Nimbin, a bioactive triterpenoid compound isolated from the neem tree (Azadirachta indica), is known for its anti-inflammatory, antioxidant, antimicrobial, and hepatoprotective properties. The study aimed to explore the impact of nimbin on cardiac markers, mitochondrial and lysosomal enzyme activities, as well as apoptotic signaling in rats induced with isoproterenol. The subcutaneous injection of isoproterenol (ISO) at a dosage of 85 mg/kg body weight over the last two consecutive days led to notable increased in the activities/levels of the cardiac markers, lysosomal glycohydrolases and cathepsins. Conversely, reductions in the functioning of mitochondrial tricarboxylic acid cycle enzymes and respiratory chain enzymes in ISO-induced rats. In ISO-induced rats, there was an augmentation in the expressions of Bax, caspase-3, caspase-9, and cytochrome c, along with a diminished level of Bcl-2. Administration of nimbin resulted in decreased activities/levels cardiac markers, lysosomal glycohydrolases, cathepsins and increased functioning of mitochondrial tricarboxylic acid cycle enzymes and respiratory chain enzymes. Additionally, decreased expressions of Bax, executioner caspases and cytochrome c, along with heightened expression of Bcl-2, were noted in rats treated with nimbin. This indicates that nimbin possesses cardioprotective properties and mitigates mitochondrial and lysosomal dysfunction in rats induced with ISO. Key words Myocardial infarction " Tricarboxylic acid (TCA) cycle enzymes " Cathepsin-D " Cardiac markers " Triterpenoid.

Department of Cardiovascular Medicine 1st People's Hospital of Linping District Hangzhou Zhejiang China Department of Nephrology Xiangyang Central Hospital Affiliated Hospital of Hubei University of Arts and Science Xiangfan Hubei China

Zobrazit více v PubMed

Goldsborough E, 3rd, Tasdighi E, Blaha MJ. Assessment of cardiovascular disease risk: a 2023 update. Curr Opin Lipidol. 2023;34:162–173. doi: 10.1097/MOL.0000000000000887. PubMed DOI

Akila P, Asaikumar L, Vennila L. Chlorogenic acid ameliorates isoproterenol-induced myocardial injury in rats by stabilizing mitochondrial and lysosomal enzymes. Biomed Pharmacother. 2017;85:582–591. doi: 10.1016/j.biopha.2016.11.067. PubMed DOI

Anderson JL, Morrow DA. Acute Myocardial Infarction. N Engl J Med. 2017;376:2053–2064. doi: 10.1056/NEJMra1606915. PubMed DOI

Thangaiyan R, Arjunan S, Govindasamy K, Khan HA, Alhomida AS, Prasad NR. Galangin Attenuates Isoproterenol-Induced Inflammation and Fibrosis in the Cardiac Tissue of Albino Wistar Rats. Front Pharmacol. 2020;11:585163. doi: 10.3389/fphar.2020.585163. PubMed DOI PMC

Mohanty I, Arya DS, Dinda A, Talwar KK, Joshi S, Gupta SK. Mechanisms of cardioprotective effect of Withania somnifera in experimentally induced myocardial infarction. Basic Clin Pharmacol Toxicol. 2004;94:184–190. doi: 10.1111/j.1742-7843.2004.pto940405.x. PubMed DOI

Wong ZW, Thanikachalam PV, Ramamurthy S. Molecular understanding of the protective role of natural products on isoproterenol-induced myocardial infarction: A review. Biomed Pharmacother. 2017;94:1145–1166. doi: 10.1016/j.biopha.2017.08.009. PubMed DOI

Rathod S, Agrawal Y, Sherikar A, Nakhate KT, Patil CR, Nagoor Meeran MF, Ojha S, Goyal SN. Thymoquinone Produces Cardioprotective Effect in β-Receptor Stimulated Myocardial Infarcted Rats via Subsiding Oxidative Stress and Inflammation. Nutrients. 2022;14:2742. doi: 10.3390/nu14132742. PubMed DOI PMC

Khalil MI, Ahmmed I, Ahmed R, Tanvir EM, Afroz R, Paul S, Gan SH, Alam N. Amelioration of Isoproterenol-Induced Oxidative Damage in Rat Myocardium by Withania somnifera Leaf Extract. Biomed Res Int. 2015;2015:624159. doi: 10.1155/2015/624159. PubMed DOI PMC

Ferko M, Alanova P, Janko D, Opletalova B, Andelova N. Mitochondrial Peroxiredoxins and Monoamine Oxidase-A: Dynamic Regulators of ROS Signaling in Cardioprotection. Physiol Res. 2024;73:887–900. doi: 10.33549/physiolres.935513. PubMed DOI PMC

