脾阳虚大鼠下丘脑和小肠缩胆囊素信号转导途径的变化研究
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摘要
目的:通过检测下丘脑和小肠组织缩胆囊素(Cholecystokinin,CCK)信号转导途径的变化,研究脾阳虚的发生机制。方法:16只SD大鼠随机分为对照组和脾阳虚组(模型组),每组8只; ELISA法检测下丘脑和小肠组织CCK-8、环-磷酸腺苷(cycle monophosphate,c AMP)和蛋白激酶A(protein kinase A,PKA)含量;免疫组化法检测下丘脑CCK2受体(CCK 2 receptor,CCK2R)和小肠CCK1R表达。结果:与对照组相比,下丘脑和小肠CCK-8水平明显升高,而下丘脑的CCKR2表达、c AMP和PKA水平以及小肠CCKR1表达和c AMP水平均显著下降,但小肠PKA水平没有明显变化。结论:脾阳虚的发生与下丘脑和小肠组织中CCK过度释放有关。
Objective: To explore the mechanism of spleen Yang deficiency focusing on cholecycstokinin(CCK) transmembrane signal pathway. Methods: Sixteen SD rats were randomly divided into the control and spleen Yang deficiency groups,8 rats in each group. ELISA was used to check CCK-8,c AMP and PKA concentrations in hypothalamic and small intestine(SI). The immunohistochemistry was used to detect CCK 2 receptor(CCK2 R) in hypothalamic tissue and CCK1 R in SI tissues. Results: Compared with those in the control group,CCK-8 levels in hypothalamic and SI tissues were increased. The hypothalamic CCK2 R expression and levels of c AMP and PKA,and SI CCK1 R expression and c AMP level were all reduced,but there was no overt change in SI PKA level in the model group. Conclusion: Overrelease of CCK in hypothalamus and SI tissues might be contribute to the development of spleen Yang deficiency.
Objective: To explore the mechanism of spleen Yang deficiency focusing on cholecycstokinin(CCK) transmembrane signal pathway. Methods: Sixteen SD rats were randomly divided into the control and spleen Yang deficiency groups,8 rats in each group. ELISA was used to check CCK-8,c AMP and PKA concentrations in hypothalamic and small intestine(SI). The immunohistochemistry was used to detect CCK 2 receptor(CCK2 R) in hypothalamic tissue and CCK1 R in SI tissues. Results: Compared with those in the control group,CCK-8 levels in hypothalamic and SI tissues were increased. The hypothalamic CCK2 R expression and levels of c AMP and PKA,and SI CCK1 R expression and c AMP level were all reduced,but there was no overt change in SI PKA level in the model group. Conclusion: Overrelease of CCK in hypothalamus and SI tissues might be contribute to the development of spleen Yang deficiency.
引文
[1]丛培玮,尚冰,王艳杰,等.脾气虚和脾阳虚模型大鼠脑肠肽与下丘脑葡萄糖转运体1及葡萄糖转运体3表达水平变化的实验研究[J].中国全科医学,2016,19(18):2201-2205.
[2]马祥雪,王凤云,符竣杰,等.从脑肠互动角度探讨脾主运化的物质基础与科学内涵[J].中医杂志,2016,57(12):996-999.
[3]余跃,刘旭东,刘文俊,等.血管活性肠肽在脾气虚大鼠小肠运动中的作用[J].时珍国医国药,2015,26(9):2284-2285.
[4]田茸,巩子汉,杨晓轶,等.脾气虚证大鼠脑肠肽动态变化及四君子汤干预效应研究[J].中国中医药信息杂志,2015,22(9):68-71.
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[6]Nomoto S,Ohta M,Kanai S,et al.Absence of the cholecystokinin-Areceptor deteriorates homeostasis of body temperature in response to changes in ambient temperature[J].Am J Physiol Regul Integr Comp Physiol,2004,287(3):R556-561.
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[11]吴德坤,黄瑞诚,郑景辉,等.脾虚大鼠血清及胃组织中胃肠激素GAS、CCK、SS变化及中药干预效果[J].世界华人消化杂志,2015,23(11):1791-1795.
[12]邵南齐,高青,朱萱萱,等.运脾温阳颗粒对脾虚泄泻大鼠胃黏膜结构及血浆中CCK、PP浓度的影响[J].江苏中医药,2014,46(9):75-77.
[13]孙晓芬,程卫东,段永强,等.脾虚大鼠血清及胃组织中胃肠激素SS、GAS和CCK的变化及中药干预研究[J].时珍国医国药,2014,25(3):756-758.
[14]Akieda-Asai S,Poleni PE,Date Y.Coinjection of CCK and leptin reduces food intake via increased CART/TRH and reduced AMPKphosphorylation in the hypothalamus[J].Am J Physiol Endocrinol Metab,2014,306(11):E1284-1291.
