羟基红花黄色素A对氧化应激损伤血管内皮细胞的保护作用研究
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摘要
目的探讨羟基红花黄色素A(hydroxysafflor yellow A, HSYA)对血管内皮细胞氧化应激损伤的保护作用及其可能机制。方法体外培养人血管内皮细胞EC-304,用H_2O_2构建细胞氧化应激损伤模型,分别设置正常对照组、H_2O_2损伤模型组、HSYA药物组,其中各药物组用不同浓度的HSYA预培养细胞24 h后加入50μmol/L的H_2O_2,继续培养12 h。用MTT法检测细胞的增殖活力,用试剂盒检测各组细胞内超氧化物歧化酶(SOD)、一氧化氮(NO)的含量,用Western Blot法检测各组细胞Bax、Bcl-2、Caspase-3、cleaved Caspase-3蛋白表达水平。结果与H_2O_2模型组相比,HSYA能显著提高细胞存活率,且呈剂量依赖性,提高细胞内SOD的活性,提高细胞内NO的含量,降低Bax表达,提高Bcl-2表达,降低Caspase-3和cleaved Caspase-3的表达(P<0.01或P<0.05)。结论 HSYA对于H_2O_2诱导的血管内皮细胞氧化应激损伤具有保护作用,其可能的作用机制与抑制EC-304细胞凋亡相关。
Objective To study the protective mechanism of hydroxysafflor yellow A(HSYA) on vascular endothelial cells injured by oxidative stress. Methods EC-304 human vascular endothelial cells were cultured in vitro. An oxidative stress injury model was established with H_2O_2. The cells were divided into several groups, namely control group, H_2O_2 injury model group,and HSYA groups. The cells in HSYA groups were pre-cultured with different concentrations of HSYA for 24 h, followed by addition of 50 μmol/L H_2O_2, and the cells were then cultured for another 12 h. Cell proliferation activity was determined by MTT assay, the content of superoxide dismutase(SOD) and nitric oxide(NO) was measured by kits, and the protein expression levels of Bax, Bcl-2, Caspase-3, and cleaved Caspase-3 were determined by Western Blot. Results Compared with the H_2O_2 model group,HSYA significantly increased the cell survival rate in a dose-dependent manner, and it significantly increased SOD activity, NO content, and Bcl-2 expression and significantly reduced the expression of Bax, Caspase-3, and cleaved Caspase-3(P<0.01 or P<0.05). Conclusion HSYA has a protective effect on H_2O_2-induced oxidative stress injury of vascular endothelial cells, and the mechanism is possibly related to inhibition of the apoptosis of EC-304 cells.
Objective To study the protective mechanism of hydroxysafflor yellow A(HSYA) on vascular endothelial cells injured by oxidative stress. Methods EC-304 human vascular endothelial cells were cultured in vitro. An oxidative stress injury model was established with H_2O_2. The cells were divided into several groups, namely control group, H_2O_2 injury model group,and HSYA groups. The cells in HSYA groups were pre-cultured with different concentrations of HSYA for 24 h, followed by addition of 50 μmol/L H_2O_2, and the cells were then cultured for another 12 h. Cell proliferation activity was determined by MTT assay, the content of superoxide dismutase(SOD) and nitric oxide(NO) was measured by kits, and the protein expression levels of Bax, Bcl-2, Caspase-3, and cleaved Caspase-3 were determined by Western Blot. Results Compared with the H_2O_2 model group,HSYA significantly increased the cell survival rate in a dose-dependent manner, and it significantly increased SOD activity, NO content, and Bcl-2 expression and significantly reduced the expression of Bax, Caspase-3, and cleaved Caspase-3(P<0.01 or P<0.05). Conclusion HSYA has a protective effect on H_2O_2-induced oxidative stress injury of vascular endothelial cells, and the mechanism is possibly related to inhibition of the apoptosis of EC-304 cells.
引文
[1] CHEN X, PANG S, LIN J, et al. Allicin prevents oxidized low-density lipoprotein-induced endothelial cell injury by inhibiting apoptosis and oxidative stress pathway[J]. BMC Complementary&Alternative Medicine, 2016,16(1):1-6.
[2]陈梦,赵丕文,孙丽萍,等.羟基红花色素A对雌激素效应相关蛋白表达的影响[J].湖南中医药大学学报,2017,37(11):1218-1221.
[3] SUI X, DOU L, CHEN X, et al. miR-291b-3p mediated ROSinduced endothelial cell dysfunction by targeting HUR[J]. International Journal of Molecular Medicine, 2018, 42(5):2383-2392.
[4] NIKE E. Oxidant-specific biomarkers of oxidative stress. Association with atherosclerosis and implication for antioxidant effects[J].Free Radical Biology And Midicine, 2018,120:425-440.
[5] STEINHORN B S, LOSCALZO J, MICHEL T. Nitroglycerin and Nitric Oxide-A Rondo of Themes in Cardiovascular Therapeutics[J].New England Journal of Medicine, 2015, 373(18):277-280.
