Emodin promotes autophagy to improve myocardial injury in septic model mice

doi:10.12307/2025.771

Abstract

Abstract: BACKGROUND: Emodin has a variety of pharmacological activities such as anti-inflammatory, anti-viral and anti-oxidative stress, and also has a certain protective effect on sepsis-induced myocardial injury, but its mechanism of action is still unclear.
OBJECTIVE: To investigate whether emodin can improve myocardial injury in septic mice by promoting autophagy.
METHODS: Thirty-two male Kunming mice were divided into sham operation group (n=4), sham operation+emodin group (n=4), model group (n=8), model+emodin group (n=8), and emodin+3-methyladenine group (n=8). The myocardial injury model of septic mice was constructed by cecal ligation and puncture. 3-methyladenine (10 mg/kg) was injected intraperitoneally 1 hour before modeling. Emodin (20 mg/kg) was injected intraperitoneally 30 minutes before modeling, and the other groups were injected with the same amount of normal saline at the same time point. Blood and myocardial samples were collected from all mice 24 hours after surgery. ELISA was used to detect the levels of brain natriuretic peptide and cardiac troponin I in serum. Western blot assay was used to detect the protein expression of LC3B, Beclin-1, and p62 in myocardial tissue. Hematoxylin-eosin staining was used to observe the pathological changes in myocardial tissue. Ultrasound was used to evaluate the cardiac function of mice.
RESULTS AND CONCLUSION: (1) Compared with the sham operation group, there was no significant difference in the levels of serum brain natriuretic peptide, cardiac troponin I, and the protein expression of myocardial autophagy proteins LC3II/LC3I and p62 in the sham operation+emodin group (P > 0.05). (2) Compared with the sham operation+emodin group, the levels of serum brain natriuretic peptide and cardiac troponin I were significantly increased in the model group (P < 0.05). Compared with the model group, the levels of serum brain natriuretic peptide and cardiac troponin I were decreased in the model+emodin group (P < 0.05). (3) Compared with the model group, the expression of LC3II/LC3I and Beclin-1 protein was increased and the expression of p62 protein was decreased in the myocardial tissue of the model+emodin group (P < 0.05). Compared with the model+emodin group, the expression of LC3II/LC3I and Beclin-1 protein decreased and the expression of p62 protein increased in the emodin+3-methyladenine group (P < 0.05). (4) The myocardial fibers in the sham operation group were normal, the myocardial fibers in the model group were disordered with a large number of inflammatory cell infiltration, the myocardial fibers in the model+emodin group were slightly disordered, and some vacuolar changes were observed. The myocardial fibers were disordered, and more inflammatory cell infiltration was observed in the emodin+3-methyladenine group. (5) Compared with the sham operation group, the left ventricular short axis shortening rate and left ventricular ejection fraction were decreased in the model group (P < 0.05). Compared with the model group, the left ventricular short axis shortening rate and left ventricular ejection fraction were increased in the model+emodin group (P < 0.05). Compared with the model+emodin group, the left ventricular ejection fraction of emodin+3-methyladenine group was decreased (P < 0.05), and the left ventricular short axis shortening rate was reduced but not statistically significant (P > 0.05). (6) The above results indicate that emodin pretreatment can improve myocardial injury and myocardial dysfunction in septic mice by promoting autophagy.

Key words: emodin, sepsis, sepsis-induced myocardial injury, cardiac function, autophagy, inflammatory reaction, protective effect, engineered tissue construction

CLC Number:

Tian Yong, Zhou Qing, Luo Chuanquan, Hu Hongmei, Ma Changlin, Yang Lei, Wei Lin. Emodin promotes autophagy to improve myocardial injury in septic model mice[J]. Chinese Journal of Tissue Engineering Research, 2025, 29(26): 5572-5578.

