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Volume 38 Issue 7
Jul.  2022
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Article Navigation >  CHINESE JOURNAL OF BURNS AND WOUNDS  > 2022  >  38(7): 667-676
Fan FX,Li PT,Xia ZG,et al.Effects and molecular mechanism of exogenous L-carnitine on excessive endoplasmic reticulum stress-mediated hepatic pyroptosis in severely scald rats[J].Chin J Burns Wounds,2022,38(7):667-676.DOI: 10.3760/cma.j.cn501225-20220120-00010.
Citation: Fan FX,Li PT,Xia ZG,et al.Effects and molecular mechanism of exogenous L-carnitine on excessive endoplasmic reticulum stress-mediated hepatic pyroptosis in severely scald rats[J].Chin J Burns Wounds,2022,38(7):667-676.DOI: 10.3760/cma.j.cn501225-20220120-00010.
shu

Effects and molecular mechanism of exogenous L-carnitine on excessive endoplasmic reticulum stress-mediated hepatic pyroptosis in severely scald rats

doi: 10.3760/cma.j.cn501225-20220120-00010
  • 1.

    Department of Burn Surgery, the First Affiliated Hospital of Anhui Medical University, Hefei 230022, China

  • 2.

    Department of Plastic Surgery, the First Affiliated Hospital of Anhui Medical University, Hefei 230022, China

  • 3.

    Department of Burns and Plastic Surgery, the Fourth Affiliated Hospital of Anhui Medical University, Hefei 230022, China

  • 4.

    Department of Immunology, School of Basic Medicine, Anhui Medical University, Hefei 230032, China

Funds:

