肥胖大鼠血糖及胰岛素水平与下丘脑中增食欲素受体1的表达
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摘要
目的研究增食欲素受体1(OX1R)反义硫代磷酸化-脱氧寡核苷酸(OX1R-PS-ASDDNs)对肥胖大鼠下丘脑OX1R蛋白表达及血糖、胰岛素代谢的影响。方法 40只大鼠建立高脂饮食诱导的肥胖大鼠模型(肥胖组),37只造模成功,另选择进食普通饲料大鼠10只为正常组。取32只肥胖模型大鼠随机分为反义组、错义组、盐水组、对照组,每组8只,除对照组外,分别于侧脑室插管注射OX1R反义、错义硫代磷酸化-脱氧寡核苷酸及生理盐水。微型血糖仪检测各组大鼠血糖,化学发光免疫法测定血清胰岛素,免疫组化方法分析下丘脑OX1R蛋白的表达。结果肥胖组大鼠体重、Lee's指数、胰岛素、血糖及体脂含量均增加,与正常组比较均有统计学差异(P <0. 01)。下丘脑OX1R蛋白表达肥胖组较正常组增加,差异有统计学意义(P <0. 05)。大鼠侧脑室注射OX1R-PS-ASODNs后,肥胖大鼠反义组较对照组、错义组及盐水组OX1R蛋白表达减少,血糖及胰岛素水平下降,差异有统计学意义(P <0. 05)。结论 OX1R与营养性肥胖相关,参与能量平衡与代谢调节,OX1R-PS-ASODNs可减少营养性肥胖大鼠下丘脑OX1R蛋白的表达,改善血糖及胰岛素水平。
Objective To investigate the effect oforexin receptor 1(OX1R) antisensephosphorothioateoligodeoxynucleotides(OX1R-PS-ASODNs) on the metabolism of blood glucoseand insulinand the expression of OX1R protein in hypothalamus of obese rats. Methods Obese model induced by high-fat diet was established in 40 rats(obesity group),and the obese model was successfully established in 37 rats. Ten rats fed ordinary feed were served as normal control group. Twenty-four rats of 32 obese rats were randomly divided into antisense group,mismatch group,saline group and control group(n = 8 each). Except for control group,OX1R antisense and mismatch phosphorothioateoligodeoxynucleotides(PS-ODNs) and normal saline were respectively given via intracerebroventricular intubation injection in the other 3 groups. Mini glucometer was used to detect blood glucose. Chemiluminescence immunoassay(DPC corporation analyzer) was used to detect serum insulin. Immunohistochemical assay was used to analyze the expression of OX1R protein in hypothalamus. Results Compared with normal control group,body weight,Lee's index and the expression of OX1R protein in hypothalamus in obesity group increased significantly(P < 0. 01,P < 0. 05). After intracerebroventricular injection of OX1R-PS-ASODNs,OX1R protein expression level decreased,and the levels of blood glucose and insulin declined in obesity group compared with control group,mismatch group and saline group(all P <0. 01). Conclusion OX1R is associated with nutritional obesity and participates in energy balance and metabolic regulation,while OX1R-PS-ASODNs can decrease the expression of OX1R protein in hypothalamus and improve blood glucose and insulin levels for the nutritional obesity rats.
Objective To investigate the effect oforexin receptor 1(OX1R) antisensephosphorothioateoligodeoxynucleotides(OX1R-PS-ASODNs) on the metabolism of blood glucoseand insulinand the expression of OX1R protein in hypothalamus of obese rats. Methods Obese model induced by high-fat diet was established in 40 rats(obesity group),and the obese model was successfully established in 37 rats. Ten rats fed ordinary feed were served as normal control group. Twenty-four rats of 32 obese rats were randomly divided into antisense group,mismatch group,saline group and control group(n = 8 each). Except for control group,OX1R antisense and mismatch phosphorothioateoligodeoxynucleotides(PS-ODNs) and normal saline were respectively given via intracerebroventricular intubation injection in the other 3 groups. Mini glucometer was used to detect blood glucose. Chemiluminescence immunoassay(DPC corporation analyzer) was used to detect serum insulin. Immunohistochemical assay was used to analyze the expression of OX1R protein in hypothalamus. Results Compared with normal control group,body weight,Lee's index and the expression of OX1R protein in hypothalamus in obesity group increased significantly(P < 0. 01,P < 0. 05). After intracerebroventricular injection of OX1R-PS-ASODNs,OX1R protein expression level decreased,and the levels of blood glucose and insulin declined in obesity group compared with control group,mismatch group and saline group(all P <0. 01). Conclusion OX1R is associated with nutritional obesity and participates in energy balance and metabolic regulation,while OX1R-PS-ASODNs can decrease the expression of OX1R protein in hypothalamus and improve blood glucose and insulin levels for the nutritional obesity rats.
