腹腔神经丛阻滞对GK大鼠胰岛素受体底物的影响
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摘要
观察NCPB对GK(Goto-Kakizaki,GK)大鼠血糖及胰岛素受体底物的影响.构建25只非胰岛素依赖糖尿病(NIDDM)的雄性GK大鼠,随机抽取5只GK大鼠,以NCPB前表示,先进行口服葡萄糖耐受试验(OGTT),然后处死采取肝组织.其余20只GK大鼠随机分为对照组和NCPB组,每组10只.NCPB组给予双侧质量分数0.5%利多卡因进行NCPB,1 mL/侧,每天1次,而对照组以质量分数0.9%生理盐水替换.分别在14 d和28 d后取对照组和NCPB组各5只GK大鼠进行OGTT实验,通过Western Blot检测对照组和NCPB组肝组织中的胰岛素受体底物-1(IRS-1)、IRS-2表达以及IRS-1磷酸化情况.结果表明:NCPB组在14 d和28 d NCPB处理后,在0、60和120 min时的血糖均显著低于NCPB前(P<0.05或P<0.01)和对照组(P<0.01),而血清胰岛素水平与NCPB前和对照组相差不显著;在14、28 d NCPB后NCPB组肝组织IRS-1和IRS-2蛋白表达水平与NCPB前相比均显著增加(P<0.01),在相应时间点并显著高于对照组(P<0.01);而对照组的肝组织IRS-1丝氨酸307残基的磷酸化水平在14、28 d NCPB后与NCPB前均有所上调(P<0.05),NCPB组在相应时间点丝氨酸307残基的磷酸化程度均较NCPB前显著下降(P<0.01),且均显著低于对照组(P<0.01).NCPB可降低GK大鼠的空腹血糖,减轻GK大鼠糖耐量受损的程度,其机制可能与NCPB提高胰岛素受体底物的蛋白表达和抑制其丝氨酸磷酸化有关.
To investigate the effects of neurolytic celiac plexus block( NCPB) on blood glucose and insulin receptor substrate in GK( Goto-Kakizaki) rats,the GK rats were used to establish a non-insulin dependent diabetes mellitus( NIDDM) model. 5 GK rats before NCPB were randomly selected to be subjected to the oral glucose tolerance test( OGTT) and the liver tissue was excised. The remaining 20 GK rats were randomly divided into two groups: rats in the NCPB group were treated with 0.5% lidocaine( 1 mL/1 time/day) for 28 consecutive days,while the control group received the same procedure with physiological saline injected instead of 0.5% lidocaine. The OGTT were performed after 14 and 28 days. Western blot assay was used to analyze the expression of insulin receptor substrate 1( IRS-1),IRS-2 and phosphorylation of IRS-1 in the liver tissue. After 14 and 28 days of treatment with NCPB,the concentration of blood glucose in the NCPB group was significantly lower than before NCPB( P<0.05 or P<0.05) and the control group( P<0.01) at 0,60 and 120 min of the OGTT,while the levels of serum insulin exhibited no statistical difference between the control group and the NCPB group. The IRS-1 and IRS-2 expression levels of liver tissue in the NCPB group were significantly higher than control group at the corresponding time points( P<0.01),which significantly increased compared to before NCPB( P<0.01). The phosphorylation level of Ser 307 of IRS-1 was significantly lower in the NCPB group than the control group( P< 0.01) at the corresponding time points,which declined significantly( P<0.01) in the NCPB group,while were significantly increased( P<0.05) in the control group compared with before NCPB. The NCPB treatment can decrease the fasting blood glucose level as well as reduce the degree of impaired glucose tolerance in GK rats. The mechanism may be related to increased protein expression and inhibition of the serine phosphorylation.
To investigate the effects of neurolytic celiac plexus block( NCPB) on blood glucose and insulin receptor substrate in GK( Goto-Kakizaki) rats,the GK rats were used to establish a non-insulin dependent diabetes mellitus( NIDDM) model. 5 GK rats before NCPB were randomly selected to be subjected to the oral glucose tolerance test( OGTT) and the liver tissue was excised. The remaining 20 GK rats were randomly divided into two groups: rats in the NCPB group were treated with 0.5% lidocaine( 1 mL/1 time/day) for 28 consecutive days,while the control group received the same procedure with physiological saline injected instead of 0.5% lidocaine. The OGTT were performed after 14 and 28 days. Western blot assay was used to analyze the expression of insulin receptor substrate 1( IRS-1),IRS-2 and phosphorylation of IRS-1 in the liver tissue. After 14 and 28 days of treatment with NCPB,the concentration of blood glucose in the NCPB group was significantly lower than before NCPB( P<0.05 or P<0.05) and the control group( P<0.01) at 0,60 and 120 min of the OGTT,while the levels of serum insulin exhibited no statistical difference between the control group and the NCPB group. The IRS-1 and IRS-2 expression levels of liver tissue in the NCPB group were significantly higher than control group at the corresponding time points( P<0.01),which significantly increased compared to before NCPB( P<0.01). The phosphorylation level of Ser 307 of IRS-1 was significantly lower in the NCPB group than the control group( P< 0.01) at the corresponding time points,which declined significantly( P<0.01) in the NCPB group,while were significantly increased( P<0.05) in the control group compared with before NCPB. The NCPB treatment can decrease the fasting blood glucose level as well as reduce the degree of impaired glucose tolerance in GK rats. The mechanism may be related to increased protein expression and inhibition of the serine phosphorylation.