Key Science and Technology Project in Anhui Province of China 1604a0802083

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    Abstract
  •   Objective  To investigate the effects and molecular mechanism of exogenous L-carnitine on hepatic pyroptosis mediated by excessive endoplasmic reticulum stress in severely scald rats.  Methods  The experimental research method was adopted. According to the random number table (the same group method below), fifteen female Sprague Dawley rats aged 6-8 weeks were divided into sham-injury group, scald alone group, and scald+carnitine group (with 5 rats in each group), and full-thickness scald of 30% total body surface area were made on the back of rats in scald alone group and scald+carnitine group, and rats in scald+carnitine group were additionally given intraperitoneal injection of L-carnitine. At post injury hour (PIH) 72, The levels of aspartate aminotransferase (AST) and alanine dehydrogenase (ALT) of biochemical indicators of liver injury were detected by automatic biochemical analyzer with the sample number of 5. At PIH 72, liver tissue damage was detected by hematoxylin-eosin staining. At PIH 72, The mRNA levels of nucleotide-binding oligomerization domain-containing protein-like receptor family pyrin domain containing 3 (NLRP3), cysteine aspartic acid specific protease 1 (caspase-1), gasderminD (GSDMD), and interleukin 1β(IL-1β) in liver tissue as pyroptosis-related markers and glucose regulatory protein 78 (GRP78) and CCAAT/enhancer binding protein homologous protein (CHOP) in liver tissue as endoplasmic reticulum stress-related markers were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-qPCR). Protein expression levels of GRP78, CHOP, NLRP3, caspase-1, caspase-1/p20, GSDMD-N, and cleaved IL-1β in liver tissue were detected by Western blotting, and the sample numbers were all 5. HepG2 cells as human liver cancer cells were divided into dimethyl sulfoxide (DMSO) group, 0.1 μmol/L tunicamycin (TM) group, 0.2 μmol/L TM group, 0.4 μmol/L TM group, and 0.8 μmol/L TM group and were treated accordingly. After 24 h of culture, cell viability was detected by cell counting kit 8, and the intervention concentration of TM was screened, and the sample number was 5. HepG2 cells were divided into DMSO group, TM alone group, and TM+carnitine group, and treated accordingly. After 24 h of culture, the protein expression levels of GRP78, CHOP, NLRP3, caspase-1, caspase-1/p20, GSDMD-N, and cleaved IL-1β in cells were detected by Western blotting, and the sample numbers were all 3. Data were statistically analyzed with one-way analysis of variance and least significant difference-t test.  Results  At PIH 72, the AST and ALT levels of serum in scald alone group were (640±22) and (157±8) U/L, which were significantly higher than (106±13) and (42±6) U/L in sham-injury group, respectively, with t values of -46.78 and -25.98, respectively, P<0.01. The AST and ALT levels of serum in scald+carnitine group were (519±50) and (121±10) U/L, which were significantly lower than those in scald alone group, respectively, with t values of 4.93 and 6.06, respectively, P<0.01. At PIH 72, the morphology of liver tissue of rats in sham-injury group were basically normal with no obvious inflammatory cell infiltration; compared with those in sham-injury group, the liver tissue of rats in scald alone group showed a large number of inflammatory cell infiltration and disturbed cell arrangement; compared with that in scald alone group, the liver tissue of rats in scald+carnitine group showed a small amount of inflammatory cell infiltration. At PIH 72, the mRNA expression on levels of NLRP3, caspase-1, GSDMD, and IL-1β in liver tissue of rats in scald alone group were significantly higher than those in sham-injury group (with t values of 34.42, 41.93, 30.17, and 15.68, respectively, P<0.01); the mRNA levels of NLRP3, caspase-1, GSDMD, and IL-1β in liver tissue of rats in scald+carnitine group were significantly lower than those in scald alone group (with t values of 34.40, 37.20, 19.95, and 7.88, respectively, P<0.01). At PIH 72, the protein expression levels of NLRP3, caspase-1, caspase-1/p20, GSDMD-N, and cleaved IL-1β in liver tissue of rats in scald alone group were significantly higher than those in sham-injury group (with t values of 12.28, 26.92, 5.20, 10.02, and 24.78, respectively, P<0.01); compared with those in scald alone group, the protein expression levels of NLRP3, caspase-1, caspase-1/p20, GSDMD-N, and cleaved IL-1β in liver tissue of rats in scald+carnitine group were significantly decreased (with t values of 10.99, 27.96, 12.69, 8.96, and 12.27, respectively, P<0.01). At PIH 72, the mRNA levels of GRP78 and CHOP in liver tissue of rats in scald alone group were significantly higher than those in sham-injury group (with t values of 21.00 and 16.52, respectively, P<0.01), and the mRNA levels of GRP78 and CHOP in liver tissue of rats in scald+carnitine group were significantly lower than those in scald alone group (with t values of 8.92 and 8.21, respectively, P<0.01); the protein expression levels of GRP78 and CHOP in liver tissue of rats in scald alone group were significantly higher than those in sham-injury group (with t values of 22.50 and 14.29, respectively, P<0.01), and the protein expression levels of GRP78 and CHOP in liver tissue of rats in scald+carnitine group were significantly lower than those in scald alone group (with t values of 14.29 and 5.33 respectively, P<0.01). After 24 h of culture, the cell survival rates of 0.1 μmol/L TM group, 0.2 μmol/L TM group, 0.4 μmol/L TM group, and 0.8 μmol/L TM group were significantly decreased than that in DMSO group (with t values of 4.90, 9.35, 18.64, and 25.09, respectively, P<0.01). Then 0.8 μmol/L was selected as the intervention concentration of TM. After 24 h of culture, compared with that in DMSO group, the protein expression levels of GRP78 and CHOP in cells in TM alone group were significantly increased (with t values of 10.48 and 17.67, respectively, P<0.01), and the protein expression levels of GRP78 and CHOP in TM+carnitine group were significantly lower than those in TM alone group (with t values of 8.08 and 13.23, respectively, P<0.05 or P<0.01). After 24 h of culture, compared with those in DMSO group, the protein expression levels of NLRP3 and GSDMD-N in cells in TM alone group were significantly increased (with t values of 13.44 and 27.51, respectively, P<0.01), but the protein expression levels of caspase-1, caspase-1/p20, and cleaved IL-1β in cells were not significantly changed (P>0.05); compared with that in TM alone group, the protein expression levels of NLRP3 and GSDMD-N in cells in TM+carnitine group were significantly decreased (with t values of 20.49 and 21.95, respectively, P<0.01), but the protein expression levels of caspase-1, caspase-1/p20, and cleaved IL-1β in cells were not significantly changed (P>0.05).  Conclusions  In severely scald rats, exogenous L-carnitine may play a protective role against liver injury by inhibiting the pathways related to excessive endoplasmic reticulum stress-mediated pyroptosis.

