目的：探讨脂质运载蛋白型前列腺素D合成酶（Lipocalin-type prostaglandin D syn-thase，L-PGDS）在匹罗卡品诱导的癫痫大鼠海马脑组织中的表达变化及意义。方法：35只SD雄性大鼠随机分成对照组和模型组，模型组依据观察时间点不同又...目的：探讨脂质运载蛋白型前列腺素D合成酶（Lipocalin-type prostaglandin D syn-thase，L-PGDS）在匹罗卡品诱导的癫痫大鼠海马脑组织中的表达变化及意义。方法：35只SD雄性大鼠随机分成对照组和模型组，模型组依据观察时间点不同又分成建模后1d、3d、7d、14d、30d和60d6个亚组，每组各5只大鼠。模型组大鼠用氯化锂-匹罗卡品诱导癫痫持续状态后，用蛋白免疫印迹检测大鼠海马组织内L-PGDS蛋白表达水平，并用免疫荧光双标技术对L-PGDS进行海马区细胞定位。结果：大鼠癫痫持续状态后第1天，海马组织内L-PGDS蛋白表达水平较对照组有所升高，随着时间的延长，L-PGDS蛋白表达水平逐渐升高，与对照组比较差异有统计学意义（P〈0．05）。免疫荧光双标检测发现L-PGDS主要表达于神经元的树突和星型胶质细胞的胞质胞浆中。结论：L-PGDS在癫痫模型中表达异常升高，可能参与了癫痫的发生发展过程，有望成为临床癫痫诊断的一个分子标志物和新的抗癫痫药物治疗的靶点。展开更多
本研究旨在克隆犏牛前列腺素D合成酶(prostagland D synthase,L-PGDS)基因序列,并检测其在不同组织及不同发育时期睾丸中的表达水平,从而为深入研究L-PGDS基因在犏牛睾丸发育过程中的作用机制提供依据。采集成年健康犏牛心脏、肝脏、脾...本研究旨在克隆犏牛前列腺素D合成酶(prostagland D synthase,L-PGDS)基因序列,并检测其在不同组织及不同发育时期睾丸中的表达水平,从而为深入研究L-PGDS基因在犏牛睾丸发育过程中的作用机制提供依据。采集成年健康犏牛心脏、肝脏、脾脏、肺脏、肾脏、大肠、小肠、胃、大脑及不同发育时期犏牛睾丸组织:胎牛期(5~6月)、幼年期(1~2岁)、青春期(2.5~4岁)、老年期(7~9岁),Trizol法提取各组织总RNA,利用RT-PCR技术扩增、克隆L-PGDS基因序列并利用相关生物学软件进行分析,利用实时荧光定量PCR检测犏牛各组织及4个不同发育阶段睾丸中L-PGDS基因mRNA的表达水平。结果显示,L-PGDS基因序列全长624 bp,包括完整的开放阅读框576 bp,编码191个氨基酸。同源性比对发现,犏牛与黄牛和绵羊的同源性最高,分别为99.28%和94.0%。系统进化树分析结果显示,犏牛与黄牛亲缘关系最近,其次是绵羊、马。L-PGDS蛋白为不稳定疏水蛋白,分子质量为21.229 ku,分子式为C953H1471N251O281S9,等电点为6.43,不稳定指数为47.25,不含跨膜区及信号肽,蛋白质二级结构预测显示α-螺旋、无规则卷曲、延伸链分别占25.65%、53.40%和20.94%。实时荧光定量PCR检测结果显示,L-PGDS基因在犏牛睾丸组织中特异性高表达,极显著高于其他组织(P<0.01);随着年龄增长L-PGDS基因mRNA在犏牛睾丸中呈先上升后下降趋势,其中在青春期相对表达量最高,极显著高于其他时期(P<0.01)。本研究结果为深入探究L-PGDS基因在犏牛睾丸发育过程中的作用机制提供了基础资料。展开更多
Background： The purpose of this study was to determine the association of three proteins involved in sperm function on the freezability of porcine semen： the heat shock protein 90 alpha（HSP90a）, the Niemann-Pick d...Background： The purpose of this study was to determine the association of three proteins involved in sperm function on the freezability of porcine semen： the heat shock protein 90 alpha（HSP90a）, the Niemann-Pick disease type C2 protein（NPC2）, and lipocalin-type prostaglandin D synthase（L-PGDS）.Six adult boars（each boar was ejaculated three times, 18 in total） were classified by freezability based on the percentage of functionally competent sperm. The male semen with highest freezability（MHF） and the male semen with lowest freezability（MLF） were centrifuged immediately after collection to separate seminal plasma and spermatozoa to make four possible combinations of these two components and to incubate them for 3 h,adjusting the temperature to 17 °C, to freeze them afterwards. The quantification of proteins was performed in two stages： at zero and at 3 h after incubation of the four combinations.Results： The spermatozoa × incubation time（IT） interaction only had effect（P 〈 0.01） on HSP90 a levels; this protein increased in seminal plasma, after 3 h of incubation, in larger quantity（P 〈 0.05） in combinations with MLF spermatozoa. In relation with the NPC2 protein, two