引文
[1]张靓,赵玉岩,常晓岑,等.增食欲素系统及其功能的研究进展[J].中国糖尿病杂志,2017,25(9):847-850.
[2]Park ES,Yi SJ,Kim JS,et al. Changes in orexin-A and neuropeptide Y expression in the hypothalamus of the fasted and high-fat diet fed rats[J]. J Vet Sci,2004,5(4):295-302.
[3]刘晶,李彩英,高哲,等.青少年儿童糖尿病临床分析[J].河北医药,2013,35(7):987-988.
[4]米成林,贾刚,邓秋红,等.增食欲素A在动物糖脂代谢平衡中的作用和机制[J].动物营养学报,2014,26(9):2491-2498.
[5]Russo F,Pavone LM,Tafuri S,et al. Expression of orexin A and its receptor 1 in the bovine urethroprostatic complex[J]. Anat Rec(Hoboken),2008,291(2):169-174.
[6]郝圆圆,张婷婷,袁红网,等.增食欲素干预单纯性肥胖的作用及机制[J].国际内分泌代谢杂志,2014,34(5):324-326.
[7]Funato H,Tsai AL,Willie JT,et al. Enhanced orexin receptor-2 signaling prevents diet-induced obesity and improves leptin sensitivity[J]. Cell Metab,2009,9(1):64-76.
[8]Inutsuka A,Yamanaka A. The regulation of sleep and wakefulness by the hypothalamic neuropeptide orexin/hypocretin[J]. Nagoya J Med Sci,2013,75(1/2):29-36.
[9]赵琳,贾鲲鹏,王璐,等.胰岛素诱导的低血糖大鼠下丘脑增食欲素A的表达[J].中国应用生理学杂志,2014,30(3):212-213,217.
[10]Russo F,Maruccio L,Calamo A,et al. Orexin 1 receptor in the seminiferous tubules of boar testis:an immunohistochemical study[J].Acta Histochem,2014,116(1):286-288.
[11]常晓岑,赵玉岩,郑德禄,等.增食欲素对肾上腺功能的调节作用及其机制的研究进展[J].中华内分泌代谢杂志,2013,29(5):435-437.
[12]Heinonen MV,Purhonen AK,MkelKA,et al. Functions of orexins in peripheral tissues[J]. Acta Physiol(Oxf),2008,192(4):471-485.
[2]Park ES,Yi SJ,Kim JS,et al. Changes in orexin-A and neuropeptide Y expression in the hypothalamus of the fasted and high-fat diet fed rats[J]. J Vet Sci,2004,5(4):295-302.
[3]刘晶,李彩英,高哲,等.青少年儿童糖尿病临床分析[J].河北医药,2013,35(7):987-988.
[4]米成林,贾刚,邓秋红,等.增食欲素A在动物糖脂代谢平衡中的作用和机制[J].动物营养学报,2014,26(9):2491-2498.
[5]Russo F,Pavone LM,Tafuri S,et al. Expression of orexin A and its receptor 1 in the bovine urethroprostatic complex[J]. Anat Rec(Hoboken),2008,291(2):169-174.
[6]郝圆圆,张婷婷,袁红网,等.增食欲素干预单纯性肥胖的作用及机制[J].国际内分泌代谢杂志,2014,34(5):324-326.
[7]Funato H,Tsai AL,Willie JT,et al. Enhanced orexin receptor-2 signaling prevents diet-induced obesity and improves leptin sensitivity[J]. Cell Metab,2009,9(1):64-76.
[8]Inutsuka A,Yamanaka A. The regulation of sleep and wakefulness by the hypothalamic neuropeptide orexin/hypocretin[J]. Nagoya J Med Sci,2013,75(1/2):29-36.
[9]赵琳,贾鲲鹏,王璐,等.胰岛素诱导的低血糖大鼠下丘脑增食欲素A的表达[J].中国应用生理学杂志,2014,30(3):212-213,217.
[10]Russo F,Maruccio L,Calamo A,et al. Orexin 1 receptor in the seminiferous tubules of boar testis:an immunohistochemical study[J].Acta Histochem,2014,116(1):286-288.
[11]常晓岑,赵玉岩,郑德禄,等.增食欲素对肾上腺功能的调节作用及其机制的研究进展[J].中华内分泌代谢杂志,2013,29(5):435-437.
[12]Heinonen MV,Purhonen AK,MkelKA,et al. Functions of orexins in peripheral tissues[J]. Acta Physiol(Oxf),2008,192(4):471-485.