中药参芪复方对自发性糖尿病GK大鼠大血管Akt与内皮一氧化氮合成酶的影响
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摘要
目的:观察参芪复方对自发性糖尿病(Goto-Kakizaki,GK)大鼠大血管Akt及内皮一氧化氮合成酶(endothelial nitric oxide synthase,e NOS)表达的影响,探讨参芪复方保护糖尿病大血管损伤的机制。方法:采用随机数字表法将73只GK大鼠分为4组:GK空白组(n=18),模型组(n=19),西药阿托伐他汀钙片组(简称西药组,n=18)及中药参芪复方组(简称中药组,n=18)。以18只Wistar大鼠作为对照组,模型组、西药组及中药组大鼠均给予0. 1mg/(ml·d) e NOS抑制剂Nω-L-硝基-精氨酸甲酯(L-NAME)制备糖尿病大血管病变模型。造模同时开始给药,Wistar组、GK及模型组灌服生理盐水5ml/(kg·d);西药组灌服阿托伐他汀钙悬液1. 6mg/(kg·d);中药组灌服中药参芪复方混悬液1. 44g/(kg·d),对照组喂饲普通饲料,其余各组喂饲高脂饲料。干预35天后处死大鼠,光镜观察各组大鼠主动脉形态,实时荧光定量PCR法测定各组大鼠腹主动脉Akt m RNA及e NOS m RNA表达量,采用肯德尔秩相关检验分析主动脉血管壁Akt、e NOS m RNA表达水平的相关性。结果:与模型组比较,中药组与西药组腹主动脉内膜的病变程度及病变数目均有显著性差异(P<0. 05),而中药组与西药组腹主动脉内膜病变数目及程度比较,差异无统计学意义(P>0. 05);与模型组比较,中药组与西药组腹主动脉血管壁Akt m RNA及e NOS m RNA表达水平明显降低(P<0. 01),且两组Akt m RNA与e NOS m RNA表达均呈正相关(P<0. 05)。结论:参芪复方能有效延缓GK大鼠的大血管病变的发生和发展,其作用机制可能与抑制大血管壁Akt及e NOS的表达有关。
Objective: To observe the influence of Akt and e NOS in the diabetes mellitus GK rat abdominal aorta vessel wall with Shenqi Compound Recipe( SQCR). And to investigate SQCR's protective mechanisms on large-vessel's injury. Methods: 73 GK male rats were randomly divided into four groups with random number. Methods: 18 in blank group,19 in model group,18 in western medicine group and Chinese medicine group respectively. Except for blank group,the other three groups were administered the Nù-nitro-L-arginine methyl ester( L-NAME) preparing for the diabetes vasculopathy model. At the same time,the blank group and the model group were given NS,the western medicine group were given atorvastatin and the Chinese medicine group given SQCR. The blank group with normal feed and the other three groups with high fat diet. The rats in 4 groups were killed35 days later. The morphology of aorta in each group was observed by light microscope and m RNA content of Akt and e NOS in abdominal aorta were detected by Real-time PCR,the correlationship analysis of Akt and e NOS expression in aortic wall by Kendall rank correlationship test. Results: Compared with the model group,the number of the abdominal aorta intima lesion and degree difference in the western medicine group and the chinese medicine groupthe( P < 0. 05),but there was no significant difference between the western medicine group and the Chinese medicine group( P > 0. 05); Compared with the model group,Akt and e NOS m RNA expression in the aorta vessel wall in the western medicine group and the Chinese medicine group were significantly decreased( P < 0. 01),and Akt and e NOS m RNA expression in the two groups were positively correlated( P < 0. 05). Conclusion: SQCR can delay the macroangiopathy in the spontaneous DM GK rats. The mechanism may be related to the depression of Akt and e NOS expression in the aorta vessel wall.
