BACKGROUND： The pharmacological action of opioid drugs is related to signal transduction of inhibitory guanine nucleotide binding protein. OBJECTIVE： To quantitatively and qualitatively analyze the influence of morp...BACKGROUND： The pharmacological action of opioid drugs is related to signal transduction of inhibitory guanine nucleotide binding protein. OBJECTIVE： To quantitatively and qualitatively analyze the influence of morphine on levels of type Ⅱ inhibitory guanine nucleotide binding protein （Gi2 protein） in primary cultured hippocampal neurons at different time points. DESIGN, TIME AND SETTING： A randomized controlled study, which was performed at the Department of Neurobiology, Changzheng Hospital, Second Military Medical University of Chinese PLA between September 2002 and March 2004. MATERIALS： Cerebral hippocampal neurons were obtained from newborn SD rats at 1 2 days of age. Biotin-antibody Ⅱ-avidin fluorescein isothiocyanate （Avidin-FITC） was purchased from Sigma Company （USA） and the Gi2 protein polyclonal antibody from Santa Cruz Biochemistry Company （USA）. METHODS： Seven days after culture, mature hippocampal neurons were randomly divided into six groups： 4-, 8-, 16-, 24-, and 48-hour morphine groups, and a blank control group. Neurons in the morphine groups received morphine （10 μ mol/L）, which could cause alterations of G-protein mRNA and cAMP expression in the prefrontal cortex. Neurons in the blank control group were given the same volume of saline. MAIN OUTCOME MEASURES： Gi2 protein levels were detected by an immunofluorescence technique, and were analyzed by the image analytic system with the use of green fluorescence intensity. RESULTS： Gi2 protein levels in hippocampal neurons gradually decreased in the 4-, 8-, 16-, 24-, and 48-hour morphine groups. In particular, Gi2 protein levels in the 16-, 24-, and 48-hour morphine groups were significantly lower than that in the blank control group （P 〈 0.05 0.01）. CONCLUSION： Morphine may decrease Gi2 protein level in primary hippocampal neurons, and the decreasing trend is positively related to morphine-induced time.展开更多
BACKGROUND： Drug addiction involves two main central nervous systems, namely the dopamine and noradrenaline systems. These systems are primarily distributed in five brain regions： the ventral tegmental area, the nuc...BACKGROUND： Drug addiction involves two main central nervous systems, namely the dopamine and noradrenaline systems. These systems are primarily distributed in five brain regions： the ventral tegmental area, the nucleus accumbens, the prefrontal cortex, the hippocampus, and the locus coeruleus. OBJECTIVE： To investigate regional changes of guanine nucleotide binding protein-inhabitant 2 （Gi2） in dopaminergic and noradrenergic neurons in brains of morphine-tolerant and -dependent rats. DESIGN, TIME, AND SETTING： A randomized control study was performed at the Department of Neurobiology in the Second Military Medical University of Chinese PLA （Shanghai, China） between September 2002 and March 2004. MATERIALS： Thirty-six, healthy, male, Sprague-Dawley （SD） rats were used to establish morphine-dependent models. Morphine hydrochloride was a product of Shenyang First Pharmaceutical Factory （China）; naloxone hydrochloride was a product of Beijing Four-Ring Pharmaceutical Factory （China）; and α subunit of Gi2 antibody was offered by Santa Cruz Biotechnology, lnc （USA）. METHODS： Thirty-six SD rats were randomly divided into six groups （n = 6）： （1） acute morphine-dependent group, （2） acute abstinent group, （3） acute control group, （4） chronic morphine-dependent group, （5） chronic abstinent group, and （6） chronic control group. Rats in the acute morphine-dependent and the acute groups were injected with morphine （5 mg/kg）, one injection every two hours, for a total of eight injections. In the acute and chronic morphine-dependent rat models, morphine withdrawal syndrome was precipitated by an injection of naloxone （5 mg/kg）. Rats in the acute control group were given a peritoneal injection of physiological saline at the same administration time as the above two groups. Rats in the chronic morphine-dependent and chronic abstinent groups were injected with morphine three times per day. The administration dose on day 1 was initially 5 mg/kg at 20：00, which increased by 5 mg/kg at 8：00, 12：00, and 20：00 