Huang Qian, Sparatore Anna, Del Soldato Piero, Wu Lingyun, Desai Kaushik
Department of Pharmacology, College of Medicine, University of Saskatchewan, Saskatoon, Saskatchewan, Canada; King's Lab, School of Pharmacy, Shanghai Jiao Tong University, Shanghai, China.
Dipartimento di Scienze Farmaceutiche, Università degli Studi di Milano, Milan, Italy.
PLoS One. 2014 Jun 4;9(6):e97315. doi: 10.1371/journal.pone.0097315. eCollection 2014.
Hydrogen sulfide is a gasotransmitter with vasodilatory and anti-inflammatory properties. Aspirin is an irreversible cyclooxygenase inhibitor anti-inflammatory drug. ACS14 is a novel synthetic hydrogen sulfide releasing aspirin which inhibits cyclooxygenase and has antioxidant effects. Methylglyoxal is a chemically active metabolite of glucose and fructose, and a major precursor of advanced glycation end products formation. Methylglyoxal is harmful when produced in excess. Plasma methylglyoxal levels are significantly elevated in diabetic patients. Our aim was to investigate the effects of ACS14 on methylglyoxal levels in cultured rat aortic vascular smooth muscle cells. We used cultured rat aortic vascular smooth muscle cells for the study. Methylglyoxal was measured by HPLC after derivatization, and nitrite+nitrate with an assay kit. Western blotting was used to determine NADPH oxidase 4 (NOX4) and inducible nitric oxide synthase (iNOS) protein expression. Dicholorofluorescein assay was used to measure oxidative stress. ACS14 significantly attenuated elevation of intracellular methylglyoxal levels caused by incubating cultured vascular smooth muscle cells with methylglyoxal (30 µM) and high glucose (25 mM). ACS14, but not aspirin, caused a significant attenuation of increase in nitrite+nitrate levels caused by methylglyoxal or high glucose. ACS14, aspirin, and sodium hydrogen sulfide (NaHS, a hydrogen sulfide donor), all attenuated the increase in oxidative stress caused by methylglyoxal and high glucose in cultured cells. ACS14 prevented the increase in NOX4 expression caused by incubating the cultured VSMCs with MG (30 µM). ACS14, aspirin and NaHS attenuated the increase in iNOS expression caused by high glucose (25 mM). In conclusion, ACS14 has the novel ability to attenuate an increase in methylglyoxal levels which in turn can reduce oxidative stress, decrease the formation of advanced glycation end products and prevent many of the known deleterious effects of elevated methylglyoxal. Thus, ACS14 has the potential to be especially beneficial for diabetic patients pending further in vivo studies.
硫化氢是一种具有血管舒张和抗炎特性的气体递质。阿司匹林是一种不可逆的环氧化酶抑制剂抗炎药物。ACS14是一种新型的释放硫化氢的合成阿司匹林,它能抑制环氧化酶并具有抗氧化作用。甲基乙二醛是葡萄糖和果糖的一种化学活性代谢产物,也是晚期糖基化终末产物形成的主要前体。当甲基乙二醛产生过量时是有害的。糖尿病患者血浆甲基乙二醛水平显著升高。我们的目的是研究ACS14对培养的大鼠主动脉血管平滑肌细胞中甲基乙二醛水平的影响。我们使用培养的大鼠主动脉血管平滑肌细胞进行该研究。甲基乙二醛经衍生化后用高效液相色谱法测定,亚硝酸盐+硝酸盐用试剂盒测定。蛋白质免疫印迹法用于测定NADPH氧化酶4(NOX4)和诱导型一氧化氮合酶(iNOS)蛋白表达。二氯荧光素测定法用于测量氧化应激。用甲基乙二醛(30µM)和高糖(25mM)孵育培养的血管平滑肌细胞所引起的细胞内甲基乙二醛水平升高,ACS14能显著减弱。ACS14而非阿司匹林能显著减弱由甲基乙二醛或高糖引起的亚硝酸盐+硝酸盐水平的升高。ACS14、阿司匹林和硫化氢钠(NaHS,一种硫化氢供体)均能减弱甲基乙二醛和高糖在培养细胞中所引起的氧化应激增加。用MG(30µM)孵育培养的血管平滑肌细胞所引起的NOX4表达增加,ACS14能予以阻止。高糖(25mM)所引起的iNOS表达增加,ACS14、阿司匹林和NaHS均能减弱。总之,ACS14具有减弱甲基乙二醛水平升高的新能力,这反过来又能降低氧化应激,减少晚期糖基化终末产物的形成,并预防甲基乙二醛升高的许多已知有害作用。因此,在进一步的体内研究之前,ACS14可能对糖尿病患者特别有益。