积雪草酸通过抑制氧化应激和铁死亡减轻脂多糖诱导的肉鸡心肌损伤的研究

doi:10.11843/j.issn.0366-6964.2024.04.040

Abstract

Abstract: This experiment was conducted to investigate the effects of different concentrations of asiatic acid (AA)on lipopolysaccharide (LPS)-induced acute myocardial injury (AMI) in broilers. All 50 broilers were selected to be reared to 7 days of age, and then divided into 5 groups, namely Control group, LPS group, LPS+AA 15 mg·kg^-1 group, LPS+AA 30 mg·kg^-1 group, LPS+AA 60 mg·kg^-1 group. At 7 days of age, all the AA groups were pretreated with corresponding doses of AA by gavage for 14 consecutive days. At 16, 18 and 20 days of age, all the LPS groups were intraperitoneally injected with 0.5 mg·kg^-1 LPS to induce AMI. Myocardial samples of broilers were collected at the age of 21 days. Histopathological changes of myocardium were observed by hematoxylin-eosin(HE)staining; Detection of markers of oxidative stress in myocardial tissue using glutathione peroxidase (GSH-Px)and malondialdehyde (MDA)kits; Real-time fluorescence PCR(qRT-PCR)and Western blot were used to detect the expression of genes and proteins related to oxidative stress and ferroptosis in myocardial tissues. The positive expression rate of Nrf2 and SLC7A11 in myocardial tissues was detected by immunohistochemistry. The positive expression rate of GPX4 in myocardial tissues was detected by immunofluorescence. The results showed that AA pretreatment alleviated the myocarial injury induced by LPS. Compared with LPS group, medium-dose AA pretreatment increased GSH-Px activity while all AA pretreatment decreased MDA content in myocardium significantly (P < 0.05 or P < 0.01). Importantly, AA pretreatment decreased the mRNA expression of Keap1, PTGS2 and HMGB1, and increased the expression of Nrf2, HO-1, NQO1, FTH1, SLC7A11, GPX4, GCLC and GCLM (P < 0.05 or P < 0.01). AA pretreatment also decreased the protein expression of Keap1 and HMGB1, promoted the protein expression of Nrf2, HO-1, NQO1, FTH1 and GPX significantly (P < 0.05). Immunohistochemistry and immunofluorescence results showed that AA pretreatment could significantly increased the expression of Nrf2 (P < 0.01), SLC7A11 (P < 0.01) and GPX (P < 0.01). The results of the current study showed for the first time that AA could alleviate LPS-induced AMI by regulating oxidative stress and ferroptosis. AA is expected to be a potential food additive for preventing LPS-induced AMI in broilers in the future.

Key words: asiatic acid, acute myocardial injury, lipopolysaccharide, oxidative stress, ferroptosis

CLC Number:

S858.31

ZHANG Xinting, QIU Wenyue, PANG Xiaoyue, SU Yiman, YE Jiali, HUANG Jianjia, ZHOU Shuilian, TANG Zhaoxin, WANG Rongmei, SU Rongsheng. Effect of Asiatic Acid Alleviating Myocardial Injury Caused by Lipopolysaccharide through Inhibiting Oxidative Stress and Ferroptosis in Broilers[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1787-1799.