Objective: To observe the influence of Akt and e NOS in the diabetes mellitus GK rat abdominal aorta vessel wall with Shenqi Compound Recipe( SQCR). And to investigate SQCR's protective mechanisms on large-vessel's injury. Methods: 73 GK male rats were randomly divided into four groups with random number. Methods: 18 in blank group,19 in model group,18 in western medicine group and Chinese medicine group respectively. Except for blank group,the other three groups were administered the Nù-nitro-L-arginine methyl ester( L-NAME) preparing for the diabetes vasculopathy model. At the same time,the blank group and the model group were given NS,the western medicine group were given atorvastatin and the Chinese medicine group given SQCR. The blank group with normal feed and the other three groups with high fat diet. The rats in 4 groups were killed35 days later. The morphology of aorta in each group was observed by light microscope and m RNA content of Akt and e NOS in abdominal aorta were detected by Real-time PCR,the correlationship analysis of Akt and e NOS expression in aortic wall by Kendall rank correlationship test. Results: Compared with the model group,the number of the abdominal aorta intima lesion and degree difference in the western medicine group and the chinese medicine groupthe( P < 0. 05),but there was no significant difference between the western medicine group and the Chinese medicine group( P > 0. 05); Compared with the model group,Akt and e NOS m RNA expression in the aorta vessel wall in the western medicine group and the Chinese medicine group were significantly decreased( P < 0. 01),and Akt and e NOS m RNA expression in the two groups were positively correlated( P < 0. 05). Conclusion: SQCR can delay the macroangiopathy in the spontaneous DM GK rats. The mechanism may be related to the depression of Akt and e NOS expression in the aorta vessel wall.
引文
[1]中华医学会糖尿病学分会糖尿病慢性并发症调查组.全国住院糖尿病患者慢性并发症及其相关危险因素10年回顾性调查分析[J].中华糖尿病杂志,2003,11(4):5~10
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[8]庄灿,谢春光,陈敏,等.参芪复方对GK大鼠主动脉血管紧张素Ⅱ1型受体m RNA表达的影响[J].中国中西医结合杂志,2013,33(3):351~356
[9]刘桠,谢春光,庄灿,等.参芪复方对GK大鼠2型糖尿病大血管病变内皮保护作用的实验研究[J].辽宁中医杂志,2010,37(6):1163~1165
[10]刘桠,谢春光,陈敏,等.参芪复方调控GK大鼠大血管病变PTEN/PI3K通路的实验研究[J].中国中西医结合杂志,2010,30(6):640~644
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[12]林兰主编.现代中医糖尿病学[M].北京:人民卫生出版社,2008:737~739
[13] Matsuura E.,et al. Oxidized LDL/beta2-glycoprotein I complexes:new aspects in atherosclerosis[J]. Lupus,2005,14(9):736~741
[14] Fulton D,Gratton JP.,Mc Cabe TJ,et al. Regulation of endotuelium-derived nitric oxide production by the protein kinase AKT[J].Nature,1999,399:597~601
[15] Jones PF,Jakubowicz T,Pitossi FJ,et al. Molecular cloning and identification of a serine/threonine protein kinase of the second-messenger subfamily[J]. Proc Natl Acad Sci U S A,1991,88(10):4171~4175
[16] Coffer PJ,Woodgett JR. Molecular cloning and characterization of a novel putative protein-serine kinase related to the c AMP-dependent and protein kinase C families. Eur J Biochem,1991,201(2):475~481
[17] Rajala MS,Rajala RV,Astley RA,et al. Corneal Cell Survival in Adenovirus Type 19Infection Requires Phosphoinositide 3-Kinase/Akt Activation[J]. J Virol,2005,79(19):12332~12341
[18] Tong Q,Zheng L,Lin L,et al. Hypoxia-induced mitogenic factor promotes vascular adhesion molecule-1expression via the PI3K/AktNF-kappaB signaling pathway[J]. Am J Respir Cell Mol Biol,2006,35(4):444~456