until day 7. On day 13, the dose continuously increased by 10 mg/kg until a chronic morphine-dependent rat model was successfully induced. Afterwards, the rats presented with withdrawal syndromes on naloxone （5 mg/kg） at 8：00 on the same day. Rats in the chronic control group were injected with physiological saline at the same time of the two chronic groups. MAIN OUTCOME MEASURES： The concentration of Gi2 protein in the five brain regions （ventral tegmental area, nucleus accumbens, prefrontal cortex, locus coeruleus, and hippocampus） was detected by immunohistochemistry. RESULTS： In the acute morphine-dependent and acute abstinent groups, Gi2 protein concentration was significantly decreased in the nucleus accumbens, compared to the acute control group （P 〈 0.01）, while no obvious changes were detected in other brain regions. In the chronic morphine-dependent and chronic abstinent groups, Gi2 protein concentration was significantly decreased in the nucleus accumbens, but significantly increased in the locus coeruleus （P 〈 0.01 ） compared to the chronic control group. CONCLUSION： Morphine dependence and tolerance may induce obvious reductions of Gi2 protein levels in the nucleus accumbens of rats. Chronic morphine dependence desensitizes the homologous neurons.展开更多
鸟嘌呤核苷酸结合蛋白q多肽(Guanine nucleotide binding protein q polypeptide,GNAQ)是GTP结合蛋白异源三聚体的组成部分,能够通过结合细胞表面受体介导下游信号通路。最近的研究发现,葡萄酒色斑(Port-wine stains,PWS)中GNAQ p.Arg18...鸟嘌呤核苷酸结合蛋白q多肽(Guanine nucleotide binding protein q polypeptide,GNAQ)是GTP结合蛋白异源三聚体的组成部分,能够通过结合细胞表面受体介导下游信号通路。最近的研究发现,葡萄酒色斑(Port-wine stains,PWS)中GNAQ p.Arg183点突变,可能介导细胞外信号调节激酶(Extracellular signal-regulated kinase,ERK)信号通路。本文对GNAQ基因突变可能介导的下游信号通路,包括丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)、G蛋白信号调节因子5(regulator of G protein signaling 5,RGS5)、蛋白激酶C(protein kinase C,PKC)和YAP等信号通路进行综述。展开更多
目的探讨鸟嘌呤核苷酸交换因子C3G/Rap1酶和鸟嘌呤核苷酸交换因子Dock180/Rac1酶信号通路在卵巢癌浸润中的可能作用。方法 Western blot检测Dock180沉默的卵巢癌细胞SKOV3中C3G的表达,验证上皮性卵巢癌组织中Dock180与C3G的表达相关性;...目的探讨鸟嘌呤核苷酸交换因子C3G/Rap1酶和鸟嘌呤核苷酸交换因子Dock180/Rac1酶信号通路在卵巢癌浸润中的可能作用。方法 Western blot检测Dock180沉默的卵巢癌细胞SKOV3中C3G的表达,验证上皮性卵巢癌组织中Dock180与C3G的表达相关性;免疫组化比较卵巢癌组织中Dock180与C3G的表达趋势;免疫荧光观察SKOV3中Dock180与C3G及它们各自的下游蛋白Rac1/Rap1的定位。结果 Dock180基因沉默的细胞中C3G表达明显增强(P<0.05);Dock180与C3G在卵巢癌组织中的表达呈现相反趋势(P<0.05);C3G/Dock180均主要分布于细胞质,下游效应蛋白Rap1/Rac1在细胞膜和细胞质都有表达,但Rap1以细胞质为主,而Rac1可以伸展至细胞膜及细胞膜皱褶。结论卵巢癌细胞和组织中C3G与Dock180表达呈相反趋势,下游蛋白Rap1与Rac1在细胞内的定位分布差异,可能与C3G/Rap1和Dock180/Rac1信号通路在卵巢肿瘤浸润中的不同作用有关。展开更多
文摘BACKGROUND： The pharmacological action of opioid drugs is related to signal transduction of inhibitory guanine nucleotide binding protein. OBJECTIVE： To quantitatively and qualitatively analyze the influence of morphine on levels of type Ⅱ inhibitory guanine nucleotide binding protein （Gi2 protein） in primary cultured hippocampal neurons at different time points. DESIGN, TIME AND SETTING： A randomized controlled study, which was performed at the Department of Neurobiology, Changzheng Hospital, Second Military Medical University of Chinese PLA between September 2002 and March 2004. MATERIALS： Cerebral hippocampal neurons were obtained from newborn SD rats at 1 2 days of age. Biotin-antibody Ⅱ-avidin fluorescein isothiocyanate （Avidin-FITC） was purchased from Sigma Company （USA） and the Gi2 protein polyclonal antibody from Santa Cruz Biochemistry Company （USA）. METHODS： Seven days after culture, mature hippocampal neurons were randomly divided into six groups： 4-, 8-, 16-, 24-, and 48-hour morphine groups, and a blank control group. Neurons in the morphine groups received morphine （10 μ mol/L）, which could cause alterations of G-protein mRNA and cAMP expression in the prefrontal cortex. Neurons in the blank control group were given the same volume of saline. MAIN OUTCOME MEASURES： Gi2 protein levels were detected by an immunofluorescence technique, and were analyzed by the image analytic system with the use of green fluorescence intensity. RESULTS： Gi2 protein levels in hippocampal neurons gradually decreased in the 4-, 8-, 16-, 24-, and 48-hour morphine groups. In particular, Gi2 protein levels in the 16-, 24-, and 48-hour morphine groups were significantly lower than that in the blank control group （P 〈 0.05 0.01）. CONCLUSION： Morphine may decrease Gi2 protein level in primary hippocampal neurons, and the decreasing trend is positively related to morphine-induced time.