Figures/Tables 6

2.2 大黄素预处理对脓毒症小鼠心肌损伤的保护作用采用ELISA方法评估各组小鼠血清中代表心脏损伤程度的生物标志物水平(图1)。与假手术组相比，假手术+大黄素组脑钠肽、心肌肌钙蛋白I水平均无统计学意义(P > 0.05)；与假手术+大黄素组相比，模型组脑钠肽、心肌肌钙蛋白I水平均显著升高(P < 0.05)；与模型组相比，模型+大黄素组脑钠肽、心肌肌钙蛋白I水平均显著降低(P < 0.05)。 2.3 大黄素预处理对脓毒症小鼠心肌损伤自噬水平的影响采用Western blot方法检测各组小鼠心肌组织中自噬相关蛋白LC3B与p62的相对表达量(图2)，与假手术组相比，假手术+大黄素组和模型组自噬标志物LC3Ⅱ/LC3Ⅰ、p62的蛋白相对表达量差异无显著性意义(P > 0.05)；与模型组相比，模型+大黄素组LC3Ⅱ/LC3Ⅰ的蛋白相对表达量显著升高且p62的蛋白相对表达量显著降低(P < 0.05)。 2.4 3-甲基腺嘌呤对脓毒症小鼠心肌损伤自噬水平的作用采用Western blot方法检测各组小鼠心肌中自噬蛋白LC3B、Beclin-1及p62的相对表达水平(图3)，与假手术组相比，模型组LC3Ⅱ/LC3Ⅰ、Beclin-1及p62的蛋白相对表达量差异无显著性意义(P > 0.05)；与模型组相比，模型+大黄素组LC3Ⅱ/LC3Ⅰ和Beclin-1的蛋白相对表达量均显著升高且p62的蛋白相对表达量显著降低(P < 0.05)；与模型+大黄素组相比，大黄素+3-甲基腺嘌呤组LC3Ⅱ/LC3Ⅰ和Beclin-1的蛋白相对表达量均显著降低且p62的蛋白相对表达量显著升高(P < 0.05)。 2.5 大黄素促进自噬对脓毒症小鼠心肌损伤的作用采用苏木精-伊红染色观察各组小鼠心肌组织的病理变化特征(图4)。假手术组小鼠心肌纤维结构有序，无明显水肿迹象，亦未观察到炎症细胞的浸润；模型组小鼠展现出心肌纤维排列杂乱、伴随显著组织水肿及大量炎症细胞的浸润现象；模型+大黄素组心肌纤维排列轻微紊乱、轻度水肿，部分组织出现少量空泡样结构变化；大黄素+3-甲基腺嘌呤组心肌组织表现出结构紊乱、水肿，并见较多的炎症细胞浸润。 2.6 大黄素促进自噬对脓毒症小鼠心肌功能障碍的作用采用动物超声技术评估各组小鼠的左心室短轴缩短率及左心室射血分数(图5)。与假手术组相比，模型组左心室短轴缩短率、左心室射血分数均显著降低(P < 0.05)；与模型组相比，模型+大黄素组左室短轴缩短率、左室射血分数均升高(P < 0.05)；与模型+大黄素组相比，大黄素+3-甲基腺嘌呤组左室射血分数降低(P < 0.05)，左室短轴缩短率降低但无统计学意义(P > 0.05)。"