引文
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[15]吴峰,都健,王慧敏,等.胰岛素抵抗大鼠的肝脏中丝氨酸P酪氨酸磷酸化异常与血清TNF-α相关[J].中国实验动物学报,2009,17(5):360-363.
[16]HOTAMISLIGI G S,PERALDI P,BUDAVARI A,et al.IRS-1-mediated inhibition of insulin receptor tyrosine kinase activity in TNF-alpha-and obesity-induced insulin resistance[J].Science,1996,271(5249):665-668.
[17]AGUIRRE V,UCHIDA T,YENUSH L,et al.The c-Jun NH(2)-terminal kinase promotes insulin resistance during association with insulin receptor substrate-1 and phosphorylation of Ser(307)[J].J Biol Chem,2000,275(12):9047-9054.
[2]张豫文,洪洁.炎症因子与胰岛素抵抗[J].诊断学理论与实践,2010,9(1):90-94.
[3]SHOELSON S E,LEE J,GOLDFINE A B.Inflammation and insulin resistance[J].J Clin Invest,2006,116(7):1793-1801.
[4]陆建华,粟永萍,程天民,等.颈交感神经阻滞对放烧复合伤小鼠的治疗作用[J].第三军医大学学报,2005,27(12):1253-1255.
[5]YANG F R,WU B S,LAI G H,et al.Assessment of consecutive neurolytic celiac plexus block(NCPB)technique outcomes in the management of refractory visceral cancer pain[J].Pain Med,2012,13(4):518-521.
[6]杨晓春.腹腔神经丛阻滞在慢性腹痛治疗中的应用[J].分子影像学杂志,2015,38(4):393-395.
[7]李娟娥,王磊,秦灵灵,等.自发性2型糖尿病啮齿类动物模型研究概况[J].中国实验方剂学杂志,2010,16(6):267-271.
[8]邹洪,闫洪涛,伍松,等.回肠转位术对自发性糖尿病大鼠肝脏脂代谢的影响及其意义[J].第三军医大学学报,2012,34(11):1114-1116.
[9]陈榕,王莉,曹宏伟,等.糖尿病大鼠肠道、胰腺GLP-1受体mRNA的表达[J].第四军医大学学报,2008,29(13):1235-1238.
[10]姜长林,张丽红,武云飞,等.在体大鼠经背侧入路无水乙醇腹腔神经丛阻滞模型的建立[J].中国疼痛医学杂志,2008,14(4):233-235.
[11]李虹,高奋,边云飞,等.Intermedin对糖尿病大鼠心肌缺血再灌注炎性细胞因子的影响[J].中国动脉硬化杂志,2015,23(10):1005-1011.
[12]CALLE M C,FEMANDEZ M L.Inflammation and type 2 diabetes[J].Diabetes Metab,2012,38(3):183-191.
[13]DONATH M Y.Inflammation as a sensor of metabolic stress in obesity and type 2 diabetes[J].Endocrinology,2011,152(11):4005-4006.
[14]舒适,宋菊敏.胰岛素受体底物-1/-2与胰岛素信号转导[J].医学综述,2008,14(5):723-725.
[15]吴峰,都健,王慧敏,等.胰岛素抵抗大鼠的肝脏中丝氨酸P酪氨酸磷酸化异常与血清TNF-α相关[J].中国实验动物学报,2009,17(5):360-363.
[16]HOTAMISLIGI G S,PERALDI P,BUDAVARI A,et al.IRS-1-mediated inhibition of insulin receptor tyrosine kinase activity in TNF-alpha-and obesity-induced insulin resistance[J].Science,1996,271(5249):665-668.
[17]AGUIRRE V,UCHIDA T,YENUSH L,et al.The c-Jun NH(2)-terminal kinase promotes insulin resistance during association with insulin receptor substrate-1 and phosphorylation of Ser(307)[J].J Biol Chem,2000,275(12):9047-9054.