火绒草对2型糖尿病大鼠腺垂体内NPY和leptin表达的影响
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摘要
目的:观察火绒草对2型糖尿病(T2DM)大鼠腺垂体内神经肽Y(NPY)及leptin表达的影响。方法:将24只SD大鼠随机分为3组,即正常组、模型组和火绒草预处理组。模型组、火绒草预处理组大鼠的建立用链脲佐菌素(STZ)诱导的T2DM大鼠模型,火绒草预处理组于注射STZ前给予火绒草灌胃35 d。大鼠血糖采用酶法检测,大鼠腺垂体内NPY和leptin蛋白及mRNA的表达采用免疫组织化学染色和RT-PCR。结果:火绒草预处理组大鼠血糖明显低于模型组(P<0.01)。免疫组织化学染色显示,NPY和leptin免疫阳性产物均为棕黄色颗粒状,免疫阳性细胞均为腺垂体内嗜色细胞,NPY主要位于细胞质,leptin主要位于细胞核。与模型组大鼠相比,火绒草预处理组大鼠NPY和leptin蛋白的表达明显降低(P<0.01)。RT-PCR结果表明,火绒草预处理组大鼠NPY和leptin mRNA的表达明显低于模型组(P<0.01)。结论:火绒草预处理可下调STZ诱导的大鼠腺垂体内NPY和leptin的高表达,对T2DM腺垂体损伤具有明显保护作用。
Objective: To investigate the effects of Leontopodium alpinum pretreatment on the expressions of neuropeptide Y,NPY) and leptin in the anterior pituitary for rats with STZ-induced type 2 diabetes mellitus( T2DM). Methods:Twenty-four SD rats were randomly divided into 3 groups: control group,model group and Leontopodium alpinum pretreatment group. The modal rats were established by intraperitoneally injection of streptozotocin( STZ) in model group and Leontopodium alpinum pretreatment group. The rats in Leontopodium alpinum pretreatment group were lavaged with Leontopodium alpinum for 35 days before injecting STZ. The enzymic method was used to measure the blood glucose. Immunohistochemical staining and RT-PCR were used to observe the expression of NPY and leptin protein and mRNA,Yespectively in anterior pituitary. Results: The blood glucose of sericine pretreatment group rats were significantly lower than that of model group( P < 0. 01). Immunohistochemical staining showed that NPY and leptin immunoreactive products were brown granular,and expressed in chromophil cells of the anterior pituitary. NPY was located in the cytoplasm,while leptin was mainly located in nucleus. Compared with model rats,the protein expressions of NPY and leptin in the anterior pituitary of the rats in Leontopodium alpinum pretreatment group were decreased obviously( P < 0. 01). RT-PCR results showed that the NPY and leptin mRNA expression in Leontopodium alpinum pretreatment group rats were significantly lower than that in the model group( P < 0. 01). Conclusion: The results show that Leontopodium alpinum pretreatment down-regu-late the high expression of NPY and leptin protein and mRNA in anterior pituitary STZ-induced rats,therefore Leontopodium alpinum has obviously protective effects on anterior pituitary injury for rats with the T2 DM.
Objective: To investigate the effects of Leontopodium alpinum pretreatment on the expressions of neuropeptide Y,NPY) and leptin in the anterior pituitary for rats with STZ-induced type 2 diabetes mellitus( T2DM). Methods:Twenty-four SD rats were randomly divided into 3 groups: control group,model group and Leontopodium alpinum pretreatment group. The modal rats were established by intraperitoneally injection of streptozotocin( STZ) in model group and Leontopodium alpinum pretreatment group. The rats in Leontopodium alpinum pretreatment group were lavaged with Leontopodium alpinum for 35 days before injecting STZ. The enzymic method was used to measure the blood glucose. Immunohistochemical staining and RT-PCR were used to observe the expression of NPY and leptin protein and mRNA,Yespectively in anterior pituitary. Results: The blood glucose of sericine pretreatment group rats were significantly lower than that of model group( P < 0. 01). Immunohistochemical staining showed that NPY and leptin immunoreactive products were brown granular,and expressed in chromophil cells of the anterior pituitary. NPY was located in the cytoplasm,while leptin was mainly located in nucleus. Compared with model rats,the protein expressions of NPY and leptin in the anterior pituitary of the rats in Leontopodium alpinum pretreatment group were decreased obviously( P < 0. 01). RT-PCR results showed that the NPY and leptin mRNA expression in Leontopodium alpinum pretreatment group rats were significantly lower than that in the model group( P < 0. 01). Conclusion: The results show that Leontopodium alpinum pretreatment down-regu-late the high expression of NPY and leptin protein and mRNA in anterior pituitary STZ-induced rats,therefore Leontopodium alpinum has obviously protective effects on anterior pituitary injury for rats with the T2 DM.
