Urantide对动脉粥样硬化大鼠肝细胞外调节蛋白激酶1/2的作用及机制
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摘要
目的:观察Urantide对动脉粥样硬化(AS)大鼠肝中细胞外调节蛋白激酶1/2(ERK1/2)表达的影响,探讨AS大鼠脂肪肝中调控ERK1/2的分子机制。方法:维生素D3(VD3)腹腔注射及特制高脂饲料饮食建立大鼠AS模型,随机分为正常组、AS组、辛伐他汀组、Urantide组。H-E染色观察大鼠胸主动脉、肝的形态学改变;生化检测大鼠血清中总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白(HDL)、低密度脂蛋白(LDL)的含量;免疫印迹法检测各组大鼠肝组织中尾加压素Ⅱ(UⅡ)、G蛋白偶联受体14(GPR14)、磷酸化ERK1/2(p-ERK1/2)、ERK1/2蛋白质表达水平;免疫荧光法检测各组大鼠肝细胞中p-ERK1/2的表达水平。结果:与正常组相比,AS组大鼠胸主动脉出现典型的AS病理学改变,肝出现典型的脂肪变性;AS组大鼠血清中TC、TG、LDL含量显著升高,HDL含量显著降低;大鼠肝中UⅡ、GPR14、p-ERK1/2、ERK1/2的蛋白表达水平显著升高。与AS组相比,辛伐他汀组及Urantide组大鼠肝的脂肪变性明显减轻;肝中UⅡ、GPR14、p-ERK1/2蛋白表达显著降低,ERK1/2蛋白表达水平无显著变化。结论:Urantide可通过抑制UⅡ/GPR14的生物学效应抑制ERK1/2的活化进而达到治疗脂肪肝的作用。
Objective:To observe the effect of Urantide on the expression of ERK1/2 in the liver of atherosclerotic(AS) rats and investigate the molecular mechanisms contributing to ERK1/2 in fatty liver of AS rats. Methods :The rat AS model was established by intraperitoneal injection of vitamin D3(VD3) into rats and feeding with special high fat food.According to the principle of randomization,the rats were divided into control group,AS group,Simvastatin group and Urantide group. H-E staining was used to observe the morphological changes of the thoracic aorta and liver of the rats. Biochemical indexes were measured to detect the change of total cholesterol(TC),trilaurin(TG),high density lipoprotein(HDL),and low density lipoprotein(LDL) in the serum of rats. The expression levels of urotensin Ⅱ(UⅡ),GPR14,p-ERK1/2 and ERK1/2 protein in liver tissues were detected by Western blotting. Immunofluorescence was used to detect the expression level of p-ERK1/2 in hepatocytes of each group. Results :Compared with control group,the H-E staining of AS group showed typical AS pathological changes and steatosis of the liver;the levels of TC,TG and LDL in AS group were significantly increased,and the level of HDL was significantly decreased;the protein levels of UⅡ,GPR14,p-ERK1/2,ERK1/2 in the liver were significantly increased in AS group. Compared with AS group,steatosis of the liver in Simvastatin group and Urantide group was significantly alleviated;the protein levels of UⅡ,GPR14,p-ERK1/2 in the liver were significantly decreased and no change was observed in the protein levels of ERK1/2 in Simvastatin group and Urantide group. Conclusion :Urantide can inhibit the activation of ERK1/2 by inhibiting the biological effects of UⅡ/GPR14,thus achieving the therapeutic effect on fatty liver.
Objective:To observe the effect of Urantide on the expression of ERK1/2 in the liver of atherosclerotic(AS) rats and investigate the molecular mechanisms contributing to ERK1/2 in fatty liver of AS rats. Methods :The rat AS model was established by intraperitoneal injection of vitamin D3(VD3) into rats and feeding with special high fat food.According to the principle of randomization,the rats were divided into control group,AS group,Simvastatin group and Urantide group. H-E staining was used to observe the morphological changes of the thoracic aorta and liver of the rats. Biochemical indexes were measured to detect the change of total cholesterol(TC),trilaurin(TG),high density lipoprotein(HDL),and low density lipoprotein(LDL) in the serum of rats. The expression levels of urotensin Ⅱ(UⅡ),GPR14,p-ERK1/2 and ERK1/2 protein in liver tissues were detected by Western blotting. Immunofluorescence was used to detect the expression level of p-ERK1/2 in hepatocytes of each group. Results :Compared with control group,the H-E staining of AS group showed typical AS pathological changes and steatosis of the liver;the levels of TC,TG and LDL in AS group were significantly increased,and the level of HDL was significantly decreased;the protein levels of UⅡ,GPR14,p-ERK1/2,ERK1/2 in the liver were significantly increased in AS group. Compared with AS group,steatosis of the liver in Simvastatin group and Urantide group was significantly alleviated;the protein levels of UⅡ,GPR14,p-ERK1/2 in the liver were significantly decreased and no change was observed in the protein levels of ERK1/2 in Simvastatin group and Urantide group. Conclusion :Urantide can inhibit the activation of ERK1/2 by inhibiting the biological effects of UⅡ/GPR14,thus achieving the therapeutic effect on fatty liver.
