Abstract
To investigate the efficacy of Slit2 in the rats with coronary heart disease (CHD). CHD model were constructed by feeding high-fat food and injecting with pituitrin in rat, followed by recombinant Slit2 treatment, and then the cardiac function was evaluated by echocardiography, and the indicators concerning the cardiomyocyte injury markers and lipoprotein status and oxidative stress were measured. The Slit2 expression in the heart tissues was identified by immunofluorescence. Enzyme-linked immunosorbent assay (ELISA) was carried out to detect inflammatory cytokines, H2DCFDA staining to determine the ROS generation in heart tissues, Masson trichrome staining to observe myocardial fibrosis, and qRT-PCR and Western blotting to detect gene and protein expressions. Slit2 decreased the levels of LDH, CK-MB, cTnI, TG, TC and LDL-C and increased HDL-C level in CHD rats. In the normal heart tissues, Slit2 expression was significantly lower in cardiomyocytes than cardiac fibroblasts. Furthermore, the expressions of VCAM-1, ICAM-1, fibronectin and TGF-β1 were increased in the heart tissues of CHD rats with the obvious myocardial fibrosis, which were dose-dependently reversed by recombinant Slit2. In addition, recombinant Slit2 also dose-dependently increased the activity of NO, SOD, CAT and GSH-Px, and decreased TNF-α, IL-6, MCP-1, MDA and ROS in CHD rats. Slit2 was downregulated in myocardial tissue and plasma of CHD rats. Recombinant Slit2, by regulating the level of blood lipid, can relieve the myocardial fibrosis, inflammation and oxidative stress in CHD.
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Tu, S., Xiao, F., Min, X., Chen, H., Fan, X., & Cao, K. (2018). Catechin attenuates coronary heart disease in a rat model by inhibiting inflammation. Cardiovascular Toxicology, 18, 393–399.
Heusch, G., Schulz, R., & Erbel, R. (2003). Inflammatory markers in coronary heart disease: coronary vascular versus myocardial origin? Circulation, 108, e4.
Nusslein-Volhard, C., Wieschaus, E., & Kluding, H. (1984). Mutations affecting the pattern of the larval cuticle in Drosophila melanogaster: I. Zygotic loci on the second chromosome. Wilehm Roux’s Archives of Developmental Biology, 193, 267–282.
Qian, L., Liu, J., & Bodmer, R. (2005). Slit and Robo control cardiac cell polarity and morphogenesis. Current Biology, 15, 2271–2278.
Fish, J. E., Wythe, J. D., Xiao, T., Bruneau, B. G., Stainier, D. Y., Srivastava, D., & Woo, S. (2011). A Slit/miR-218/Robo regulatory loop is required during heart tube formation in zebrafish. Development, 138, 1409–1419.
Medioni, C., Bertrand, N., Mesbah, K., Hudry, B., Dupays, L., Wolstein, O., Washkowitz, A. J., Papaioannou, V. E., Mohun, T. J., Harvey, R. P., & Zaffran, S. (2010). Expression of Slit and Robo genes in the developing mouse heart. Developmental Dynamics, 239, 3303–3311.
Mommersteeg, M. T., Andrews, W. D., Ypsilanti, A. R., Zelina, P., Yeh, M. L., Norden, J., Kispert, A., Chedotal, A., Christoffels, V. M., & Parnavelas, J. G. (2013). Slit-roundabout signaling regulates the development of the cardiac systemic venous return and pericardium. Circulation Research, 112, 465–475.
Calmont, A., Ivins, S., Van Bueren, K. L., Papangeli, I., Kyriakopoulou, V., Andrews, W. D., Martin, J. F., Moon, A. M., Illingworth, E. A., Basson, M. A., & Scambler, P. J. (2009). Tbx1 controls cardiac neural crest cell migration during arch artery development by regulating Gbx2 expression in the pharyngeal ectoderm. Development, 136, 3173–3183.
Yu, H., Gao, G., Jiang, L., Guo, L., Lin, M., Jiao, X., Jia, W., & Huang, J. (2013). Decreased expression of miR-218 is associated with poor prognosis in patients with colorectal cancer. International Journal of Clinical and Experimental Pathology, 6, 2904–2911.