文摘BACKGROUND： Drug addiction involves two main central nervous systems, namely the dopamine and noradrenaline systems. These systems are primarily distributed in five brain regions： the ventral tegmental area, the nucleus accumbens, the prefrontal cortex, the hippocampus, and the locus coeruleus. OBJECTIVE： To investigate regional changes of guanine nucleotide binding protein-inhabitant 2 （Gi2） in dopaminergic and noradrenergic neurons in brains of morphine-tolerant and -dependent rats. DESIGN, TIME, AND SETTING： A randomized control study was performed at the Department of Neurobiology in the Second Military Medical University of Chinese PLA （Shanghai, China） between September 2002 and March 2004. MATERIALS： Thirty-six, healthy, male, Sprague-Dawley （SD） rats were used to establish morphine-dependent models. Morphine hydrochloride was a product of Shenyang First Pharmaceutical Factory （China）; naloxone hydrochloride was a product of Beijing Four-Ring Pharmaceutical Factory （China）; and α subunit of Gi2 antibody was offered by Santa Cruz Biotechnology, lnc （USA）. METHODS： Thirty-six SD rats were randomly divided into six groups （n = 6）： （1） acute morphine-dependent group, （2） acute abstinent group, （3） acute control group, （4） chronic morphine-dependent group, （5） chronic abstinent group, and （6） chronic control group. Rats in the acute morphine-dependent and the acute groups were injected with morphine （5 mg/kg）, one injection every two hours, for a total of eight injections. In the acute and chronic morphine-dependent rat models, morphine withdrawal syndrome was precipitated by an injection of naloxone （5 mg/kg）. Rats in the acute control group were given a peritoneal injection of physiological saline at the same administration time as the above two groups. Rats in the chronic morphine-dependent and chronic abstinent groups were injected with morphine three times per day. The administration dose on day 1 was initially 5 mg/kg at 20：00, which increased by 5 mg/kg at 8：00, 12：00, and 20：00 until day 7. On day 13, the dose continuously increased by 10 mg/kg until a chronic morphine-dependent rat model was successfully induced. Afterwards, the rats presented with withdrawal syndromes on naloxone （5 mg/kg） at 8：00 on the same day. Rats in the chronic control group were injected with physiological saline at the same time of the two chronic groups. MAIN OUTCOME MEASURES： The concentration of Gi2 protein in the five brain regions （ventral tegmental area, nucleus accumbens, prefrontal cortex, locus coeruleus, and hippocampus） was detected by immunohistochemistry. RESULTS： In the acute morphine-dependent and acute abstinent groups, Gi2 protein concentration was significantly decreased in the nucleus accumbens, compared to the acute control group （P 〈 0.01）, while no obvious changes were detected in other brain regions. In the chronic morphine-dependent and chronic abstinent groups, Gi2 protein concentration was significantly decreased in the nucleus accumbens, but significantly increased in the locus coeruleus （P 〈 0.01 ） compared to the chronic control group. CONCLUSION： Morphine dependence and tolerance may induce obvious reductions of Gi2 protein levels in the nucleus accumbens of rats. Chronic morphine dependence desensitizes the homologous neurons.
文摘鸟嘌呤核苷酸结合蛋白q多肽(Guanine nucleotide binding protein q polypeptide,GNAQ)是GTP结合蛋白异源三聚体的组成部分,能够通过结合细胞表面受体介导下游信号通路。最近的研究发现,葡萄酒色斑(Port-wine stains,PWS)中GNAQ p.Arg183点突变,可能介导细胞外信号调节激酶(Extracellular signal-regulated kinase,ERK)信号通路。本文对GNAQ基因突变可能介导的下游信号通路,包括丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)、G蛋白信号调节因子5(regulator of G protein signaling 5,RGS5)、蛋白激酶C(protein kinase C,PKC)和YAP等信号通路进行综述。
文摘目的探讨鸟嘌呤核苷酸交换因子C3G/Rap1酶和鸟嘌呤核苷酸交换因子Dock180/Rac1酶信号通路在卵巢癌浸润中的可能作用。方法 Western blot检测Dock180沉默的卵巢癌细胞SKOV3中C3G的表达,验证上皮性卵巢癌组织中Dock180与C3G的表达相关性;免疫组化比较卵巢癌组织中Dock180与C3G的表达趋势;免疫荧光观察SKOV3中Dock180与C3G及它们各自的下游蛋白Rac1/Rap1的定位。结果 Dock180基因沉默的细胞中C3G表达明显增强(P<0.05);Dock180与C3G在卵巢癌组织中的表达呈现相反趋势(P<0.05);C3G/Dock180均主要分布于细胞质,下游效应蛋白Rap1/Rac1在细胞膜和细胞质都有表达,但Rap1以细胞质为主,而Rac1可以伸展至细胞膜及细胞膜皱褶。结论卵巢癌细胞和组织中C3G与Dock180表达呈相反趋势,下游蛋白Rap1与Rac1在细胞内的定位分布差异,可能与C3G/Rap1和Dock180/Rac1信号通路在卵巢肿瘤浸润中的不同作用有关。
