选取苏钟种母猪16头,在第2个发情期后,按发情前期、发情期、发情后期和间情期随机分成4组。用RT-PCR检测苏钟猪发情周期不同时期preproorexin和orexin 1受体(OX1R) m RNA在下丘脑-垂体-卵巢轴中的表达。结果显示:发情周期不同时期prepro...选取苏钟种母猪16头,在第2个发情期后,按发情前期、发情期、发情后期和间情期随机分成4组。用RT-PCR检测苏钟猪发情周期不同时期preproorexin和orexin 1受体(OX1R) m RNA在下丘脑-垂体-卵巢轴中的表达。结果显示:发情周期不同时期preproorexin m RNA在下丘脑、垂体和卵巢中变化趋势一致,preproorexin m RNA在猪的发情前期表达最高,随后其表达量开始下降,在发情后期其表达最少,在间情期时又开始上升。下丘脑中OX1R m RNA在发情前期开始上升,在发情后期达到最高,随后在间情期又开始下降。垂体和卵巢中OX1 R m RNA的变化与下丘脑中OX1R m RNA的变化趋势一致。上述结果表明:orexin可能参与调控动物生殖过程。展开更多
目的:基于食欲素受体1(OX1R)/磷脂酰肌醇特异性磷酯酶Cβ-1(PLCβ-1)/蛋白激酶Cα(PKCα)/细胞外信号调节激酶1/2(ERK1/2)信号通路探讨安寐丹对失眠大鼠肝脏神经递质及昼夜节律的影响及机制。方法:SPF级SD大鼠60只,随机分为空白组、模...目的:基于食欲素受体1(OX1R)/磷脂酰肌醇特异性磷酯酶Cβ-1(PLCβ-1)/蛋白激酶Cα(PKCα)/细胞外信号调节激酶1/2(ERK1/2)信号通路探讨安寐丹对失眠大鼠肝脏神经递质及昼夜节律的影响及机制。方法:SPF级SD大鼠60只,随机分为空白组、模型组、苏沃雷生组、安寐丹低、中、高剂量组,各10只;除空白组外,其余各组通过腹腔注射对氯苯丙氨酸(PCPA)进行造模,空白组给予等容生理盐水、苏沃雷生组给予苏沃雷生溶液30 mg·kg^(-1)·d^(-1)灌胃、安寐丹低、中、高剂量组分别给予安寐丹水煎液(4.55、9.09、18.18 g·kg^(-1)·d^(-1));观察各组一般情况、体质量和24 h自主活动情况;采用苏木素-伊红(HE)和马松(Masson)染色观察肝脏病理学改变,酶联免疫吸附测定法(ELISA)检测肝脏神经递质γ-氨基丁酸(GABA)、5-羟色胺(5-HT)、肾上腺素(EPI)、去甲肾上腺素(NE)和乙酰胆碱(ACh)的表达,生化检测肝脏谷氨酸(Glu)的表达,实时荧光定量聚合酶链式反应(Real-time PCR)检测肝脏生物钟基因Per1、Per2、Cry1、Cry2、Bmal1、Bmal2的m RNA表达,蛋白免疫印迹法(Western blot)、Real-time PCR检测肝脏OX1R/PLCβ-1/PKCα/ERK1/2信号通路蛋白及m RNA表达。结果:与空白组比较,模型组体质量下降(P<0.05,P<0.01),狂躁、静止节律紊乱(P<0.01),肝脏肌纤维断裂、水肿伴炎性细胞浸润,GABA、5-HT、EPI、NE和ACh含量降低、Glu含量升高(P<0.01),Per1、Per2、Cry1和Cry2 m RNA表达降低(P<0.01),Bmal1和Bmal2 m RNA表达升高(P<0.01),OX1R、PLCβ-1、PKCα、ERK1/2蛋白及m RNA基因表达均增高(P<0.01);与模型组比较,苏沃雷生和安寐丹低、中、高剂量组可增加失眠大鼠体质量(P<0.05,P<0.01),减少狂躁状态、增加其静止时间和频率(P<0.05,P<0.01),并可上调神经递质GABA、5-HT、EPI、NE、ACh和节律基因Per1、Per2、Cry1、Cry2 m RNA表达(P<0.05,P<0.01),抑制Glu及Bmal1、Bmal2、OX1R、PLCβ-1、PKCα、ERK1/2 m RNA和OX1R、PLCβ-1、PKCα、ERK1/2蛋白表达(P<0.05,P<0.01)。结论:安寐丹可通过抑制OX1R/PLCβ-1/PKCα/ERK1/2信号通路调节失眠大鼠肝脏神经递质表达,改善昼夜节律紊乱,且安寐丹高剂量组效果最佳。展开更多
Orexin, released from the hypothalamus, has been implicated in various basic non-somatic functions including feeding, the sleep-wakefulness cycle, emotion,and cognition. However, the role of orexin in somatic motor co...Orexin, released from the hypothalamus, has been implicated in various basic non-somatic functions including feeding, the sleep-wakefulness cycle, emotion,and cognition. However, the role of orexin in somatic motor control is still little known. Here, using whole-cell patch clamp recording and immunostaining, we investigated the effect and the underlying receptor mechanism of orexin-A on neurons in the globus pallidus internus（GPi）,a critical structure in the basal ganglia and an effective target for deep brain stimulation therapy. Our results showed that orexin-A induced direct postsynaptic excitation of GPi neurons in a concentration-dependent manner.The orexin-A-induced excitation was mediated via co-activation of both OX1 and OX2 receptors. Furthermore, the immunostaining results showed that OX1 and OX2 receptors were co-localized in the same GPi neurons. These results suggest that the central orexinergic system actively modulates the motor functions of the basal ganglia via direct innervation on GPi neurons and presumably participates in somatic-non-somatic integration.展开更多
