Th17/Treg balance and macrophage polarization ratio in lower extremity arteriosclerosis obliterans

Abstract

Objective: To explore the balance of peripheral blood T helper 17 cells/regulatory T cell (Th17/Treg) ratio and the polarization ratio of M1 and M2 macrophages in lower extremity arteriosclerosis obliterans (ASO). Methods: A rat model of lower extremity ASO was established, and blood samples from patients with lower extremity ASO before and after surgery were obtained. ELISA was used to detect interleukin 6 (IL-6), IL-10, and IL-17. Real-time RCR and Western blot analyses were used to detect Foxp3, IL-6, IL-10, and IL-17 expression. Moreover, flow cytometry was applied to detect the Th17/Treg ratio and M1/M2 ratio. Results: Compared with the control group, the iliac artery wall of ASO rats showed significant hyperplasia, and the concentrations of cholesterol and triglyceride were significantly increased (P<0.01), indicating the successful establishment of ASO. Moreover, the levels of IL-6 and IL-17 in ASO rats were pronouncedly increased (P<0.05), while the IL-10 level was significantly decreased (P<0.05). In addition to increased IL-6 and IL-17 levels, the mRNA and protein levels of Foxp3 and IL-10 in ASO rats were significantly decreased compared with the control group. The Th17/Treg and M1/M2 ratios in the ASO group were markedly increased (P<0.05). These alternations were also observed in ASO patients. After endovascular surgery (such as percutaneous transluminal angioplasty and arterial stenting), all these changes were significantly improved (P<0.05). Conclusions: The Th17/Treg and M1/M2 ratios were significantly increased in ASO, and surgery can effectively improve the balance of Th17/Treg, and reduce the ratio of M1/M2, and the expression of inflammatory factors.

Keywords

Lower extremity arteriosclerosis / Regulatory T cells / Regulatory B cells / Inflammatory factors / M1 macrophages / M2 macrophages

Conflict of interest statement

The authors declare that they have no conflict of interest.

Funding

This work is supported by Natural Science Foundation of Hainan Province (820MS135), Hainan Provincial Health Commission 2023 Provincial Key Clinical Discipline (Clinical Medical Center) Construction Unit Fund Project (Qiongwei Yihan [2022] No. 341), and Hainan Provincial Health Technology Innovation Joint Project (WSJK2024MS209).

Data availability statement

The data supporting the findings of this study are available from the corresponding authors upon request.

Authors’ contributions

CW and YQ participated in study design and data collection. ZD. Liu, ZX, and HQ carried out data analysis. XH drafted the final manuscript. ZZ. Li and ML revised the manuscript. All authors read and approved the final manuscript.

References

[1] Ramanan B, Jeon-Slaughter H, Chen X, Modrall JG, Tsai S. Comparison of open and endovascular procedures in patients with critical limb ischemia on dialysis. J Vasc Surg 2019; 70(4): 1217-1224.

[2] Libby P. Inflammation during the life cycle of the atherosclerotic plaque. Cardiovasc Res 2021; 117(13): 2525-2536.

[3] Babaniamansour P, Mohammadi M, Babaniamansour S, Aliniagerdroudbari E. The relation between atherosclerosis plaque composition and plaque rupture. J Med Signals Sens 2020; 10(4): 267-273.

[4] Chen B, Huang S, Su Y, Wu YJ, Hanna A, Brickshawana A, et al. Macrophage Smad3 protects the infarcted heart, stimulating phagocytosis and regulating inflammation. Circ Res 2019; 125(1): 55-70.

[5] Chen X, Tang J, Shuai W, Meng J, Feng J, Han Z. Macrophage polarization and its role in the pathogenesis of acute lung injury/acute respiratory distress syndrome. Inflamm Res 2020; 69(9): 883-895.

[6] Gorgui J, Gasbarrino K, Georgakis MK, Karalexi MA, Nauche B, Petridou ET, et al. Circulating adiponectin levels in relation to carotid atherosclerotic plaque presence, ischemic stroke risk, and mortality: A systematic review and meta-analyses. Metabolism 2017; 69: 51-66.

[7] Arce-Sillas A, Álvarez-Luquín DD, Tamaya-Domínguez B, Gomez-Fuentes S, Trejo-García A, Melo-Salas M, et al. Regulatory T cells: Molecular actions on effector cells in immune regulation. J Immunol Res 2016; 2016. doi: 10.1155/2016/1720827.

[8] Tabas I, Bornfeldt KE. Macrophage phenotype and function in different stages of atherosclerosis. Circ Res 2016; 118(4): 653-667.

[9] Gisterå A, Hansson GK. The immunology of atherosclerosis. Nat Rev Nephrol 2017; 13(6): 368-380.

[10] Liu Y, Wang CQ, Zhu YK, Xu JF, Yin SQ, Hu QJ, et al. Macrophage-secreted exosomes inhibit breast cancer cell migration via the miR-101-3p/DLG5 axis. Asian Pac J Trop Biomed 2023; 13(12): 532-538.

[11] Taghizadeh E, Taheri F, Renani PG, Reiner Ž, Navashenaq JG, Sahebkar A. Macrophage: A key therapeutic target in atherosclerosis? Curr Pharm Des 2019; 25(29): 3165-3174.

[12] Wolf D, Ley K. Immunity and inflammation in atherosclerosis. Circ Res 2019; 124(2): 315-327.

[13] Kasahara K, Sasaki N, Amin HZ, Tanaka T, Horibe S, Yamashita T, et al. Depletion of Foxp3+ regulatory T cells augments CD4+ T cell immune responses in atherosclerosis-prone hypercholesterolemic mice. Heliyon 2022; 8(7): e09981.

