic anhydrase[J].Metallomics,2020,12(5):767-783.DOI: 10.1039/d0mt00023j.
    [26]
    谢果,陈伟,王海英.金属硫蛋白在心肌缺血/再灌注损伤中保护作用的研究进展[J].国际麻醉学与复苏杂志,2021,42(9):1004-1008.DOI: 10.3760/cma.j.cn321761-20200601-00382.
    [27]
    YangL,MaJ,TanY,et al.Cardiac-specific overexpression of metallothionein attenuates L-NAME-induced myocardial contractile anomalies and apoptosis[J].J Cell Mol Med,2019,23(7):4640-4652.DOI: 10.1111/jcmm.14375.
    [28]
    KuznetsovAV,JavadovS,GrimmM,et al.Crosstalk between mitochondria and cytoskeleton in cardiac cells[J].Cells,2020,9(1):222.DOI: 10.3390/cells9010222.
    • Relative Articles

  • Relative Articles

    [1]Zou Wen, Han Chunmao, Jin Ronghua, Shen Tao. Selection and analysis of calculation formulas for resting energy expenditure in patients with severe burns based on different metabolic stages[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2024, 40(7): 634-642. doi: 10.3760/cma.j.cn501225-20240229-00080
    [2]Lyu Shangjun, Fan Ronghui, Wu Dan, Peng Xi. Effects and cell signaling mechanism of glutamine on rat cardiomyocytes intervened with serum from burned rat[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2021, 37(12): 1149-1157. doi: 10.3760/cma.j.cn501120-20210601-00208
    [3]Xiang Fei, Xue Dongdong, Luo Jia, Hu Jianhong, Yuan Lili, Jia Jiezhi, Huang Yuesheng. Effects and mechanism of mitochondrial transcription factor A and cytochrome c oxidase pathway in the energy production of hypoxic cardiomyocytes of rats regulated by tumor necrosis factor receptor associated protein 1[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2020, 36(8): 651-657. doi: 10.3760/cma.j.cn501120-20200430-00247
    [4]Yang Lei, Zhao Liping, Cui Lin, Huang Yao, Ye Jingying, Zhang Qiong, Zhang Dongxia, Huang Yuesheng. Effects of decline of pH value on cardiomyocyte viability of rats and the mechanism[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2018, 34(5): 303-308. doi: 10.3760/cma.j.issn.1009-2587.2018.05.011
    [5]Xiang Fei, Zhang Dongxia, Ma Siyuan, Huang Yuesheng. Mechanism of protective effects of tumor necrosis factor receptor associated protein 1 on hypoxic cardiomyocytes of rats[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2016, 32(12): 744-751. doi: 10.3760/cma.j.issn.1009-2587.2016.12.010
    [6]Lan Xiaodong, Dang Yongming, Li Lingfei, Zhang Qiong, Huang Yuesheng. Effects of microtubule depolymerization on spontaneous beating and action potential of cardiac myocytes in rats and its mechanism[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2015, 31(3): 192-198. doi: 10.3760/cma.j.issn.1009-2587.2015.03.009
    [7]ZHENG Jun, HUANG Yue-sheng, HUANG Xiao-yuan, FAN Peng-ju, HE Wei-feng, ZHANG Xiao-rong. Effects of antisense p38a mitogen-activated protein kinase on myocardial cells exposed to hypoxia and burn serum[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2013, 29(3): 267-271. doi: 10.3760/cma.j.issn.1009-2587.2013.03.010
    [8]DANG Yong-ming, FANG Ya-dong, HU Jiong-yu, ZHANG Jia-ping, SONG Hua-pei, ZHANG Yi-ming, ZHANG Qiong, HUANG Yue-sheng. Influence of microtubule depolymerization of myocardial cells on mitochondria distribution and ener- gy metabolism in adult rats[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2010, 26(1): 18-22. doi: 10.3760/cma.j.issn.1009-2587.2010.01.006
    [9]TENG Miao, HUANG Yue-sheng, DANG Yong-ming, FANG Ya-dong, ZHANG Qiong. The influence of microtubule intervention drugs on glycolytic key enzymes in myocardial cells after hypoxia[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2008, 24(2): 102-106.
