The Inhibition Effect And Mechanisms Of Resveratrol On Vascular Calcification Induced By Vitamin D3 Plus Nicotine In Rats

Background:Vascular calcification (VC), a common complication and clinical feature of atherosclerosis, diabetes mellitus, chronic kidney disease (CKD) and aging, is believed to be an important risk factor of cardiovascular morbidity and mortality and considered a strong prognostic marker. It is characterized by decreased arterial wall elasticity and impeded blood flow and can result in heart attacks and stroke. Growing evidence suggests that VC, similar to bone remodeling, is an actively regulated, preventable and reversible cell-mediated process, including both inductive and inhibitory processes.Resveratrol (RSV) is a naturally occurring antioxidant polyphenol found in plants such as grapes, mulberries, eucalyptus and peanuts. Accumulating evidence shows that resveratrol protects the cardiovascular system by a large number of mechanisms including improving endothelial function, decreasing myocardial ischemic reperfusion injury, promotion of vasorelaxation, anti-inflammatory, anti-oxidative, anti-proliferative, anti-atherosclerotic properties, inhibition of modification of low-density lipoproteins, estrogen-like actions, anti-platelet aggregation and anti-thrombotic properties.Objective:To investigate the effects of resveratrol on hemodynamic parameters in rats with VC; To investigate the effects of resveratrol on ALP activity in serum or aorta, serum ionized calcium level, aortic calcium content and calcium deposition; To investigate the effects of resveratrol on mRNA and protein of OPN, BMP-2 and Cbfa-1 in aorta of rats with VC.Methods:32 Male Sprague-Dawley rats were randomly assigned to 4 groups (n=8):normal control group (Con), given the same dose of saline and peanut oil; calcification group (VDN), given vitamin D3 (300,000U/kg, intramuscularly) plus nicotine (25mg/kg in 5ml peanut oil, intragastrically); VDN+RSV[L] group, VDN plus resveratrol (25mg/kg/day, intragastrically); VDN+RSV[H] group, VDN plus resveratrol (50mg/kg/day, intragastrically). Hemodynamic parameters were measured by use of the Powerlab Biological System; ALP activity in serum or aorta, serum ionized calcium level and aortic calcium content were measured by use of detection kit; Aortic structure and calcium deposition were detected by use of HE and Von Kossa staining. The expression of OPN, BMP-2 and Cbfα-1 were identified by use of RT-PCR and Western blot.Results:1. Compared with Con group, LVW/BW, HR, SBP, PP, MBP and LVSP in VDN group were increased respectively (all P< 0.05). Compared with VDN group, PP in VDN+RSV[L] group and LVW/BW, SBP, PP, MBP and LVSP in VDN+RSV[H] group were decreased respectively (all P< 0.05). Compared with VDN+RSV[L] group, SBP, PP and LVSP in VDN+RSV [H] group were decreased respectively (all P< 0.05).2. Compared with Con group, HE and von kossa staining revealed thickened vessel walls, disordered elastic fibers and widespread dark silver-staining granules in VDN group; However, the thickness of the aortic, irregular elastic fibers wall and dark silver-staining granules in VDN+RSV[L] and VDN+RSV[H] groups were reduced respectively.3. Compared with Con group, ALP activity in serum or aorta and calcium content in aorta in VDN group were increased respectively (all P< 0.05); Compared with VDN group, ALP activity in serum or aorta and calcium content in aorta in VDN+RSV[L] and VDN+RSV[H] groups were decreased respectively (all P< 0.05); Compared with VDN+RSV[L] group, ALP activity in serum or aorta and calcium content in aorta in VDN+RSV[H] group were decreased respectively (all P< 0.05).4. Compared with Con group, the mRNA and protein of OPN, BMP-2 and Cbfa-1 in VDN group were decreased respectively (all P< 0.05). Compared with VDN group, the mRNA and protein of OPN, BMP-2 and Cbfα-1 in VDN+RSV[L] and VDN+RSV[H] groups were decreased respectively (all P<0.05). Compared with VDN+RSV[L] group, the mRNA of BMP-2 and the protein of OPN in VDN+RSV[H] group were decreased respectively (all P< 0.05).Conclusion:1. Resveratrol reduced blood pressure and improved cardiac function in rats with VC; 2. Resveratrol inhibits VC induced by vitamin D3 plus nicotine in rats.