     

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    • References(34)
  • [1]
    PorterC, TompkinsRG, FinnertyCC, et al. The metabolic stress response to burn trauma: current understanding and therapies[J]. Lancet, 2016,388(10052):1417-1426. DOI: 10.1016/S0140-6736(16)31469-6.
    [2]
    JeschkeMG, MicakRP, FinnertyCC, et al. Changes in liver function and size after a severe thermal injury[J]. Shock, 2007,28(2):172-177. DOI: 10.1097/shk.0b013e318047b9e2.
    [3]
    PriceLA, ThombsB, ChenCL, et al. Liver disease in burn injury: evidence from a national sample of 31,338 adult patients[J]. J Burns Wounds, 2007,7:e1.
    [4]
    褚万立, 柴家科, 冯永强, 等. 严重烧伤大鼠肝脏内质网应激相关蛋白的表达及其意义[J].中华医学杂志,2012,92(12):853-856. DOI: 10.3760/cma.j.issn.0376-2491.2012.12.019.
    [5]
    StanojcicM, VinaikR, AbdullahiA, et al. NLRP3 knockout enhances immune infiltration and inflammatory responses and improves survival in a burn sepsis model[J]. Immunology, 2022,165(2):195-205. DOI: 10.1111/imm.13427.
    [6]
    Al MamunA, WuY, JiaC, et al. Role of pyroptosis in liver diseases[J]. Int Immunopharmacol, 2020,84:106489. DOI: 10.1016/j.intimp.2020.106489.
    [7]
    SadigharaM, JoktajiJP, HajhashemiV, et al. Protective effects of coenzyme Q(10) and L-carnitine against statin-induced pancreatic mitochondrial toxicity in rats[J]. Res Pharm Sci, 2017,12(6):434-443. DOI: 10.4103/1735-5362.217424.
    [8]
    BusquetsS, SerpeR, ToledoM, et al. L-carnitine: an adequate supplement for a multi-targeted anti-wasting therapy in cancer[J]. Clin Nutr, 2012,31(6):889-895. DOI: 10.1016/j.clnu.2012.03.005.
    [9]
    HotaKB, HotaSK, ChaurasiaOP, et al. Acetyl-L-carnitine- mediated neuroprotection during hypoxia is attributed to ERK1/2-Nrf2-regulated mitochondrial biosynthesis[J]. Hippocampus, 2012,22(4):723-736. DOI: 10.1002/hipo.20934.
    [10]
    PatelSP, SullivanPG, LyttleTS, et al. Acetyl-L-carnitine ameliorates mitochondrial dysfunction following contusion spinal cord injury[J]. J Neurochem, 2010,114(1):291-301. DOI: 10.1111/j.1471-4159.2010.06764.x.
    [11]
    XiaZ, TianX, KongW, et al. Effects of L-carnitine on liver injury in rats and its impact on blood lipids[J]. Int J Clin Exp Med, 2018, 11(9):9768-9773.
    [12]
    LiP, XiaZ, KongW, et al. Exogenous L-carnitine ameliorates burn-induced cellular and mitochondrial injury of hepatocytes by restoring CPT1 activity[J]. Nutr Metab (Lond), 2021,18(1):65. DOI: 10.1186/s12986-021-00592-x.
    [13]
    徐晓东, 邓洋洋, 郑洪新. 糖皮质激素诱导肾虚骨质疏松症大鼠骨组织中OPG/RANKLmRAN及蛋白表达的影响[J].中华中医药学刊,2013,31(12):2623-2627.
    [14]
    中华医学会骨质疏松和骨矿盐疾病分会. 骨代谢生化标志物临床应用指南[J].中华骨质疏松和骨矿盐疾病杂志,2015,8(4):283-293. DOI: 10.3969/j.issn.1674-2591.2015.04.001.
    [15]
    苏延林, 宁斌. 骨质疏松的骨转换生物标志物及其临床应用[J/CD].转化医学电子杂志,2017,4(6):64-68. DOI: 10.3969/j.issn.2095-6894.2017.06.016.
    [16]
    NagyV, PenningerJM. The RANKL-RANK story[J]. Gerontology, 2015,61(6):534-542. DOI: 10.1159/000371845.
    [17]
    JeschkeMG, FinnertyCC, HerndonDN, et al. Severe injury is associated with insulin resistance, endoplasmic reticulum stress response, and unfolded protein response[J]. Ann Surg, 2012,255(2):370-378. DOI: 10.1097/SLA.0b013e31823e76e7.
    [18]