References

[1] PÓVOA P, COELHO L, DAL-PIZZOL F, et al. How to use biomarkers of infection or sepsis at the bedside: guide to clinicians. Intensive Care Med. 2023;49(2):142-153.
[2] LI Y, ZHANG L, ZHANG P, et al. Dehydrocorydaline Protects Against Sepsis-Induced Myocardial Injury Through Modulating the TRAF6/NF-κB Pathway. Front Pharmacol. 2021;12:709604.
[3] WANG Z, QIANG X, PENG Y, et al. Design and synthesis of salidroside analogs and their bioactivity against septic myocardial injury. Bioorg Chem. 2023;138:106609.
[4] FRENCKEN JF, VAN SMEDEN M, VAN DE GROEP K, et al. Etiology of Myocardial Injury in Critically Ill Patients with Sepsis: A Cohort Study. Ann Am Thorac Soc. 2022;19(5):773-780.
[5] BI CF, LIU J, YANG LS, et al. Research Progress on the Mechanism of Sepsis Induced Myocardial Injury. J Inflamm Res. 2022;15:4275-4290.
[6] GAO Y, LIU J, LI K, et al. Metformin Alleviates Sepsis-Associated Myocardial Injury by Enhancing AMP-Activated Protein Kinase/Mammalian Target of Rapamycin Signaling Pathway-Mediated Autophagy. J Cardiovasc Pharmacol. 2023;82(4):308-317.
[7] KLIONSKY DJ, PETRONI G, AMARAVADI RK, et al. Autophagy in major human diseases. EMBO J. 2021;40(19):e108863.
[8] SUN Y, CAI Y, ZANG QS. Cardiac Autophagy in Sepsis. Cells. 2019;8(2): 141.
[9] CAO W, LI J, YANG K, et al. An overview of autophagy: Mechanism, regulation and research progress. Bull Cancer. 2021;108(3):304-322.
[10] IRIONDO MN, ETXANIZ A, VARELA YR, et al. LC3 subfamily in cardiolipin-mediated mitophagy: a comparison of the LC3A, LC3B and LC3C homologs. Autophagy. 2022;18(12):2985-3003.
[11] SUN Y, YAO X, ZHANG QJ, et al. Beclin-1-Dependent Autophagy Protects the Heart During Sepsis. Circulation. 2018;138(20):2247-2262.
[12] SHU X, SUN Y, SUN X, et al. The effect of fluoxetine on astrocyte autophagy flux and injured mitochondria clearance in a mouse model of depression. Cell Death Dis. 2019;10(8):577.
[13] HO J, YU J, WONG SH, et al. Autophagy in sepsis: Degradation into exhaustion? Autophagy. 2016;12(7):1073-1082.
[14] ZHANG P, LI Y, FU Y, et al. Inhibition of Autophagy Signaling via 3-methyladenine Rescued Nicotine-Mediated Cardiac Pathological Effects and Heart Dysfunctions. Int J Biol Sci. 2020;16(8):1349-1362.
[15] BARICHELLO T, GENEROSO JS, SINGER M, et al. Biomarkers for sepsis: more than just fever and leukocytosis-a narrative review. Crit Care. 2022;26(1):14.
[16] MCDONALD SJ, VANDERVEEN BN, VELAZQUEZ KT, et al. Therapeutic Potential of Emodin for Gastrointestinal Cancers. Integr Cancer Ther. 2022;21:15347354211067469.
[17] QIN B, ZENG Z, XU J, et al. Emodin inhibits invasion and migration of hepatocellular carcinoma cells via regulating autophagy-mediated degradation of snail and β-catenin. BMC Cancer. 2022;22(1):671.
[18] 徐瑞明,邵峥谊,王大为,等.大黄素对脓毒症大鼠心肌损伤的保护作用 [J].西部医学,2020,32(10):1443-1446.
[19] 田勇,周颖,古雍翔,等.二甲双胍预处理诱导心脏自噬减轻脓毒症小鼠的心肌损伤[J].中国组织工程研究,2024,28(28):4469-4476.
[20] GAO LL, WANG ZH, MU YH, et al. Emodin Promotes Autophagy and Prevents Apoptosis in Sepsis-Associated Encephalopathy through Activating BDNF/TrkB Signaling. Pathobiology. 2022;89(3):135-145.
[21] 田勇,周颖,古雍翔,等.二甲双胍诱导心肌细胞自噬对脓毒症小鼠心肌损伤的保护机制[J].安徽医科大学学报,2024,59(1):92-98.
[22] PEI XB, LIU B. Research Progress on the Mechanism and Management of Septic Cardiomyopathy: A Comprehensive Review. Emerg Med Int. 2023;2023:8107336.
[23] ZOU HX, QIU BQ, ZHANG ZY, et al. Dysregulated autophagy-related genes in septic cardiomyopathy: Comprehensive bioinformatics analysis based on the human transcriptomes and experimental validation. Front Cardiovasc Med. 2022;9:923066.
[24] TANG R, JIA L, LI Y, et al. Narciclasine attenuates sepsis-induced myocardial injury by modulating autophagy. Aging (Albany NY). 2021; 13(11):15151-15163.
