Zehra ÇİÇEK, Kübra AKILLIOĞLU, Ayşe DOĞAN

The Effects of Angiotensin II, Olmesartan and PD123,319 on Protein Kinase C in STZ Induced Rat Vascular Smooth Muscle Cell Culture

Protein kinase C (PKC) is a promoter enzyme that plays a vital role in signal transduction of vascular smooth muscle cells (VSMCs). It has numerous vascular functions, such as vascular cell growth, cytokine activation and angiogenesis. But, these mechanisms are deteriorating in diabetes mellitus. Angiotensin II (Ang II) effects on vascular structure binding Ang II type-1 and type-2 receptors (ATR1 and ATR2) and stimulates PKC mostly through ATR1s. We aim to investigate the effects of Ang II, Olmesartan and PD123,319 on PKC levels of healthy and streptozotocin (STZ) induced rat VSMCs (H-VSMCs and STZ-VSMCs) in this study. The primary culture of VSMCs were isolated from thoracic aorta of healthy and STZ (45 mg/kg, given via the tail vein) induced Wistar rats. PKC levels were measured in H-VSMCs and STZ-VSMCs by enzyme-linked immunosorbent assay (ELISA). In H-VSMCs, Ang II group compared to the control group, PKC levels decreased significantly (P = 0.000); whereas, Ang II group compared to control, PKC levels were higher, but not significantly in STZ-VSMCs (P = 0.088). PKC levels were increased in Ang II+Olmesartan (P = 0.000) and Ang II+PD123,319 (P = 0.000) groups compared to Ang II group in H-VSMCs, but in STZ-VSMCs, PKC levels in Ang II+Olmesartan (P = 0.001) and Ang II+PD123,319 (P = 0.000) groups compared to Ang II group were decreased significantly. Ang II, its receptors and PKC seem to modulate each other and may have a relationship in hyperglycemic conditions. Also, considering that, ATR1 blocker (ATR1B) Olmesartan and ATR2 blocker (ATR2B) PD123,319 may be protective against vascular injury by reducing PKC levels in STZ-VSMCs.

Anahtar Kelimeler:

Ang II, PKC, Vascular smooth muscle, Cell culture

The Effects of Angiotensin II, Olmesartan and PD123,319 on Protein Kinase C in STZ Induced Rat Vascular Smooth Muscle Cell Culture

Keywords:

Ang II, PKC, Vascular smooth muscle, Cell culture,

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Agata J, Ura N, Yoshida H, Shinshi Y, Sasaki H, Hyakkoku M, Shimamoto K. 2006. Olmesartan is an angiotensin II receptor blocker with an inhibitory effect on angiotensin-converting enzyme. Hypertension Res, 29(11): 865-874. doi: DOI 10.1291/hypres.29.865.
American Diabetes A. 2009. Diagnosis and classification of diabetes mellitus. Diabetes Care, 32(Suppl 1): 62-67. doi: 10.2337/dc09-S062.
Chamley-Campbell J, Campbell G. R, Ross R. 1979. The smooth muscle cell in culture. Physiological Rev, 59(1): 1-61. doi: 10.1152/physrev.1979.59.1.1.
Chi J, Meng L, Pan S, Lin H, Zhai X, Liu L, . . . Guo H. 2017. Primary Culture of Rat Aortic Vascular Smooth Muscle Cells: A New Method. Med Sci Monit, 23: 4014-4020. doi: 10.12659/msm.902816.
Daryabor G, Atashzar M. R, Kabelitz D, Meri S, Kalantar K. 2020. The effects of type 2 diabetes mellitus on organ metabolism and the immune system. Front Immunol, 11: 1582. doi: 10.3389/fimmu.2020.01582.
Das Evcimen N, King G. L. 2007. The role of protein kinase C activation and the vascular complications of diabetes. Pharmacol Res, 55(6): 498-510. doi: 10.1016/j.phrs.2007.04.016.
Forrester SJ, Booz GW, Sigmund CD, Coffman TM, Kawai T, Rizzo V, . . . Eguchi S. 2018. Angiotensin II Signal Transduction: An Update on Mechanisms of Physiology and Pathophysiology. Physiol Rev, 98(3): 1627-1738. doi: 10.1152/physrev.00038.2017.
Geraldes P, King G. L. 2010. Activation of protein kinase C isoforms and its impact on diabetic complications. Circ Res 106(8): 1319-1331. doi: 10.1161/CIRCRESAHA.110.217117.
Hinz B, Celetta G, Tomasek J. J, Gabbiani G, Chaponnier C. 2001. Alpha-smooth muscle actin expression upregulates fibroblast contractile activity. Mol Biol Cell, 12(9): 2730-2741. doi: 10.1091/mbc.12.9.2730.
Jaminon A, Reesink K, Kroon A, Schurgers L. 2019. The role of vascular smooth muscle cells in arterial remodeling: focus on calcification-related processes. Int J Mol Sci, 20(22): 5694. doi: 10.3390/ijms20225694.
Kizub IV, Klymenko KI, Soloviev AI. 2014. Protein kinase C in enhanced vascular tone in diabetes mellitus. Int J Cardiol, 174(2): 230-242. doi: 10.1016/j.ijcard.2014.04.117.
Lien CF, Chen SJ, Tsai MC, Lin CS. 2021. Potential role of protein kinase c in the pathophysiology of diabetes-associated atherosclerosis. Front Pharmacol, 12: 716332. doi: 10.3389/fphar.2021.716332.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. 1951. Protein measurement with the Folin phenol reagent. J Biol Chem, 193(1): 265-275.
Malhotra A, Kang BP, Cheung S, Opawumi D, Meggs LG. 2001. Angiotensin II promotes glucose-induced activation of cardiac protein kinase C isozymes and phosphorylation of troponin I. Diabetes, 50(8): 1918-1926. doi: 10.2337/diabetes.50.8.1918.
Malhotra A, Reich D, Reich D, Nakouzi A, Sanghi V, Geenen DL, Buttrick PM. 1997. Experimental diabetes is associated with functional activation of protein kinase C epsilon and phosphorylation of troponin I in the heart which are prevented by angiotensin II receptor blockade. Circ Res, 81(6): 1027-1033. doi: 10.1161/01.res.81.6.1027.
Mehta PK, Griendling KK. 2007. Angiotensin II cell signaling: physiological and pathological effects in the cardiovascular system. Am J Physiol Cell Physiol, 292(1): 82-97. doi: 10.1152/ajpcell.00287.2006.
Naudi A, Jove M, Ayala V, Cassanye A, Serrano J, Gonzalo H, . . . Pamplona R. 2012. Cellular dysfunction in diabetes as maladaptive response to mitochondrial oxidative stress. Exp Diabetes Res, 2012:696215. doi: 10.1155/2012/696215.
Pacurari M, Kafoury R, Tchounwou PB, Ndebele K. 2014. The Renin-Angiotensin-aldosterone system in vascular inflammation and remodeling. Int J Inflam, 2014: 689360. doi: 10.1155/2014/689360.
Paz Ocaranza M, Riquelme JA, Garcia L, Jalil JE, Chiong M, Santos RAS, Lavandero S. 2020. Counter-regulatory renin-angiotensin system in cardiovascular disease. Nat Rev Cardiol, 17(2): 116-129. doi: 10.1038/s41569-019-0244-8.
Rameshrad M, Babaei H, Azarmi Y, Fouladia D. F. 2016. Rat aorta as a pharmacological tool for in vitro and in vivo studies. Life Sciences 145 190-204. doi: 10.1016/j.lfs.2015.12.043.
Rask-Madsen C, King G. L. 2013. Vascular complications of diabetes: mechanisms of injury and protective factors. Cell Metab, 17(1): 20-33. doi: 10.1016/j.cmet.2012.11.012.
Sharma V, Sharma PL. 2013. Role of different molecular pathways in the development of diabetes-induced nephropathy. J Diabetes Metabol, 4: 1-7. doi: 10.4172/2156-6156.S9-004.
Steinberg SF. 2008. Structural basis of protein kinase C isoform function. Physiological Rev, 88(4): 1341-1378. doi: 10.1152/physrev.00034.2007.
Touyz R. M, Schiffrin E. L. 2000. Signal transduction mechanisms mediating the physiological and pathophysiological actions of angiotensin II in vascular smooth muscle cells. Pharmacol Rev 52(4): 639-672.
Waterborg JH, Matthews HR. 1994. The Lowry method for protein quantitation. Quantum Dots 3 Edition, 32: 1-4. doi: 10.1385/0-89603-268-X:1.