Nguyen Ba-Charvet, K. T., Brose, K., Ma, L., Wang, K. H., Marillat, V., Sotelo, C., Tessier-Lavigne, M., & Chedotal, A. (2001). Diversity and specificity of actions of Slit2 proteolytic fragments in axon guidance. Journal of Neuroscience, 21, 4281–4289.
Katoh, Y., & Katoh, M. (2005). Comparative genomics on SLIT1, SLIT2, and SLIT3 orthologs. Oncology Reports, 14, 1351–1355.
Pilling, D., Zheng, Z., Vakil, V., & Gomer, R. H. (2014). Fibroblasts secrete Slit2 to inhibit fibrocyte differentiation and fibrosis. Proceedings of the National Academy of Sciences of the United States of America, 111, 18291–18296.
Chang, J., Lan, T., Li, C., Ji, X., Zheng, L., Gou, H., Ou, Y., Wu, T., Qi, C., Zhang, Q., Li, J., Gu, Q., Wen, D., Cao, L., Qiao, L., Ding, Y., & Wang, L. (2015). Activation of Slit2-Robo1 signaling promotes liver fibrosis. Journal of Hepatology, 63, 1413–1420.
Guan, H., Zu, G., Xie, Y., Tang, H., Johnson, M., Xu, X., Kevil, C., Xiong, W. C., Elmets, C., Rao, Y., Wu, J. Y., & Xu, H. (2003). Neuronal repellent Slit2 inhibits dendritic cell migration and the development of immune responses. The Journal of Immunology, 171, 6519–6526.
Tole, S., Mukovozov, I. M., Huang, Y. W., Magalhaes, M. A., Yan, M., Crow, M. R., Liu, G. Y., Sun, C. X., Durocher, Y., Glogauer, M., & Robinson, L. A. (2009). The axonal repellent, Slit2, inhibits directional migration of circulating neutrophils. Journal of Leukocyte Biology, 86, 1403–1415.
Chaturvedi, S., Yuen, D. A., Bajwa, A., Huang, Y. W., Sokollik, C., Huang, L., Lam, G. Y., Tole, S., Liu, G. Y., Pan, J., Chan, L., Sokolskyy, Y., Puthia, M., Godaly, G., John, R., Wang, C., Lee, W. L., Brumell, J. H., Okusa, M. D., & Robinson, L. A. (2013). Slit2 prevents neutrophil recruitment and renal ischemia-reperfusion injury. Journal of the American Society of Nephrology, 24, 1274–1287.
Dallol, A., Krex, D., Hesson, L., Eng, C., Maher, E. R., & Latif, F. (2003). Frequent epigenetic inactivation of the SLIT2 gene in gliomas. Oncogene, 22, 4611–4616.
Kim, H. K., Zhang, H., Li, H., Wu, T. T., Swisher, S., He, D., Wu, L., Xu, J., Elmets, C. A., Athar, M., Xu, X. C., & Xu, H. (2008). Slit2 inhibits growth and metastasis of fibrosarcoma and squamous cell carcinoma. Neoplasia, 10, 1411–1420.
Chen, G. X., Wang, H. Y., Liu, T., Yang, M. T., Zhou, Z. Y., & Feng, G. (2013). Myocardial Slit2/Robo4 expression and impact of exogenous Slit2 on proliferation and migration of cardiac microvascular endothelial cells. Zhonghua Xin Xue Guan Bing Za Zhi, 41, 1034–1039.
Li, X., Zheng, S., Tan, W., Chen, H., Li, X., Wu, J., Luo, T., Ren, X., Pyle, W. G., Wang, L., Backx, P. H., Huang, R., & Yang, F. H. (2020). slit2 protects hearts against ischemia-reperfusion injury by inhibiting inflammatory responses and maintaining myofilament contractile properties. Frontiers in Physiology, 11, 228.
Carbone, L. (2012). Pain management standards in the Guide for the Care and Use of Laboratory Animals. Journal of the American Association for Laboratory Animal Science, 51, 322–328.
Goth, L. (1991). A simple method for determination of serum catalase activity and revision of reference range. Clinica Chimica Acta, 196, 143–151.