References

[1] HOTCHKISS R S, MONNERET G, PAYEN D. Sepsis-induced immunosuppression:from cellular dysfunctions to immunotherapy[J]. Nat Rev Immunol, 2013, 13(12):862-874.
[2] LANDESBERG G, GILON D, MEROZ Y, et al. Diastolic dysfunction and mortality in severe sepsis and septic shock[J]. Eur Heart J, 2012, 33(7):895-903.
[3] DOS SANTOS J L, DE QUADROS A S, WESCHENFELDER C, et al. Oxidative stress biomarkers, nut-related antioxidants, and cardiovascular disease[J]. Nutrients, 2020, 12(3):682.
[4] BARRERA G, PIZZIMENTI S, DAGA M, et al. Lipid peroxidation-derived aldehydes, 4-hydroxynonenal and malondialdehyde in aging-related disorders[J]. Antioxidants (Basel), 2018, 7(8):102.
[5] BRIGELIUS-FLOHÉ R, MAIORINO M. Glutathione peroxidases[J]. Biochim Biophys Acta (BBA)-Gen Subj, 2013, 1830(5):3289-3303.
[6] SUN X F, OU Z H, CHEN R C, et al. Activation of the p62-Keap1-NRF2 pathway protects against ferroptosis in hepatocellular carcinoma cells[J]. Hepatology, 2016, 63(1):173-184.
[7] BELLEZZA I, GIAMBANCO I, MINELLI A, et al. Nrf2-Keap1 signaling in oxidative and reductive stress[J]. Biochim Biophys Acta (BBA)-Mol Cell Res, 2018, 1865(5):721-733.
[8] STOCKWELL B R, FRIEDMANN ANGELI J P, BAYIR H, et al. Ferroptosis:a regulated cell death nexus linking metabolism, redox biology, and disease[J]. Cell, 2017, 171(2):273-285.
[9] QIAO H Y, SAI X Y, GAI L Y, et al. Association between heme oxygenase 1 gene promoter polymorphisms and susceptibility to coronary artery disease:a huge review and meta-analysis[J]. Am J Epidemiol, 2014, 179(9):1039-1048.
[10] LI X G, LIU Z D, ZHANG A L, et al. NQO1 targeting prodrug triggers innate sensing to overcome checkpoint blockade resistance[J]. Nat Commun, 2019, 10(1):3251.
[11] CHEN Y, DONG H, THOMPSON D C, et al. Glutathione defense mechanism in liver injury:insights from animal models[J]. Food Chem Toxicol, 2013, 60:38-44.
[12] WANG K X, GAO S Y, WANG J R, et al. Protective effects of chicoric acid on LPS-induced endometritis in mice via inhibiting ferroptosis by Nrf2/HO-1 signal axis[J]. Int Immunopharmacol, 2022, 113:109435.
[13] MAO P, WANG Z H, LI J J, et al. Research progress of ferroptosis in bacterial infection[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6):2280-2287. (in Chinese)
毛鹏, 王志浩, 李建基, 等. 铁死亡在细菌性感染中的研究进展[J]. 畜牧兽医学报, 2023, 54(6):2280-2287.
[14] DIXON S J, LEMBERG K M, LAMPRECHT M R, et al. Ferroptosis:an iron-dependent form of nonapoptotic cell death[J]. Cell, 2012, 149(5):1060-1072.
[15] ZHANG Q Z, WU H M, ZOU M Y, et al. Folic acid improves abnormal behavior via mitigation of oxidative stress, inflammation, and ferroptosis in the BTBR T+ tf/J mouse model of autism[J]. J Nutr Biochem, 2019, 71:98-109.
[16] DOLL S, PRONETH B, TYURINA Y Y, et al. ACSL4 dictates ferroptosis sensitivity by shaping cellular lipid composition[J]. Nat Chem Biol, 2017, 13(1):91-98.
[17] XU C X, SUN S G, JOHNSON T, et al. The glutathione peroxidase Gpx4 prevents lipid peroxidation and ferroptosis to sustain Treg cell activation and suppression of antitumor immunity[J]. Cell Rep, 2021, 35(11):109235.
[18] FANG X X, CAI Z X, WANG H, et al. Loss of cardiac ferritin H facilitates cardiomyopathy via slc7a11-mediated ferroptosis[J]. Circ Res, 2020, 127(4):486-501.