[19] Hu Z,Xiong Y,Han X,et al. Acute mechanical stretch promotes e NOS activation in venous endothelial cells mainly via PKA and Akt pathways[J]. PloS one,2013,8(8):e71359
[20] Haeussler DJ,Pimentel DR,Hou X,et al. Endomembrane H-Ras controls vascular endothelial growth factor-induced nitric-oxide synthasE-mediated endothelial cell migration[J]. J Biol Chem,2013,288(21):15380~15389
[21] Everaert BR,Van Craenenbroeck EM,Hoymans VY,et al.Current perspective of pathophysiological and interventional effects on endothelial progenitor cell biology:focus on PI3K/AKT/e NOS pathway[J]. Intern J Cardiol,2010,144(3):350~366
[22] Yki-Jarvinen,et al. Best Pract Res Clin Endocrinol Metab,2003,17:411~430
[23] Fulvio Morello,Alessia Perino,Emilio Hirsch. Phosphoinositide 3-kinase signaling in the vascular system[J]. Cardiovascular Research,2008,18:1~11
[2]中华医学会糖尿病分会.中国2型糖尿病防治指南2010版[M].北京:北京大学医学出版社,2011:5~7
[3]谢毅强,李军茹,张红敏,等.参芪复方治疗2型糖尿病胰岛素抵抗的临床研究[J].中华实用中西医杂志,2005,18(17):844~846
[4]王芬,何华亮,张红敏,等.参芪复方对GK大鼠脂代谢异常的实验研究[J].天津中医药,2007,24(6):507~509
[5]张红敏,陈世伟,谢春光,等.参芪复方抗自发性糖尿病GK大鼠早期动脉粥样硬化的作用机制[J].中国中药杂志,2006,31(15):1272~1277
[6]殷丽平,杜联,谢春光,等.参芪复方对实验性2型糖尿病大血管病变胰岛素抵抗的干预作用[J].成都中医药大学学报,2010,33(3):54~56
[7]田辉,樊柏林,刘瑶,等. SPF级Wistar大鼠血液学及血生化指标正常值范围探讨[J].实验动物学,2007,24(2):1~4
[8]庄灿,谢春光,陈敏,等.参芪复方对GK大鼠主动脉血管紧张素Ⅱ1型受体m RNA表达的影响[J].中国中西医结合杂志,2013,33(3):351~356
[9]刘桠,谢春光,庄灿,等.参芪复方对GK大鼠2型糖尿病大血管病变内皮保护作用的实验研究[J].辽宁中医杂志,2010,37(6):1163~1165
[10]刘桠,谢春光,陈敏,等.参芪复方调控GK大鼠大血管病变PTEN/PI3K通路的实验研究[J].中国中西医结合杂志,2010,30(6):640~644
[11]高泓,谢春光,赵旭.糖尿病大血管病变的中医病机分析[J].时珍国医国药,2009,20(6):1314~1315
[12]林兰主编.现代中医糖尿病学[M].北京:人民卫生出版社,2008:737~739
[13] Matsuura E.,et al. Oxidized LDL/beta2-glycoprotein I complexes:new aspects in atherosclerosis[J]. Lupus,2005,14(9):736~741
[14] Fulton D,Gratton JP.,Mc Cabe TJ,et al. Regulation of endotuelium-derived nitric oxide production by the protein kinase AKT[J].Nature,1999,399:597~601
[15] Jones PF,Jakubowicz T,Pitossi FJ,et al. Molecular cloning and identification of a serine/threonine protein kinase of the second-messenger subfamily[J]. Proc Natl Acad Sci U S A,1991,88(10):4171~4175
[16] Coffer PJ,Woodgett JR. Molecular cloning and characterization of a novel putative protein-serine kinase related to the c AMP-dependent and protein kinase C families. Eur J Biochem,1991,201(2):475~481
[17] Rajala MS,Rajala RV,Astley RA,et al. Corneal Cell Survival in Adenovirus Type 19Infection Requires Phosphoinositide 3-Kinase/Akt Activation[J]. J Virol,2005,79(19):12332~12341
[18] Tong Q,Zheng L,Lin L,et al. Hypoxia-induced mitogenic factor promotes vascular adhesion molecule-1expression via the PI3K/AktNF-kappaB signaling pathway[J]. Am J Respir Cell Mol Biol,2006,35(4):444~456
[19] Hu Z,Xiong Y,Han X,et al. Acute mechanical stretch promotes e NOS activation in venous endothelial cells mainly via PKA and Akt pathways[J]. PloS one,2013,8(8):e71359
[20] Haeussler DJ,Pimentel DR,Hou X,et al. Endomembrane H-Ras controls vascular endothelial growth factor-induced nitric-oxide synthasE-mediated endothelial cell migration[J]. J Biol Chem,2013,288(21):15380~15389
[21] Everaert BR,Van Craenenbroeck EM,Hoymans VY,et al.Current perspective of pathophysiological and interventional effects on endothelial progenitor cell biology:focus on PI3K/AKT/e NOS pathway[J]. Intern J Cardiol,2010,144(3):350~366
[22] Yki-Jarvinen,et al. Best Pract Res Clin Endocrinol Metab,2003,17:411~430
[23] Fulvio Morello,Alessia Perino,Emilio Hirsch. Phosphoinositide 3-kinase signaling in the vascular system[J]. Cardiovascular Research,2008,18:1~11