引文
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[11]Duwensee K,Schwaiger S,Tancevski I,et al.Leoligin,the major lignan from Edelweiss,activates cholesteryl ester transfer protein[J].Atherosclerosis,2011,219:109-115.
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[14]徐珊,赵明琦,李增强,等.火绒草对大鼠系膜增生性肾小球肾炎的影响[J].中药药理与临床,2013,29:108-111.
[15]柳蕾,匡少金,刘财德,等.火绒草对2型糖尿病大鼠糖脂代谢的影响[J].中国现代医药杂志,2014,16:11-13.
[16]Hornick A,Schwaiger S,Rollinger JM,et al.Extracts and constituents of Leontopodium alpinum enhance cholinergic transmission:brain ACh increasing and memory improving properties[J].Biochem Pharmacol,2008,76:236-248.
[17]Dobner MJ,Sosa S,Schwaiger S,et al.Anti-inflammatory activity of Leontopodium alpinum and its constituents[J].Planta Med,2004,70:502-508.
[18]Schwaiger S,Adams M,Seger C,et al.New constituents of Leontopodium alpinum and their in vitro leukotriene biosynthesis inhibitory activity[J].Planta Med,2004,70:978-985.
[19]Dobner MJ,Schwaiger S,Jenewein IH,et al.Antibacterial activity of Leontopodium alpinum(Edelweiss)[J].J Ethnopharmacol,2003,89:301-303.
[20]李宁川,颜文锦,王金玉.运动训练对糖尿病大鼠糖代谢和下丘脑神经肽Y mRNA的影响[J].中国运动医学杂志,2008,27:52-54.
[21]任艳,李秀钧,田浩明,等.2型糖尿病家系正常糖耐量一级亲属血清瘦素、肿瘤坏死因子-A、神经肽Y水平及与胰岛素抵抗、B细胞分泌功能相关性的临床研究[J].生物医学工程杂志,2010,27:1341-1345.
[22]Dalvi PS,Erbiceanu FD,Irwin DM,et al.Direct regulation of the proglucagon gene by insulin,leptin,and c AMP in embryonic versus adult hypothalamic neurons[J].Mol Endocrino,2012,26:1339-1355.
[23]Nazarians-Armavil A,Menchella JA,Belsham DD.Cellular insulin resistance disrupts leptin-mediated control of neuronal signaling and transcription[J].Mol Endocrinol,2013,27:990-1003.
[24]Lee SJ,Vermas S,Simonds,et al.Leptin stimulates neuropeptide Y and cocaine amphetamine-regulated transcript coexpressing neuronal activity in the dorsomedial hypothalamus in diet-induced obese mice[J].Neuroscience,2013,33:15306-15317.
[2]Gaikwad A,Biju KC,Muthal PL,et al.Role of neuropeptide Y in the regulation of gonadotrop in releasing hormone system in the forebrain of Clarias batrachus(Linn):Immunocytochemistry and high performance liquid chrom atographyelectrosprayionizat ion-mass spectrometric analysis[J].Neuroscience,2005,133:267-279.
[3]Bruinstroop E,Pei L,Ackermans MT,et al.Hypothalamic neuropeptide Y(NPY)controls hepatic VLDL-triglyceride secretion in rats via the sympathetic nervous system[J].Diabetes,2012,61:1043-1050.
[4]马进,张志坚,马瑞,等.2型糖尿病患者血浆神经肽Y、B型钠尿肽、脂联素水平的变化[J].中国老年学杂志,2013,33:2894-2895.
[5]Sksstrand H,Dahlin LB,Lemmark A,et al.Neuropeptide Y and autoantibodies in patients with long-term type 1and type 2 diabetes and neuropathy[J].J Diabetes Complications,2013,27:609-617.