[25] WONG SQ, KUMAR AV, MILLS J, et al. Autophagy in aging and longevity. Hum Genet. 2020;139(3):277-290.
[26] GROSS AS, GRAEf M. Mechanisms of Autophagy in Metabolic Stress Response. J Mol Biol. 2020;432(1):28-52.
[27] LIU C, LIU Y, CHEN H, et al. Myocardial injury: where inflammation and autophagy meet. Burns Trauma. 2023;11:tkac062.
[28] DU J, ZHOU Y. Propofol reduces lipopolysaccharide‑induced cardiomyocyte injury in sepsis by activating SIRT1‑mediated autophagy. Exp Ther Med. 2023;25(4):187.
[29] WANG X, XIE D, DAI H, et al. Clemastine protects against sepsis-induced myocardial injury in vivo and in vitro. Bioengineered. 2022;13(3): 7134-7146.
[30] QIN GW, LU P, PENG L, et al. Ginsenoside Rb1 Inhibits Cardiomyocyte Autophagy via PI3K/Akt/mTOR Signaling Pathway and Reduces Myocardial Ischemia/Reperfusion Injury. Am J Chin Med. 2021;49(8): 1913-1927.
[31] WANG X, YANG S, LI Y, et al. Role of emodin in atherosclerosis and other cardiovascular diseases: Pharmacological effects, mechanisms, and potential therapeutic target as a phytochemical. Biomed Pharmacother. 2023;161:114539.
[32] SU J, ZHOU F, WU S, et al. Research Progress on Natural Small-Molecule Compounds for the Prevention and Treatment of Sepsis. Int J Mol Sci. 2023;24(16):12732.
[33] CUI Y, CHEN LJ, HUANG T, et al. The pharmacology, toxicology and therapeutic potential of anthraquinone derivative emodin. Chin J Nat Med. 2020;18(6):425-435.
[34] DAI S, YE B, CHEN L, et al. Emodin alleviates LPS-induced myocardial injury through inhibition of NLRP3 inflammasome activation. Phytother Res. 2021;35(9):5203-5213.
[35] TU YJ, TAN B, JIANG L, et al. Emodin Inhibits Lipopolysaccharide-Induced Inflammation by Activating Autophagy in RAW 264.7 Cells. Chin J Integr Med. 2021;27(5):345-352.
[36] LIU H, WANG Q, SHI G, et al. Emodin Ameliorates Renal Damage and Podocyte Injury in a Rat Model of Diabetic Nephropathy via Regulating AMPK/mTOR-Mediated Autophagy Signaling Pathway. Diabetes Metab Syndr Obes. 2021;14:1253-1266.
[37] WU P, XIAO Y, QING L, et al. Emodin activates autophagy to suppress oxidative stress and pyroptosis via mTOR-ULK1 signaling pathway and promotes multi-territory perforator flap survival. Biochem Biophys Res Commun. 2024;704:149688.
[38] SHE H, TAN L, DU Y, et al. VDAC2 malonylation participates in sepsis-induced myocardial dysfunction via mitochondrial-related ferroptosis. Int J Biol Sci. 2023;19(10):3143-3158.
[39] SHEN XD, ZHANG HS, ZHANG R, et al. Progress in the Clinical Assessment and Treatment of Myocardial Depression in Critically Ill Patient with Sepsis. J Inflamm Res. 2022;15:5483-5490.
[40] LIU FJ, GU TJ, WEI DY. Emodin alleviates sepsis-mediated lung injury via inhibition and reduction of NF-kB and HMGB1 pathways mediated by SIRT1. Kaohsiung J Med Sci. 2022;38(3):253-260.
[41] GUO R, LI Y, HAN M, et al. Emodin attenuates acute lung injury in Cecal-ligation and puncture rats. Int Immunopharmacol. 2020;85:106626.
[42] DONG Y, ZHANG L, JIANG Y, et al. Emodin reactivated autophagy and alleviated inflammatory lung injury in mice with lethal endotoxemia. Exp Anim. 2019;68(4):559-568.
[43] CHANG X, HE Y, WANG L, et al. Puerarin Alleviates LPS-Induced H9C2 Cell Injury by Inducing Mitochondrial Autophagy. J Cardiovasc Pharmacol. 2022;80(4):600-608.
[44] ZHANG W, ZHANG J. Semaglutide Pretreatment Induces Cardiac Autophagy to Reduce Myocardial Injury in Septic Mice. Discov Med. 2023;35(178):853-860.
[45] WU B, SONG H, FAN M, et al. Luteolin attenuates sepsis‑induced myocardial injury by enhancing autophagy in mice. Int J Mol Med. 2020;45(5):1477-1487.