    RochaJ, Eduardo-FigueiraM, BarateiroA, et al. Erythropoietin reduces acute lung injury and multiple organ failure/dysfunction associated to a scald-burn inflammatory injury in the rat[J]. Inflammation, 2015,38(1):312-326. DOI: 10.1007/s10753-014-0035-7.
    [19]
    YangX, BaiH, CaiW, et al. Inhibition of Na+/H+ exchanger 1 by cariporide alleviates burn-induced multiple organ injury[J]. J Surg Res, 2013,185(2):797-804. DOI: 10.1016/j.jss.2013.06.049.
    [20]
    孔维昌, 夏正国, 田曦茜, 等. 早期补充外源性肉碱对严重烫伤大鼠肝脏线粒体损伤的影响及病理机制[J].中华烧伤杂志,2018,34(6):374-379. DOI: 10.3760/cma.j.issn.1009-2587.2018.06.012.
    [21]
    BortolottiP, FaureE, KipnisE. Inflammasomes in tissue damages and immune disorders after trauma[J]. Front Immunol, 2018,9:1900. DOI: 10.3389/fimmu.2018.01900.
    [22]
    WangZ, ZhangS, XiaoY, et al. NLRP3 inflammasome and inflammatory diseases[J]. Oxid Med Cell Longev, 2020,2020:4063562. DOI: 10.1155/2020/4063562.
    [23]
    Fernandes-AlnemriT, WuJ, YuJW, et al. The pyroptosome: a supramolecular assembly of ASC dimers mediating inflammatory cell death via caspase-1 activation[J]. Cell Death Differ, 2007,14(9):1590-1604. DOI: 10.1038/sj.cdd.4402194.
    [24]
    ShiJ, ZhaoY, WangK, et al. Cleavage of GSDMD by inflammatory caspases determines pyroptotic cell death[J]. Nature, 2015,526(7575):660-665. DOI: 10.1038/nature15514.
    [25]
    D'SouzaCA, HeitmanJ. Dismantling the cryptococcus coat[J]. Trends Microbiol, 2001,9(3):112-113.DOI: 10.1016/s0966-842x(00)01945-4.
    [26]
    HagarJA, PowellDA, AachouiY, et al. Cytoplasmic LPS activates caspase-11: implications in TLR4-independent endotoxic shock[J]. Science, 2013,341(6151):1250-1253. DOI: 10.1126/science.1240988.
    [27]
    KayagakiN, WongMT, StoweIB, et al. Noncanonical inflammasome activation by intracellular LPS independent of TLR4[J]. Science, 2013,341(6151):1246-1249. DOI: 10.1126/science.1240248.
    [28]
    ShiJ, ZhaoY, WangY, et al. Inflammatory caspases are innate immune receptors for intracellular LPS[J]. Nature, 2014,514(7521):187-192. DOI: 10.1038/nature13683.
    [29]
    SudsawardS, KhunchaiS, ThepmaleeC, et al. Endoplasmic reticulum stress, unfolded protein response and autophagy contribute to resistance to glucocorticoid treatment in human acute lymphoblastic leukaemia cells[J]. Int J Oncol, 2020,57(3):835-844. DOI: 10.3892/ijo.2020.5089.
    [30]
    NybergWA, EspinosaA. Imiquimod induces ER stress and Ca2+ influx independently of TLR7 and TLR8[J]. Biochem Biophys Res Commun, 2016,473(4):789-794. DOI: 10.1016/j.bbrc.2016.03.080.
    [31]
    JägerR, BertrandMJ, GormanAM, et al. The unfolded protein response at the crossroads of cellular life and death during endoplasmic reticulum stress[J]. Biol Cell, 2012,104(5):259-270. DOI: 10.1111/boc.201100055.
    [32]
    LeeAS. The ER chaperone and signaling regulator GRP78/BiP as a monitor of endoplasmic reticulum stress[J]. Methods, 2005,35(4):373-381. DOI: 10.1016/j.ymeth.2004.10.010.
    [33]
    QiZ, ChenL. Endoplasmic reticulum stress and autophagy[J]. Adv Exp Med Biol, 2019, 1206:167-177. DOI: 10.1007/978-981-15-0602-4_8.
    [34]
    NovoaI, ZengH, HardingHP, et al. Feedback inhibition of the unfolded protein response by GADD34-mediated dephosphorylation of eIF2alpha[J]. J Cell Biol, 2001, 153: 1011-1022.DOI: 10.1083/jcb.153.5.1011.
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