[6]AL-Aazzam SL,Alkhateeb AM,Alzoubi KH,et al.Atorvastatin treatment modulates the interaction between leptin and adiponectin,and the clinical parameters in patients with typeⅡdiabetes[J].Exp Ther Med,2013,6:1565-1569.
[7]Ladyman SR,Fieldwick DM,Grattan DR.Suppression of leptin-induced hypothalamic JAK/STAT signalling and feeding response during pregnancy in the mouse[J].Reproduction,2012,144:83-90.
[8]Wawrosch C,Schwaiger S,Stuppner H,et al.Lignan formation in hairy root cultures of Edelweiss(Leontopodium nivale ssp.alpinum(Cass.)Greuter)[J].Fitoterapia,2014,97:219-23.
[9]Daniela L,Alla P,Maurelli R,et al.Anti-inflammatory effects of concentrated ethanol extracts of Edelweiss(Leontopodium alpinum Cass.)callus cultures towards human keratinocytes and endothelial cells[J].Mediators Inflamm,2012,2012:498373.
[10]Ganzera M,Greifeneder V,Schwaiger S,et al.Chemical profiling of Edelweiss(Leontopodium alpinum Cass.)extracts by micellar electrokinetic capillary chromatography[J].Fitoterapia,2012,83:1680-1686.
[11]Duwensee K,Schwaiger S,Tancevski I,et al.Leoligin,the major lignan from Edelweiss,activates cholesteryl ester transfer protein[J].Atherosclerosis,2011,219:109-115.
[12]Safer S,Cicek SS,Pieri V,et al.Metabolic fingerprinting of Leontopodium species(Asteraceae)by means of 1H NMR and HPLCESI-MS[J].Phytochemistry,2011,72:1379-1389.
[13]Safer S,Tremetsberger K,Guo YP,et al.Phylogenetic relationships in the genus Leontopodium(Asteraceae:Gnaphalieae)based on AFLP data[J].Bot J Linn Soc,2011,165:364-377.
[14]徐珊,赵明琦,李增强,等.火绒草对大鼠系膜增生性肾小球肾炎的影响[J].中药药理与临床,2013,29:108-111.
[15]柳蕾,匡少金,刘财德,等.火绒草对2型糖尿病大鼠糖脂代谢的影响[J].中国现代医药杂志,2014,16:11-13.
[16]Hornick A,Schwaiger S,Rollinger JM,et al.Extracts and constituents of Leontopodium alpinum enhance cholinergic transmission:brain ACh increasing and memory improving properties[J].Biochem Pharmacol,2008,76:236-248.
[17]Dobner MJ,Sosa S,Schwaiger S,et al.Anti-inflammatory activity of Leontopodium alpinum and its constituents[J].Planta Med,2004,70:502-508.
[18]Schwaiger S,Adams M,Seger C,et al.New constituents of Leontopodium alpinum and their in vitro leukotriene biosynthesis inhibitory activity[J].Planta Med,2004,70:978-985.
[19]Dobner MJ,Schwaiger S,Jenewein IH,et al.Antibacterial activity of Leontopodium alpinum(Edelweiss)[J].J Ethnopharmacol,2003,89:301-303.
[20]李宁川,颜文锦,王金玉.运动训练对糖尿病大鼠糖代谢和下丘脑神经肽Y mRNA的影响[J].中国运动医学杂志,2008,27:52-54.
[21]任艳,李秀钧,田浩明,等.2型糖尿病家系正常糖耐量一级亲属血清瘦素、肿瘤坏死因子-A、神经肽Y水平及与胰岛素抵抗、B细胞分泌功能相关性的临床研究[J].生物医学工程杂志,2010,27:1341-1345.
[22]Dalvi PS,Erbiceanu FD,Irwin DM,et al.Direct regulation of the proglucagon gene by insulin,leptin,and c AMP in embryonic versus adult hypothalamic neurons[J].Mol Endocrino,2012,26:1339-1355.
[23]Nazarians-Armavil A,Menchella JA,Belsham DD.Cellular insulin resistance disrupts leptin-mediated control of neuronal signaling and transcription[J].Mol Endocrinol,2013,27:990-1003.
[24]Lee SJ,Vermas S,Simonds,et al.Leptin stimulates neuropeptide Y and cocaine amphetamine-regulated transcript coexpressing neuronal activity in the dorsomedial hypothalamus in diet-induced obese mice[J].Neuroscience,2013,33:15306-15317.