[19] DU J, WANG T T, LI Y C, et al. DHA inhibits proliferation and induces ferroptosis of leukemia cells through autophagy dependent degradation of ferritin[J]. Free Radic Biol Med, 2019, 131:356-369.
[20] WU Y, ZHAO Y, YANG H Z, et al. HMGB1 regulates ferroptosis through Nrf2 pathway in mesangial cells in response to high glucose[J]. Biosci Rep, 2021, 41(2):BSR20202924.
[21] YANG W S, SRIRAMARATNAM R, WELSCH M E, et al. Regulation of ferroptotic cancer cell death by GPX4[J]. Cell, 2014, 156(1/2):317-331.
[22] SHEN E, FAN J, CHEN R Z, et al. Phospholipase Cγ1 signalling regulates lipopolysaccharide-induced cyclooxygenase-2 expression in cardiomyocytes[J]. J Mol Cell Cardiol, 2007, 43(3):308-318.
[23] MENG Z, LI H Y, SI C Y, et al. Asiatic acid inhibits cardiac fibrosis throughNrf2/HO-1 and TGF-β1/Smads signaling pathways in spontaneous hypertension rats[J]. Int Immunopharmacol, 2019, 74:105712.
[24] QIU W Y, ZHANG X T, PANG X Y, et al. Asiatic acid alleviates LPS-induced acute kidney injury in broilers by inhibiting oxidative stress and ferroptosis via activation of the Nrf2 pathway[J]. Food Chem Toxicol, 2022, 170:113468.
[25] QIU W Y, SU Y M, YE J L, et al. Study on Asiatic acid alleviates LPS-induced acute kidney injury by regulating apoptosis and autophagy of broilers[J/OL]. Acta Veterinaria et Zootechnica Sinica, 1-15[2023-12-28]. http://kns.cnki.net/kcms/detail/11.1985.S.20230928.0837.002.html. (in Chinese)
邱文粤, 苏依曼, 叶嘉莉, 等. 积雪草酸通过调控细胞凋亡和自噬缓解脂多糖诱导肉鸡急性肾损伤的研究[J/OL]. 畜牧兽医学报, 1-15[2023-12-28]. http://kns.cnki.net/kcms/detail/11.1985.S.20230928.0837.002.html.
[26] ATEYA A I, ARAFAT N, SALEH R M, et al. Intestinal gene expressions in broiler chickens infected with Escherichia coli and dietary supplemented with probiotic, acidifier and synbiotic[J]. Vet Res Commun, 2019, 43(2):131-142.
[27] SUZUKI K, YOSSAPOL M, SUGIYAMA M, et al. Effects of antimicrobial administration on the prevalence of antimicrobial-resistant Escherichia coli in broiler flocks[J]. Jpn J Infect Dis, 2019, 72(3):179-184.
[28] BAI T, HU X Y, ZHENG Y, et al. Resveratrol protects against lipopolysaccharide-induced cardiac dysfunction by enhancing SERCA2a activity through promoting the phospholamban oligomerization[J]. Am J Physiol Heart Circ Physiol, 2016, 311(4):H1051-H1062.
[29] SUN M, WANG R, HAN Q H. Inhibition of leukotriene B4 receptor 1 attenuates lipopolysaccharide-induced cardiac dysfunction:role of AMPK-regulated mitochondrial function[J]. Sci Rep, 2017, 7:44352.
[30] LU Y P, KAN H W, WANG Y, et al. Asiatic acid ameliorates hepatic ischemia/reperfusion injury in rats via mitochondria-targeted protective mechanism[J]. Toxicol Appl Pharmacol, 2018, 338:214-223.
[31] DONG S H, LIU Y W, WEI F, et al. Asiatic acid ameliorates pulmonary fibrosis induced by bleomycin (BLM) via suppressing pro-fibrotic and inflammatory signaling pathways[J]. Biomed Pharmacother, 2017, 89:1297-1309.
[32] PARK J H, SEO Y H, JANG J H, et al. Asiatic acid attenuates methamphetamine-induced neuroinflammation and neurotoxicity through blocking of NF-κB/STAT3/ERK and mitochondria-mediated apoptosis pathway[J]. J Neuroinflammation, 2017, 14(1):240.
[33] MA Z G, DAI J, WEI W Y, et al. Asiatic acid protects against cardiac hypertrophy through activating AMPKα signalling pathway[J]. Int J Biol Sci, 2016, 12(7):861-871.