[14] Huang Y, Hu H, Liu L, Ye J, Wang Z, Que B, et al. Interleukin-12p35 deficiency reverses the Th1/Th2 imbalance, aggravates the Th17/Treg imbalance, and ameliorates atherosclerosis in ApoE ^-/- mice . Mediators Inflamm 2019; 2019. doi: 10.1155/2019/3152040.

[15] Zhang W, Liu X, Zhu Y, Liu X, Gu Y, Dai X, et al. Transcriptional and posttranslational regulation of Th17/Treg balance in health and disease. Eur J Immunol 2021; 51(9): 2137-2150.

[16] Tulis DA. Histological and morphometric analyses for rat carotid balloon injury model. Methods Mol Med 2007; 139: 31-66.

[17] Li Q, Kuang Y, Qiu J, Zhang X, Ruan Y, Li Z. The correlation between plasma tissue factor and interleukin 18 and their significance in patients with acute coronary syndrome. Cardiovasc Toxicol 2015; 15(3): 276-282.

[18] Schuett H, Luchtefeld M, Grothusen C, Grote K, Schieffer B. How much is too much? Interleukin-6 and its signalling in atherosclerosis. Thromb Haemost 2009; 102(2): 215-222.

[19] Tyrrell DJ, Goldstein DR. Ageing and atherosclerosis: Vascular intrinsic and extrinsic factors and potential role of IL-6. Nat Rev Cardiol 2021; 18(1): 58-68.

[20] Montgomery A, Tam F, Gursche C, Cheneval C, Besler K, Enns W, et al. Overlapping and distinct biological effects of IL-6 classic and trans-signaling in vascular endothelial cells. Am J Physiol Cell Physiol 2021; 320(4): C554-C565.

[21] O’Garra A, Barrat FJ, Castro AG, Vicari A, Hawrylowicz C. Strategies for use of IL-10 or its antagonists in human disease. Immunol Rev 2008; 223: 114-131.

[22] Saraiva M, Vieira P, O’Garra A. Biology and therapeutic potential of interleukin-10. J Exp Med 2020; 217(1). doi: 10.1084/jem.20190418.

[23] Pinderski Oslund LJ, Hedrick CC, Olvera T, Hagenbaugh A, Territo M, Berliner JA, et al. Interleukin-10 blocks atherosclerotic events in vitro and in vivo. Arterioscler Thromb Vasc Biol 1999; 19(12): 2847-2853.

[24] Zhang DF, Song XT, Chen YD, Yuan F, Xu F, Zhang M, et al. Prognostic performance of interleukin-10 in patients with chest pain and mild to moderate coronary artery lesions-an 8-year follow-up study. J Geriatr Cardiol 2016; 13(3): 244-251.

[25] Heeschen C, Dimmeler S, Hamm CW, Fichtlscherer S, Boersma E, Simoons ML, et al. Serum level of the antiinflammatory cytokine interleukin-10 is an important prognostic determinant in patients with acute coronary syndromes. Circulation 2003; 107(16): 2109-2114.

[26] McGeachy MJ, Cua DJ, Gaffen SL. The IL-17 Family of cytokines in health and disease. Immunity 2019; 50(4): 892-906.

[27] Gong F, Liu Z, Liu J, Zhou P, Liu Y, Lu X. The paradoxical role of IL-17 in atherosclerosis. Cell Immunol 2015; 297(1): 33-39.

[28] Novikova OA, Laktionov PP, Karpenko AA. The roles of mechanotransduction, vascular wall cells, and blood cells in atheroma induction. Vascular 2019; 27(1): 98-109.

[29] Nilsson J, Hansson GK. Vaccination strategies and immune modulation of atherosclerosis. Circ Res 2020; 126(9): 1281-1296.

[30] Li M, Yang C, Wang Y, Song W, Jia L, Peng X, et al. The expression of P2X7 receptor on Th1, Th17, and regulatory T cells in patients with systemic lupus erythematosus or rheumatoid arthritis and its correlations with active disease. J Immunol 2020; 205(7): 1752-1762.

[31] Joetham A, Schedel M, Ning F, Wang M, Takeda K, Gelfand EW. Dichotomous role of TGF-β controls inducible regulatory T-cell fate in allergic airway disease through Smad3 and TGF-β-activated kinase 1. J Allergy Clin Immunol 2020; 145(3): 933-946.e4.

[32] Bhaumik S, Mickael ME, Moran M, Spell M, Basu R. RORγt promotes Foxp3 expression by antagonizing the effector program in colonic regulatory T cells. J Immunol 2021; 207(8): 2027-2038.

[33] Thota LN, Ponnusamy T, Lu X, Mundkur L. Long-Term efficacy and safety of immunomodulatory therapy for atherosclerosis. Cardiovasc Drugs Ther 2019; 33(4): 385-398.

[34] Batlle E, Massagué J. Transforming growth factor-β signaling in immunity and cancer. Immunity 2019; 50(4): 924-940.

[35] Libby P, Pasterkamp G, Crea F, Jang IK. Reassessing the mechanisms of acute coronary syndromes. Circ Res 2019; 124(1): 150-160.

[36] Gaba P, Gersh BJ, Muller J, Narula J, Stone GW. Evolving concepts of the vulnerable atherosclerotic plaque and the vulnerable patient: Implications for patient care and future research. Nat Rev Cardiol 2023; 20(3): 181-196.