Huang Q, Chen Y. NADPH oxidase 4 contributes to oxidative stress in a mouse model of myocardial infarction. Physiol Res. 2023;72:177–186. doi: 10.33549/physiolres.934992. PubMed DOI PMC

Kumaran KS, Prince PS. Preventive effect of caffeic acid on lysosomal dysfunction in isoproterenol-induced myocardial infarcted rats. J Biochem Mol Toxicol. 2010;24:115–122. doi: 10.1002/jbt.20319. PubMed DOI

Decker RS, Poole AR, Griffin EE, Dingle JT, Wildenthal K. Altered distribution of lysosomal cathepsin D in ischemic myocardium. J Clin Invest. 1977;59:911–921. doi: 10.1172/JCI108713. PubMed DOI PMC

Sharma R, Kaushik S, Shyam H, Agarwal S, Balapure AK. Neem Seed Oil Induces Apoptosis in MCF-7 and MDA MB-231 Human Breast Cancer Cells. Asian Pac J Cancer Prev. 2017;18:2135–2140. doi: 10.22034/APJCP.2017.18.8.2135. PubMed DOI PMC

Charles V, Charles SX. The use efficacy of Azadirachta indica ADR (‘Neem’) Curcuma longa (‘Turmeric’) in scabies A pilot study. Trop Geogr Med. 1992;44:178–181. PubMed

Bandyopadhyay U, Biswas K, Sengupta A, Moitra P, Dutta P, Sarkar D, Debnath P, Ganguly CK, Banerjee RK. Clinical studies on the effect of Neem (Azadirachta indica) bark extract on gastric secretion and gastroduodenal ulcer. Life Sci. 2004;75:2867–2878. doi: 10.1016/j.lfs.2004.04.050. PubMed DOI

Bose A, Haque E, Baral R. Neem leaf preparation induces apoptosis of tumor cells by releasing cytotoxic cytokines from human peripheral blood mononuclear cells. Phytother Res. 2007;21:914–920. doi: 10.1002/ptr.2185. PubMed DOI

Kochhar A, Sharma N, Sachdeva R. Effect of supplementation of Tulsi (Ocimum sanctum) and Neem (Azadirachta indica) leaf powder on diabetic symptoms, anthropometric parameters and blood pressure of non-insulin dependent male diabetics. Studies Ethno Medicine. 2009;3:5–9. doi: 10.1080/09735070.2009.11886330. DOI

Paul R, Prasad M, Sah NK. Anticancer biology of Azadirachta indica L (neem): a mini review. Cancer Biol Ther. 2011;12:467–476. doi: 10.4161/cbt.12.6.16850. PubMed DOI

Nivetha R, Arvindh S, Baba AB, Gade DR, Gopal G, KC, Reddy KP, et al. Nimbolide, a Neem Limonoid, Inhibits Angiogenesis in Breast Cancer by Abrogating Aldose Reductase Mediated IGF-1/PI3K/Akt Signalling. Anticancer Agents Med Chem. 2022;22:2619–2636. doi: 10.2174/1871520622666220204115151. PubMed DOI

Mittal R, Chaudhry N, Mukherjee TK. Targeting breast cancer cell signaling molecules PI3K and Akt by phytochemicals Cannabidiol, Nimbin and Acetogenin: An in silico approach. J Biomed. 2018;3:60–63. doi: 10.7150/jbm.25815. DOI

Sharma V, Vijay J, Ganesh MR, Sundaramurthy A. Multilayer capsules encapsulating nimbin and doxorubicin for cancer chemo-photothermal therapy. Int J Pharm. 2020;582:119350. doi: 10.1016/j.ijpharm.2020.119350. PubMed DOI

Sudhakaran G, Rajesh R, Guru A, Arasu MV, Gopinath P, Arockiaraj J. Nimbin analogs N5 and N7 regulate the expression of lipid metabolic genes and inhibit lipid accumulation in high-fat diet-induced zebrafish larvae: An antihyperlipidemic study. Tissue Cell. 2023;80:102000. doi: 10.1016/j.tice.2022.102000. PubMed DOI

Sudhakaran G, Velayutham M, Aljarba NH, Al-Hazani TM, Arokiyaraj S, Guru A, Arockiaraj J. Nimbin (N1) and analog N3 from the neem seeds suppress the migration of osteosarcoma MG-63 cells and arrest the cells in a quiescent state mediated via activation of the caspase-modulated apoptotic pathway. Mol Biol Rep. 2023;50:7357–7369. doi: 10.1007/s11033-023-08627-7. PubMed DOI