[34] CAO W, LI X Q, ZHANG X N, et al. Madecassoside suppresses LPS-induced TNF-α production in cardiomyocytes through inhibition of ERK, p38, and NF-κB activity[J]. Int Immunopharmacol, 2010, 10(7):723-729.
[35] WEI S S, XIAO Z J, HUANG J, et al. Disulfiram inhibits oxidative stress and NLRP3 inflammasome activation to prevent LPS-induced cardiac injury[J]. Int Immunopharmacol, 2022, 105:108545.
[36] SONG Y X, OU Y M, ZHOU J Y. Gracillin inhibits apoptosis and inflammation induced by lipopolysaccharide (LPS) to alleviate cardiac injury in mice via improving miR-29a[J]. Biochem Biophys Res Commun, 2020, 523(3):580-587.
[37] QIU W Y, PANG X Y, ZHANG X T, et al. Protective effect of asiatic acid on LPS induced acute liver injury in broilers[J]. Chinese Journal of Veterinary Science, 2023, 43(1):164-171. (in Chinese)
邱文粤, 庞晓玥, 章心婷, 等. 积雪草酸对LPS诱导肉鸡急性肝损伤的保护作用[J]. 中国兽医学报, 2023, 43(1):164-171.
[38] ZHOU Y X, HU X F, ZHONG S W, et al. Effects of continuous LPS induction on oxidative stress and liver injury in weaned piglets[J]. Vet Sci, 2022, 10(1):22.
[39] JIANG S Q, CHEN Z L, ZHANG S, et al. Protective effects of protocatechuic acid on growth performance, intestinal barrier and antioxidant capacity in broilers challenged with lipopolysaccharide[J]. Animal, 2023, 17(1):100693.
[40] MA Q. Role of nrf2 in oxidative stress and toxicity[J]. Annu Rev Pharmacol Toxicol, 2013, 53:401-426.
[41] PRONETH B, CONRAD M. Ferroptosis and necroinflammation, a yet poorly explored link[J]. Cell Death Differ, 2019, 26(1):14-24.
[42] LANGSTON J W, LI W, HARRISON L, et al. Activation of promoter activity of the catalytic subunit of γ-glutamylcysteine ligase (GCL) in brain endothelial cells by insulin requires antioxidant response element 4 and altered glycemic status:implication for GCL expression and GSH synthesis[J]. Free Radic Biol Med, 2011, 51(9):1749-1757.
[43] SCHAUPP C M, BOTTA D, WHITE C C, et al. Persistence of improved glucose homeostasis in Gclm null mice with age and cadmium treatment[J]. Redox Biol, 2022, 49:102213.
[44] FANG X X, WANG H, HAN D, et al. Ferroptosis as a target for protection against cardiomyopathy[J]. Proc Natl Acad Sci U S A, 2019, 116(7):2672-2680.
[45] LI N, WANG W, ZHOU H, et al. Ferritinophagy-mediated ferroptosis is involved in sepsis-induced cardiac injury[J]. Free Radic Biol Med, 2020, 160:303-318.
[46] INGOLD I, BERNDT C, SCHMITT S, et al. Selenium utilization by GPX4 is required to prevent hydroperoxide-induced ferroptosis[J]. Cell, 2018, 172(3):409-422.e21.
[47] WANG Y, CHEN Q, SHI C X, et al. Mechanism of glycyrrhizin on ferroptosis during acute liver failure by inhibiting oxidative stress[J]. Mol Med Rep, 2019, 20(5):4081-4090.
[48] LO M, LING V, WANG Y Z, et al. The x_c^- cystine/glutamate antiporter:a mediator of pancreatic cancer growth with a role in drug resistance[J]. Br J Cancer, 2008, 99(3):464-472.
[49] GOZZELINO R, SOARES M P. Coupling heme and iron metabolism via ferritin H chain[J]. Antioxid Redox Signal, 2014, 20(11):1754-1769.

Effect of Asiatic Acid Alleviating Myocardial Injury Caused by Lipopolysaccharide through Inhibiting Oxidative Stress and Ferroptosis in Broilers

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