    [10]YUAN Zhi-qiang, LI Xiao-lu, PENG Yi-zhi, WANG Pei, HUANG Yue-sheng, YANG Zong-cheng. Influence of HSP70 on function and energy metabolism of mitochondria in intestinal epithelial cells after hypoxia/reoxygenation[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2008, 24(3): 203-206.
    [11]ZHOU Jun-li, HUANG Yue-sheng, SONG Hua-pei, DANG Yong-ming, ZHANG Dong-xia, ZHANG Qiong. Effects of Glyeine on apoptosis in murine cardiomyocyte suffering from ischemia and hypoxia[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2008, 24(3): 167-170.
    [12]HU Jiong-yu, HUANG Yue-sheng, SONG Hua-pei, ZHANG Dong-xia, XIANG Fei, ZHU Zhi-gang, TENG Miao, ZHANG Qiong. Protective effects of Astragaloside and Quercetin on rat myocardial cells after hypoxia[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2007, 23(3): 175-178.
    [13]KUANG Yong, HUANG Yue-sheng. Effect of the microtubule depolymerization on mitochondria damage in rat myocadiocytes early after hypoxia[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2007, 23(4): 288-291.
    [14]KUANG Yong, HUANG Yue-sheng. Study on injury to microtubule of cardiomyosites at early post-hypoxia stage[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2007, 23(3): 172-174.
    [15]LV Shang-jun, ZHANG Yong, SUN Yong, WU Wei, YOU Zhong-yi, WANG Shi-liang, PENG Xi. Protective effects of glycyl-glutamine dipeptide supplement on the heart function in burn rats[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2007, 23(4): 244-248.
    [16]TENG Miao, HUANG Yue-sheng, ZHENG Ji, DANG Yong-ming, ZHANG Qiong. The influence of microtubule intervention drugs on the energy metabolism of myocardial cells after hypoxia[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2007, 23(3): 164-167.
    [17]ZHENG Ji, FANG Ya-dong, TENG Miao, DANG Yong-ming, KUANG Yong, YAN Hong, ZHANG Dong-xia, SONG Hua-pei, ZHANG Qiong, HUANG Yue-sheng. Study on the influence of hypoxia induced microtubule damage on the opening of mitochondriai permeable transition pore of cardiac myocytes in rat[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2006, 22(3): 195-198.
    [18]ZHOU Jun-li, HUANG Yue-sheng, DANG Yong-ming, ZHANG Jia-ping. Protective effect of glycin on hypoxic rat myocardial cells[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2005, 21(5): 329-332.
    [19]GAO Zhi-gang, LIU Qun, XIE Yu-gang, WANG Yu-lian, DENG Shi-lin. Modulation Effects of Recombinant Human Growth Hormone on Postburn Hypermetabolism in Burn Patients[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2004, 20(3): 158-160.
    [20]YAO Qing-jun, JIA Chi-yu, CHEN Bi, LU Ning, XU Ming-da, LIU Xin-ping, YAO LI-bo, WANG Shu-sen. Expression of microtubule destabilizer-oncoprotein18/stathmin gene in the hypertrophic scar before and after the healing of the deep partial thickness burn wounds[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2003, 19(Z1): 15-17. doi: 10.3760/cma.j.issn.1009-2587.2003.Z1.107
    • Supplements(0)
    • Cited By
  • 加载中

Catalog

    通讯作者: 陈斌, bchen63@163.com
    • 1. 

      沈阳化工大学材料科学与工程学院 沈阳 110142

    1. 本站搜索
    2. 百度学术搜索
    3. 万方数据库搜索
    4. CNKI搜索

    Figures(3)  / Tables(3)

    Get Citation
    PDF
    XML

    Article Metrics

    Article has an altmetric score of 1
    Article views (2157) PDF downloads(11) Cited by()
    Proportional views
    Related

    Copyright © Chinese Journal of Burns京ICP备07035254号-14

    E-mail：shaoshangzazhi@163.com

    Website：https://zhsszz.xml-journal.net/

    Supported by: Beijing Renhe Information Technology Co. Ltd    

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return