引文
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[14]Zhao J,Yu Q,Kong W,et al.Theurotens inⅡrecept or antagonist,urantide,protects against atherosclerosis in rats[J].Exp Therapeut Med,2013,5(6):1765-1769.
[15]赵娟,宋成军,蒋菊花,等.Urantide对实验性动脉粥样硬化大鼠肝脏的影响[J].中国老年学杂志,2014,2(34):940-942.
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[18]李学凤,陈军,赵文,等.H-ras12V通过激活SREBP-1c促进肝肿瘤细胞脂肪变性[J].中华肿瘤防治杂志,2015,22(8):579-583.
[19]彭孟云,朱晓宁,汪静.MAPK信号通路与非酒精性脂肪肝关系的研究进展[J].广东医学,2015,36,(5):804-806.
[2]Brancaccio D,Limatola A,Campiglia P,et al.Urantide conformation and interaction with the urotensin-Ⅱreceptor[J].Archiv Der Pharmazie,2014,347(3):185-192.
[3]Ross B,McKendy K,Giaid A,et al.Role of urotensinⅡin health and disease[J].Am J Physiol Regul Integr Comp Physiol,2010,298(5):1156-1172.
[4]Papadopoulos P,Bousette N,Al-Ramli W,et al.Targeted overexpression of the human urotensin receptor transgene in smooth muscle cells[J].Atherosclerosis,2009,204(2):395-404.
[5]Huang Z Y,Yang P Y,Almofti M R,et al.Comparative analysis of the proteome of left ventricular heart of arteriosclerosis in rat[J].Life Sci,2004,75(26):3103-3115.
[6]耿俊梅,万昌武,乐翠云,等.人冠状动脉粥样硬化病灶内CD40L与病灶结构变化的关系[J].中国动脉硬化杂志,2017,25(4):355-359.
[7]Ozturk K,Dogan T,Celikkanat S,et al.The association of fatty pancreas with subclinical atherosclerosis in nonalcoholic fatty liver disease[J].Eur J Gastroenterol Hepatol,2018,30(4):411-417.
[8]Seto W K,Yuen M F.Nonalcoholic fatty liver disease in Asia:emerging perspectives[J].J Gastroenterol,2017,52(2):164-174.
[9]Al-Rasheed N M,Al-Rasheed N M,Hasan I H,et al.Simvastatin ameliorates diabetic cardiomyopathy by attenuating oxidative stress and inflammation in rats[J].Oxid Med Cell Longev,2017,2017:1092015.
[10]莫诗瑶.不同剂量辛伐他汀对减轻急性心肌梗死患者氧化应激反应作用研究[J].白求恩医学杂志,2016,14(6):736-737.
[11]杨红霞,蒋恒波.他汀类药物对老年颈动脉粥样硬化斑块的影响和对急性心脑血管事件的干预作用[J].临床合理用药,2018,11(1A):78-80.
[12]王荣俊,丁波.人urotensinⅡ对大鼠心肌缺血缺氧性损伤的保护作用机制[J].中国临床药理学与治疗学,2017,22(6):662-626.
[13]徐为民,陈欢,孙一平,等.尾加压素Ⅱ受体拮抗剂研究进展[J].中国老年学杂志,2018,38(8):2032-2035.
[14]Zhao J,Yu Q,Kong W,et al.Theurotens inⅡrecept or antagonist,urantide,protects against atherosclerosis in rats[J].Exp Therapeut Med,2013,5(6):1765-1769.
[15]赵娟,宋成军,蒋菊花,等.Urantide对实验性动脉粥样硬化大鼠肝脏的影响[J].中国老年学杂志,2014,2(34):940-942.
[16]Cseh B,Doma E,Baccarini M."RAF"neighborhood:proteinprotein interaction in the Raf/Mek/Erk pathway[J].Febs Lett,2014,588(15):2398-2406.
[17]Watanabe T,Kanome T,Miyazaki A,et al.Human urotensin II as a link between hypertension and coronary artery disease[J].Hypertens Res,2006,29(6):375-387.
[18]李学凤,陈军,赵文,等.H-ras12V通过激活SREBP-1c促进肝肿瘤细胞脂肪变性[J].中华肿瘤防治杂志,2015,22(8):579-583.
[19]彭孟云,朱晓宁,汪静.MAPK信号通路与非酒精性脂肪肝关系的研究进展[J].广东医学,2015,36,(5):804-806.