Sazuka, Y., Tanizawa, H., & Takino, Y. (1989). Effect of adriamycin on the activities of superoxide dismutase, glutathione peroxidase and catalase in tissues of mice. Japanese Journal of Cancer Research, 80, 89–94.
Xu, J., Zhou, X., Deng, Q., Huang, Q., Yang, J., & Huang, F. (2011). Rapeseed oil fortified with micronutrients reduces atherosclerosis risk factors in rats fed a high-fat diet. Lipids in Health and Disease, 10, 96.
Sung, P. H., Sun, C. K., Ko, S. F., Chang, L. T., Sheu, J. J., Lee, F. Y., Wu, C. J., Chua, S., & Yip, H. K. (2009). Impact of hyperglycemic control on left ventricular myocardium. A molecular and cellular basic study in a diabetic rat model. International Heart Journal, 50, 191–206.
Chang, H., Wang, Q., Shi, T., Huo, K., Li, C., Zhang, Q., Wang, G., Wang, Y., Tang, B., Wang, W., & Wang, Y. (2016). Effect of DanQi Pill on PPARalpha, lipid disorders and arachidonic acid pathway in rat model of coronary heart disease. BMC Complementary and Alternative Medicine, 16, 103.
Shu, J., Huang, R., Tian, Y., Liu, Y., Zhu, R., & Shi, G. (2020). Andrographolide protects against endothelial dysfunction and inflammatory response in rats with coronary heart disease by regulating PPAR and NF-kappaB signaling pathways. Annals of Palliative Medicine, 9, 1965–1975.
Steven, S., Frenis, K., Oelze, M., Kalinovic, S., Kuntic, M., Bayo Jimenez, M. T., Vujacic-Mirski, K., Helmstadter, J., Kroller-Schon, S., Munzel, T., & Daiber, A. (2019). Vascular inflammation and oxidative stress: major triggers for cardiovascular disease. Oxidative Medicine and Cellular Longevity, 2019, 7092151.
Lin, Y., Dan, H., & Lu, J. (2020). Overexpression of microRNA-136-3p alleviates myocardial injury in coronary artery disease via the Rho A/ROCK signaling pathway. Kidney & Blood Pressure Research, 45, 477–496.
Li, X., Lu, Y., Sun, Y., & Zhang, Q. (2015). Effect of curcumin on permeability of coronary artery and expression of related proteins in rat coronary atherosclerosis heart disease model. International Journal of Clinical and Experimental Pathology, 8, 7247–7253.
Yusuf, B., Mukovozov, I., Patel, S., Huang, Y. W., Liu, G. Y., Reddy, E. C., Skrtic, M., Glogauer, M., & Robinson, L. A. (2021). The neurorepellent, Slit2, prevents macrophage lipid loading by inhibiting CD36-dependent binding and internalization of oxidized low-density lipoprotein. Science and Reports, 11, 3614.
Lu, N., Du, Y., Li, H., Luo, Y., Ouyang, B., Chen, Y., Yang, Y., & Yang, L. (2018). Omega-6 fatty acids down-regulate matrix metalloproteinase expression in a coronary heart disease-induced rat model. International Journal of Experimental Pathology, 99, 210–217.
Cappola, T., Li, M., Jing, H., Ky, B., Gilmore, J., Keating, B., Reilly, M., Syed, F., Matkovich, S., & Gerald Dorn, I. (2008). Abstract 4874: Large-scale candidate gene association with human heart failure. Circulation, 103, 530.
Moreira, R. S., Irigoyen, M., Sanches, T. R., Volpini, R. A., Camara, N. O., Malheiros, D. M., Shimizu, M. H., Seguro, A. C., & Andrade, L. (2014). Apolipoprotein A-I mimetic peptide 4F attenuates kidney injury, heart injury, and endothelial dysfunction in sepsis. American Journal of Physiology Regulatory, Integrative and Comparative Physiology, 307, R514-524.
Park, S. H., Sung, Y. Y., Jang, S., Nho, K. J., Choi, G. Y., & Kim, H. K. (2016). The Korean herbal medicine, Do In Seung Gi-Tang, attenuates atherosclerosis via AMPK in high-fat diet-induced ApoE(−/−) mice. BMC Complementary and Alternative Medicine, 16, 352.