文摘选取苏钟种母猪16头,在第2个发情期后,按发情前期、发情期、发情后期和间情期随机分成4组。用RT-PCR检测苏钟猪发情周期不同时期preproorexin和orexin 1受体(OX1R) m RNA在下丘脑-垂体-卵巢轴中的表达。结果显示:发情周期不同时期preproorexin m RNA在下丘脑、垂体和卵巢中变化趋势一致,preproorexin m RNA在猪的发情前期表达最高,随后其表达量开始下降,在发情后期其表达最少,在间情期时又开始上升。下丘脑中OX1R m RNA在发情前期开始上升,在发情后期达到最高,随后在间情期又开始下降。垂体和卵巢中OX1 R m RNA的变化与下丘脑中OX1R m RNA的变化趋势一致。上述结果表明:orexin可能参与调控动物生殖过程。
文摘目的:基于食欲素受体1(OX1R)/磷脂酰肌醇特异性磷酯酶Cβ-1(PLCβ-1)/蛋白激酶Cα(PKCα)/细胞外信号调节激酶1/2(ERK1/2)信号通路探讨安寐丹对失眠大鼠肝脏神经递质及昼夜节律的影响及机制。方法:SPF级SD大鼠60只,随机分为空白组、模型组、苏沃雷生组、安寐丹低、中、高剂量组,各10只;除空白组外,其余各组通过腹腔注射对氯苯丙氨酸(PCPA)进行造模,空白组给予等容生理盐水、苏沃雷生组给予苏沃雷生溶液30 mg·kg^(-1)·d^(-1)灌胃、安寐丹低、中、高剂量组分别给予安寐丹水煎液(4.55、9.09、18.18 g·kg^(-1)·d^(-1));观察各组一般情况、体质量和24 h自主活动情况;采用苏木素-伊红(HE)和马松(Masson)染色观察肝脏病理学改变,酶联免疫吸附测定法(ELISA)检测肝脏神经递质γ-氨基丁酸(GABA)、5-羟色胺(5-HT)、肾上腺素(EPI)、去甲肾上腺素(NE)和乙酰胆碱(ACh)的表达,生化检测肝脏谷氨酸(Glu)的表达,实时荧光定量聚合酶链式反应(Real-time PCR)检测肝脏生物钟基因Per1、Per2、Cry1、Cry2、Bmal1、Bmal2的m RNA表达,蛋白免疫印迹法(Western blot)、Real-time PCR检测肝脏OX1R/PLCβ-1/PKCα/ERK1/2信号通路蛋白及m RNA表达。结果:与空白组比较,模型组体质量下降(P<0.05,P<0.01),狂躁、静止节律紊乱(P<0.01),肝脏肌纤维断裂、水肿伴炎性细胞浸润,GABA、5-HT、EPI、NE和ACh含量降低、Glu含量升高(P<0.01),Per1、Per2、Cry1和Cry2 m RNA表达降低(P<0.01),Bmal1和Bmal2 m RNA表达升高(P<0.01),OX1R、PLCβ-1、PKCα、ERK1/2蛋白及m RNA基因表达均增高(P<0.01);与模型组比较,苏沃雷生和安寐丹低、中、高剂量组可增加失眠大鼠体质量(P<0.05,P<0.01),减少狂躁状态、增加其静止时间和频率(P<0.05,P<0.01),并可上调神经递质GABA、5-HT、EPI、NE、ACh和节律基因Per1、Per2、Cry1、Cry2 m RNA表达(P<0.05,P<0.01),抑制Glu及Bmal1、Bmal2、OX1R、PLCβ-1、PKCα、ERK1/2 m RNA和OX1R、PLCβ-1、PKCα、ERK1/2蛋白表达(P<0.05,P<0.01)。结论:安寐丹可通过抑制OX1R/PLCβ-1/PKCα/ERK1/2信号通路调节失眠大鼠肝脏神经递质表达,改善昼夜节律紊乱,且安寐丹高剂量组效果最佳。
基金supported by grants from the National Natural Science Foundation of China(NSFC31330033,91332124,31471112,31500848,81671107,31600834,J1210026)+3 种基金the NSFC/Research Grants Council(RGC)Joint Research Scheme(31461163001)the Ministry of Education of China(Specialized Research Fund for the Doctoral Program of Higher Education/RGC Earmarked Research Grant 20130091140003,and Fundamental Research Funds for the Central Universities)the Natural Science Foundation of Jiangsu Province,China(BK2011014,BK20140599,and BK20151384)the Postdoctoral Sciences Foundation,China(2013T60520)
文摘Orexin, released from the hypothalamus, has been implicated in various basic non-somatic functions including feeding, the sleep-wakefulness cycle, emotion,and cognition. However, the role of orexin in somatic motor control is still little known. Here, using whole-cell patch clamp recording and immunostaining, we investigated the effect and the underlying receptor mechanism of orexin-A on neurons in the globus pallidus internus（GPi）,a critical structure in the basal ganglia and an effective target for deep brain stimulation therapy. Our results showed that orexin-A induced direct postsynaptic excitation of GPi neurons in a concentration-dependent manner.The orexin-A-induced excitation was mediated via co-activation of both OX1 and OX2 receptors. Furthermore, the immunostaining results showed that OX1 and OX2 receptors were co-localized in the same GPi neurons. These results suggest that the central orexinergic system actively modulates the motor functions of the basal ganglia via direct innervation on GPi neurons and presumably participates in somatic-non-somatic integration.