Bell JL, Baron DN. A colorimetric method for determination of isocitrate dehydrogenase. Clinica Chimica Acta. 1960;5:740–747. doi: 10.1016/0009-8981(60)90017-6. DOI

Slater EC, Borner WD., Jr The effect of fluoride on the succinic oxidase system. Biochem J. 1952;52:185–196. doi: 10.1042/bj0520185. PubMed DOI PMC

Mehler AH, Kornberg A, Grisolia S, Ochoa S. The enzymatic mechanism of oxidation-reductions between malate or isocitrate and pyruvate. J Biol Chem. 1948;74:961–977. doi: 10.1016/S0021-9258(18)57306-3. PubMed DOI

Steginsky CA, Gruys KJ, Frey PA. alpha-Ketoglutarate dehydrogenase complex of Escherichia coli. A hybrid complex containing pyruvate dehydrogenase subunits from pyruvate dehydrogenase complex. J Biol Chem. 1985;260:13690–31369. doi: 10.1016/S0021-9258(17)38780-X. PubMed DOI

Pearl W, Cascarano J, Zweifach BW. Microdetermination of cytochrome oxidase in rat tissues by the oxidation on N-phenyl-p-phenylene diamine or ascorbic acid. J Histochem Cytochem. 1963;11:102–104. doi: 10.1177/11.1.102. DOI

Minakami S, Ringler RL, Singer TP. Studies on the respiratory chain-linked dihydrodiphosphopyridine nucleotide dehydrogenase I. Assay of the enzyme in particulate and in soluble preparation. J Biol Chem. 1962;237:569–576. doi: 10.1016/S0021-9258(18)93963-3. PubMed DOI

DePierre JW, Karnovsky ML. Plasma membranes of mammalian cells: a review of methods for their characterization and isolation. J Cell Biol. 1973;56:275–303. doi: 10.1083/jcb.56.2.275. PubMed DOI PMC

Van Hoof F, Hers HG. The abnormalities of lysosomal enzymes in mucopolysaccharidoses. Eur J Biochem. 1968;7:34–44. doi: 10.1111/j.1432-1033.1968.tb19570.x. PubMed DOI

Conchie J, Gelman AL, Levvy GA. Inhibition of glycosidases by aldonolactones of corresponding configuration. The C-4- and C-6-specificity of beta-glucosidase and beta-galactosidase. Biochem J. 1967;103:609–615. doi: 10.1042/bj1030609. PubMed DOI PMC

Kawai Y, Anno K. Mucopolysaccharide-degrading enzymes from the liver of the squid, Ommastrephes sloani pacificus I. Hyaluronidase. Biochim Biophys Acta. 1971;242:428–436. doi: 10.1016/0005-2744(71)90234-8. PubMed DOI

Moore JC, Morris JE. A simple automated colorimetric method for determination of N-acetyl-beta-D-glucosaminidase. Ann Clin Biochem. 1982;19:157–159. doi: 10.1177/000456328201900305. PubMed DOI

Barrett AJ. A new assay for cathepsin B1 and other thiol proteinases. Anal Biochem. 1972;47:280–293. doi: 10.1016/0003-2697(72)90302-8. PubMed DOI

Sapolsky AI, Altman RD, Howell DS. Cathepsin D activity in normal and osteoarthritic human cartilage. Fed Proc. 1973;32:1489–1493. PubMed

Lobo Filho HG, Ferreira NL, Sousa RB, Carvalho ER, Lobo PL, Lobo Filho JG. Experimental model of myocardial infarction induced by isoproterenol in rats. (Article in English, Portuguese) Rev Bras Cir Cardiovasc. 2011;26:469–476. doi: 10.5935/1678-9741.20110024. PubMed DOI

Houston M. The role of nutrition and nutraceutical supplements in the treatment of hypertension. World J Cardiol. 2014;6:38–66. doi: 10.4330/wjc.v6.i2.38. PubMed DOI PMC

Wang SB, Tian S, Yang F, Yang HG, Yang XY, Du GH. Cardioprotective effect of salvianolic acid A on isoproterenol-induced myocardial infarction in rats. Eur J Pharmacol. 2009;615:125–132. doi: 10.1016/j.ejphar.2009.04.061. PubMed DOI

Gan ZH, Cheong HC, Tu YK, Kuo PH. Association between Plant-Based Dietary Patterns and Risk of Cardiovascular Disease: A Systematic Review and Meta-Analysis of Prospective Cohort Studies. Nutrients. 2021;13:3952. doi: 10.3390/nu13113952. PubMed DOI PMC

