Expression of 3-nitrotyrosine, iNOS and NO in brain tissues of ischemia-reperfusion rats treated with different doses of hydroxysafflor yellow A

XIAO Chen; SUN Li; CAO Shanshan; XU Yanwei; LIANG Hao; CHENG Yan

doi:10.3969/j.issn.1002-266X.2017.11.005

Abstract：Objective To observe the expression changes of 3-nitrotyrosine (3-NT), inducible nitric oxide synthase (iNOS), and nitric oxide (NO) in brain tissues of ischemia-reperfusion rats treated with different doses of hydroxysafflor yellow A (HSYA) by intravenous injection.Methods The healthy adult male SD rats were randomly divided into the sham operation group, model group, low-dose HSYA group, medium-dose HSYA group and high-dose HSYA group.In the sham operation group, rats' common carotid artery, internal carotid artery and external carotid artery were exposed, and then sutured.The other rats were subjected to a 60-min middle cerebral artery occlusion and 24-h reperfusion (MCAO/R).The rats in the low-dose, medium-dose and high-dose HSYA groups were injected with 2.5, 5 and 10 mg/kg HSYA, respectively, in the tail vein at 60 min after ischemia, and the same volume of Tris buffer was injected into the tail vein of rats in sham operation group and model group.All rats were continued to raise for 24 h.The expression of 3-NT and iNOS in the rat brain tissues after ischemia-reperfusion in each group was examined by Western blotting.The NO level was also detected by Griess assay.Results The absolute gray values of 3-NT in the sham operation group, the model group, the low-dose HSYA group, the medium-dose HSYA group and the high-dose HSYA group after ischemia-reperfusion were (2.81±1.37)×103, (46.86±4.75)×103, (44.51±4.13)×103, (13.88±2.98)×103, and (6.38±2.66)×103, respectively;the absolute gray values of iNOS of each group were (2.25±0.32)×103, (79.67±4.73)×103, (75.29±4.08)×103, (27.31±2.77)×103, (19.19±1.86)×103, respectively.In the model group and low-dose, medium-dose, and high-dose HSYA groups, the levels of NO were (16.50±2.20), (15.40±1.44), (10.33±1.30), and (6.80±0.73) nmol/mg.The levels of 3-NT, iNOS and NO in brain tissues of the model group, low-dose HSYA group, medium-dose HSYA group and high-dose HSYA group were higher than those of the sham operation group (all P<0.05).The levels of 3-NT, iNOS and NO in the brain tissues of rats in the low-dose HSYA group were not significantly different from those in the model group (P>0.05).The levels of 3-NT, iNOS, NO in the medium-dose and high-dose HSYA groups were lower than those in the model group and low-dose HSYA group (all P<0.05), and the levels of 3-NT, iNOS and NO in brain tissues of high-dose HSYA group were lower than those in medium-dose HSYA group (all P<0.05).Conclusion HSYA can inhibit the expression of 3-NT, iNOS, and NO in rat brain tissues after ischemia-reperfusion.The higher the HSYA dose is, the more obvious the effect will be.