Yongming, P., Zhaowei, C., Yichao, M., Keyan, Z., Liang, C., Fangming, C., Xiaoping, X., Quanxin, M., & Minli, C. (2015). Involvement of peroxisome proliferator-activated receptors in cardiac and vascular remodeling in a novel minipig model of insulin resistance and atherosclerosis induced by consumption of a high-fat/cholesterol diet. Cardiovascular Diabetology, 14, 6.
Zhao, H., Anand, A. R., & Ganju, R. K. (2014). Slit2-Robo4 pathway modulates lipopolysaccharide-induced endothelial inflammation and its expression is dysregulated during endotoxemia. Journal of Immunology, 192, 385–393.
Mukovozov, I., Huang, Y. W., Zhang, Q., Liu, G. Y., Siu, A., Sokolskyy, Y., Patel, S., Hyduk, S. J., Kutryk, M. J., Cybulsky, M. I., & Robinson, L. A. (2015). The Neurorepellent Slit2 inhibits postadhesion stabilization of monocytes tethered to vascular endothelial cells. The Journal of Immunology, 195, 3334–3344.
Li, C., Yang, G., Lin, L., Xuan, Y., Yan, S., Ji, X., Song, F., Lu, M., & Lan, T. (2019). Slit2 signaling contributes to cholestatic fibrosis in mice by activation of hepatic stellate cells. Experimental Cell Research, 385, 111626.
Yuen, D. A., Huang, Y. W., Liu, G. Y., Patel, S., Fang, F., Zhou, J., Thai, K., Sidiqi, A., Szeto, S. G., Chan, L., Lu, M., He, X., John, R., Gilbert, R. E., Scholey, J. W., & Robinson, L. A. (2016). Recombinant N-terminal Slit2 inhibits TGF-beta-induced fibroblast activation and renal fibrosis. Journal of the American Society of Nephrology, 27, 2609–2615.
Zhou, X., Yao, Q., Sun, X., Gong, X., Yang, Y., Chen, C., & Shan, G. (2017). Slit2 ameliorates renal inflammation and fibrosis after hypoxia-and lipopolysaccharide-induced epithelial cells injury in vitro. Experimental Cell Research, 352, 123–129.
Tajfard, M., Latiff, L. A., Rahimi, H. R., Mouhebati, M., Esmaeily, H., Taghipour, A., Mahdipour, E., Davari, H., Saghiri, Z., Hanachi, P., Ghayour Mobarhan, M., Ferns, G. A., & Azizian, M. (2014). Serum inflammatory cytokines and depression in coronary artery disease. Iranian Red Crescent Medical Journal, 16, e17111.
Kotur-Stevuljevic, J., Spasic, S., Jelic-Ivanovic, Z., Spasojevic-Kalimanovska, V., Stefanovic, A., Vujovic, A., Memon, L., & Kalimanovska-Ostric, D. (2008). PON1 status is influenced by oxidative stress and inflammation in coronary heart disease patients. Clinical Biochemistry, 41, 1067–1073.
Hussain, T., Tan, B., Yin, Y., Blachier, F., Tossou, M. C., & Rahu, N. (2016). Oxidative Stress and Inflammation: What Polyphenols Can Do for Us? Oxidative Medicine and Cellular Longevity, 2016, 7432797.
Miniati, D. N., Lijkwan, M. A., Murata, S., Martens, J., Coleman, C. T., Hoyt, E. G., & Robbins, R. C. (2003). Effects of adenoviral up-regulation of bcl-2 on oxidative stress and graft coronary artery disease in rat heart transplants. Transplantation, 76, 382–386.
Yokota, T., Nomura, K., Nagashima, M., & Kamimura, N. (2016). Fucoidan alleviates high-fat diet-induced dyslipidemia and atherosclerosis in ApoE(shl) mice deficient in apolipoprotein E expression. The Journal of Nutritional Biochemistry, 32, 46–54.
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Liu, JW., Liu, HT. & Chen, L. The Therapeutic Role of Slit2 in Anti-fibrosis, Anti-inflammation and Anti-oxidative Stress in Rats with Coronary Heart Disease. Cardiovasc Toxicol 21, 973–983 (2021). https://doi.org/10.1007/s12012-021-09688-5
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DOI: https://doi.org/10.1007/s12012-021-09688-5