[6] YU Y, ZHANG X, LI Z, et al. LncRNA HOTAIR suppresses TNF-αinduced apotosis of nucleus pulposus cells by regulating miR-34a/Bcl-2 axis[J]. Biomedicine&Pharmacotherapy, 2018,107:729-737.
[7] OSMAN A M, NEUMANN S, KUHN H G, et al. Caspase inhibition impaired the neural stem/progenitor cell response after cortical ischemia in mice[J]. Oncotarget, 2016, 7(3):2239-2248.
[8] KURIYAMA S, TSUJI T, SAKUMA T, et al. PLEKHN1 promotes apoptosis by enhancing Bax-Bak hetro-oligomerization through interaction with Bid in human colon cancer[J]. Cell Death Discovery, 2018,4:11.
[9] LOPEZ J, TAIT S W. Mitochondrial apoptosis:killing cancer using the enemy within[J]. British Journal Cancer, 2015,112(6):957-962.
[10] JUNG S Y, KIM D Y, YUNE T Y, et al. Treadmill exercise reduces spinal cord injury-induced apoptosis by activating the P13K/Akt pathway in rats[J]. Experimental&Therapeutic Medicine,2014,7(3):587-593.
[11] DING L, WU J P, XU G, et al. Lentiviral-mediated RNAi targeting caspase-3 inhibits apoptosis induced by serum deprivation in rat endplate chondrocytes in vitro[J]. Brazilian Journal of Medical&Biological Research, 2014,47(6):445-451.
[12] ZHAO Y, SUN H, LI X, et al. Hydroxysafflor yellow A attenuates high glucose-induced pancreaticβ-cells oxidative damage via inhibiting JNK/c-jun signaling pathway[J]. Biochemical and Biophysical Research Communications, 2018,505(2):425-440.
[13] PEI J P, FAN L H, NAN K, et al. HSYA alleviates secondary neuronal death through attenuating oxidative stress, inflammatory response, and neural apoptosis in SD rat spinal cord compression injury[J]. Journal of Neuroinflammation, 2017,14(1):97.
[2]陈梦,赵丕文,孙丽萍,等.羟基红花色素A对雌激素效应相关蛋白表达的影响[J].湖南中医药大学学报,2017,37(11):1218-1221.
[3] SUI X, DOU L, CHEN X, et al. miR-291b-3p mediated ROSinduced endothelial cell dysfunction by targeting HUR[J]. International Journal of Molecular Medicine, 2018, 42(5):2383-2392.
[4] NIKE E. Oxidant-specific biomarkers of oxidative stress. Association with atherosclerosis and implication for antioxidant effects[J].Free Radical Biology And Midicine, 2018,120:425-440.
[5] STEINHORN B S, LOSCALZO J, MICHEL T. Nitroglycerin and Nitric Oxide-A Rondo of Themes in Cardiovascular Therapeutics[J].New England Journal of Medicine, 2015, 373(18):277-280.
[6] YU Y, ZHANG X, LI Z, et al. LncRNA HOTAIR suppresses TNF-αinduced apotosis of nucleus pulposus cells by regulating miR-34a/Bcl-2 axis[J]. Biomedicine&Pharmacotherapy, 2018,107:729-737.
[7] OSMAN A M, NEUMANN S, KUHN H G, et al. Caspase inhibition impaired the neural stem/progenitor cell response after cortical ischemia in mice[J]. Oncotarget, 2016, 7(3):2239-2248.
[8] KURIYAMA S, TSUJI T, SAKUMA T, et al. PLEKHN1 promotes apoptosis by enhancing Bax-Bak hetro-oligomerization through interaction with Bid in human colon cancer[J]. Cell Death Discovery, 2018,4:11.
[9] LOPEZ J, TAIT S W. Mitochondrial apoptosis:killing cancer using the enemy within[J]. British Journal Cancer, 2015,112(6):957-962.
[10] JUNG S Y, KIM D Y, YUNE T Y, et al. Treadmill exercise reduces spinal cord injury-induced apoptosis by activating the P13K/Akt pathway in rats[J]. Experimental&Therapeutic Medicine,2014,7(3):587-593.
[11] DING L, WU J P, XU G, et al. Lentiviral-mediated RNAi targeting caspase-3 inhibits apoptosis induced by serum deprivation in rat endplate chondrocytes in vitro[J]. Brazilian Journal of Medical&Biological Research, 2014,47(6):445-451.
[12] ZHAO Y, SUN H, LI X, et al. Hydroxysafflor yellow A attenuates high glucose-induced pancreaticβ-cells oxidative damage via inhibiting JNK/c-jun signaling pathway[J]. Biochemical and Biophysical Research Communications, 2018,505(2):425-440.
[13] PEI J P, FAN L H, NAN K, et al. HSYA alleviates secondary neuronal death through attenuating oxidative stress, inflammatory response, and neural apoptosis in SD rat spinal cord compression injury[J]. Journal of Neuroinflammation, 2017,14(1):97.