Gupta SC, Prasad S, Tyagi AK, Kunnumakkara AB, Aggarwal BB. Neem (Azadirachta indica): An indian traditional panacea with modern molecular basis. Phytomedicine. 2017;34:14–20. doi: 10.1016/j.phymed.2017.07.001. PubMed DOI

Chen W, Liang J, Fu Y, Jin Y, Yan R, Chi J, Liu W, Liu Y, Yin X. Cardioprotection of cortistatin against isoproterenol-induced myocardial injury in rats. Ann Transl Med. 2020;8:309. doi: 10.21037/atm.2020.02.93. PubMed DOI PMC

Ouyang B, Li Z, Ji X, Huang J, Zhang H, Jiang C. The protective role of lutein on isoproterenol-induced cardiac failure rat model through improving cardiac morphology, antioxidant status via positively regulating Nrf2/HO-1 signalling pathway. Pharm Biol. 2019;57:529–535. doi: 10.1080/13880209.2019.1649436. PubMed DOI PMC

Radhiga T, Rajamanickam C, Senthil S, Pugalendi KV. Effect of ursolic acid on cardiac marker enzymes, lipid profile and macroscopic enzyme mapping assay in isoproterenol-induced myocardial ischemic rats. Food Chem Toxicol. 2012;50:3971–3977. doi: 10.1016/j.fct.2012.07.067. PubMed DOI

Park KC, Gaze DC, Collinson PO, Marber MS. Cardiac troponins: from myocardial infarction to chronic disease. Cardiovasc Res. 2017;113:1708–1718. doi: 10.1093/cvr/cvx183. PubMed DOI PMC

Kumar M, Kasala ER, Bodduluru LN, Dahiya V, Lahkar M. Baicalein protects isoproterenol induced myocardial ischemic injury in male Wistar rats by mitigating oxidative stress and inflammation. Inflamm Res. 2016;65:613–622. doi: 10.1007/s00011-016-0944-z. PubMed DOI

Finsterer J, Ohnsorge P. Influence of mitochondrion-toxic agents on the cardiovascular system. Regul Toxicol Pharmacol. 2013;67:434–445. doi: 10.1016/j.yrtph.2013.09.002. PubMed DOI

Matsuzaki S, Szweda PA, Szweda LI, Humphries KM. Regulated production of free radicals by the mitochondrial electron transport chain: Cardiac ischemic preconditioning. Adv Drug Deliv Rev. 2009;61:1324–1331. doi: 10.1016/j.addr.2009.05.008. PubMed DOI PMC

Radhiga T, Senthil S, Sundaresan A, Pugalendi KV. Ursolic acid modulates MMPs, collagen-I, α-SMA, and TGF-β expression in isoproterenol-induced myocardial infarction in rats. Hum Exp Toxicol. 2019;38:785–793. doi: 10.1177/0960327119842620. PubMed DOI

Bhat OM, Li PL. Lysosome Function in Cardiovascular Diseases. Cell Physiol Biochem. 202;55:277–300. doi: 10.33594/000000373. PubMed DOI PMC

Chi C, Riching AS, Song K. Lysosomal Abnormalities in Cardiovascular Disease. Int J Mol Sci. 2020;21:811. doi: 10.3390/ijms21030811. PubMed DOI PMC

Kurian GA, Rajagopal R, Vedantham S, Rajesh M. The Role of Oxidative Stress in Myocardial Ischemia and Reperfusion Injury and Remodeling: Revisited. Oxid Med Cell Longev. 2016;2016:1656450. doi: 10.1155/2016/1656450. PubMed DOI PMC

Redza-Dutordoir M, Averill-Bates DA. Activation of apoptosis signalling pathways by reactive oxygen species. Biochim Biophys Acta. 2016;1863:2977–2992. doi: 10.1016/j.bbamcr.2016.09.012. PubMed DOI

Kale J, Osterlund EJ, Andrews DW. BCL-2 family proteins: changing partners in the dance towards death. Cell Death Differ. 2018;25:65–80. doi: 10.1038/cdd.2017.186. PubMed DOI PMC

Chen Z, Wu J, Li S, Liu C, Ren Y. Inhibition of Myocardial Cell Apoptosis Is Important Mechanism for Ginsenoside in the Limitation of Myocardial Ischemia/Reperfusion Injury. Front Pharmacol. 2022;13:806216. doi: 10.3389/fphar.2022.806216. PubMed DOI PMC