[15]Lo CC,Davidson WS,Hibbard SK,et al.Intraperitoneal CCK and fourth-intraventricular Apo AIV require both peripheral and NTSCCK1R to reduce food intake in male rats[J].Endocrinology,2014,155(5):1700-1707.
[16]Kagebayashi T,Kontani N,Yamada Y,et al.Novel CCK-dependent vasorelaxing dipeptide,Arg-Phe,decreases blood pressure and food intake in rodents[J].Mol Nutr Food Res.,2012,56(9):1456-1463.
[17]Szelényi Z.Neuronal CCK and thermoregulation:two receptors with different functions[J].Am J Physiol Regul Integr Comp Physiol.,2007,292(1):R109-111.
[18]Szelényi Z.Cholecystokinin:role in thermoregulation and other aspects of energetics[J].Clin Chim Acta,2010,411(5-6):329-335.
[19]Dodd GT,Worth AA,Nunn N,et al.The thermogenic effect of leptin is dependent on a distinct population of prolactin-releasing peptide neurons in the dorsomedial hypothalamus[J].Cell Metab,2014,20(4):639-649.
[2]马祥雪,王凤云,符竣杰,等.从脑肠互动角度探讨脾主运化的物质基础与科学内涵[J].中医杂志,2016,57(12):996-999.
[3]余跃,刘旭东,刘文俊,等.血管活性肠肽在脾气虚大鼠小肠运动中的作用[J].时珍国医国药,2015,26(9):2284-2285.
[4]田茸,巩子汉,杨晓轶,等.脾气虚证大鼠脑肠肽动态变化及四君子汤干预效应研究[J].中国中医药信息杂志,2015,22(9):68-71.
[5]BalaskóM,Rostás I,Füredi N,et al.Age and nutritional state influence the effects of cholecystokinin on energy balance[J].Exp Gerontol.,2013,48(11):1180-1188.
[6]Nomoto S,Ohta M,Kanai S,et al.Absence of the cholecystokinin-Areceptor deteriorates homeostasis of body temperature in response to changes in ambient temperature[J].Am J Physiol Regul Integr Comp Physiol,2004,287(3):R556-561.
[7]Harper EA,Roberts SP,Kalindjian SB.Thermodynamic analysis of ligands at cholecystokinin CCK2 receptors in rat cerebral cortex[J].Br JPharmacol,2007,151(8):1352-1367.
[8]El-Salhy M,Hatlebakk JG,Hausken T.Reduction in duodenal endocrine cells in irritable bowel syndrome is associated with stem cell abnormalities[J].World J Gastroenterol,2015,21(32):9577-9587.
[9]刘洋,刘旭东,刘文俊,等.基于脾气虚的脾阳虚大鼠模型的复制方法及评价标准研究[J].中国医学创新,2016,13(3):25-28.
[10]赵璠,刘旭东,刘文俊,等.M2和M3型胆碱能受体在脾气虚大鼠小肠运动中的作用[J].中国中医基础医学杂志,2016,22(1):41-43.
[11]吴德坤,黄瑞诚,郑景辉,等.脾虚大鼠血清及胃组织中胃肠激素GAS、CCK、SS变化及中药干预效果[J].世界华人消化杂志,2015,23(11):1791-1795.
[12]邵南齐,高青,朱萱萱,等.运脾温阳颗粒对脾虚泄泻大鼠胃黏膜结构及血浆中CCK、PP浓度的影响[J].江苏中医药,2014,46(9):75-77.
[13]孙晓芬,程卫东,段永强,等.脾虚大鼠血清及胃组织中胃肠激素SS、GAS和CCK的变化及中药干预研究[J].时珍国医国药,2014,25(3):756-758.
[14]Akieda-Asai S,Poleni PE,Date Y.Coinjection of CCK and leptin reduces food intake via increased CART/TRH and reduced AMPKphosphorylation in the hypothalamus[J].Am J Physiol Endocrinol Metab,2014,306(11):E1284-1291.
[15]Lo CC,Davidson WS,Hibbard SK,et al.Intraperitoneal CCK and fourth-intraventricular Apo AIV require both peripheral and NTSCCK1R to reduce food intake in male rats[J].Endocrinology,2014,155(5):1700-1707.
[16]Kagebayashi T,Kontani N,Yamada Y,et al.Novel CCK-dependent vasorelaxing dipeptide,Arg-Phe,decreases blood pressure and food intake in rodents[J].Mol Nutr Food Res.,2012,56(9):1456-1463.
[17]Szelényi Z.Neuronal CCK and thermoregulation:two receptors with different functions[J].Am J Physiol Regul Integr Comp Physiol.,2007,292(1):R109-111.
[18]Szelényi Z.Cholecystokinin:role in thermoregulation and other aspects of energetics[J].Clin Chim Acta,2010,411(5-6):329-335.
[19]Dodd GT,Worth AA,Nunn N,et al.The thermogenic effect of leptin is dependent on a distinct population of prolactin-releasing peptide neurons in the dorsomedial hypothalamus[J].Cell Metab,2014,20(4):639-649.