isoforms of 16 and 19 kDa were identified. The 19 kDa isoform was affected（P 〈 0.01） only by the seminal plasma × IT interaction, with superior values（P 〈 0.01） both at zero and three hours of incubation, in the combinations with MHF seminal plasma; and 16 kDa isoform was affected（P 〈 0.01） only by the IT with reduction after 3 h of incubation. The levels of L-PGDS was affected（P 〈 0.01）only by the spermatozoa × IT interaction, which reduced（P 〈 0.01） in combinations with MLF spermatozoa after 3 h of incubation.Conclusions： It is possible to consider that the three proteins evaluated were associated with freezability of boar semen due, especially, to the fact that mixtures with MLF spermatozoa showed greater increase levels of the HSP90 a protein and reduction of L-PGDS in plasma. In addition, the seminal plasma of MHF had higher concentration of the NPC2 of 19 kDa protein, which was reduced by incubating with MHF spermatozoa.展开更多
Objectives: Alterations in the prostaglandin-D-system have been found in animal sleep experiments and disorders that present with hypersomnia or sleep disturbances. The recently demonstrated involvement of the leptome...Objectives: Alterations in the prostaglandin-D-system have been found in animal sleep experiments and disorders that present with hypersomnia or sleep disturbances. The recently demonstrated involvement of the leptomeningeal lipocalin- type prostaglandin-Dsynthase (L-PGDS) (β-trace) in human physiological sleep encouraged us to investigate its role in the pathophysiology of narcolepsy. Methods: In a pilot study, serum L-PGDS and melatonin concentrations were assessed in 14 narcoleptic patients during undisturbed sleep and total sleep deprivation, compared with those from 14 healthy controls during undisturbed sleep. Excessive daytime sleepiness was measured by a standardized questionnaire (Epworth slee piness scale, ESS). Results: In narcoleptic patients, markedly increased baseline L-PGDS levels were significantly correlated with the ESS score, but not with the degree of cataplexy. Serum L-PGDS concentrations in patients as well as in controls followed a time-dependent fluctuation with evening increases, highest values during the night and in the morning. Compared with controls, patients exhibited significant/increased amplitude of circulating L-PGDS without any suppression by total sleep deprivation. Conclusion: These findings indicate that the prostaglandin-D-system contributes to the pathophysiology of narcolepsy, e. g. the regulation of excessive daytime sleepiness. Since it has been suggested that L-PGDS is also involved in neurodegenerative disorders, there may be a more specific role of the prostaglandin-D-sys- tem in narcoleptic aetiogenesis. Moreover, its linkage with the immune system as well as with human sleep regulation offers a direct access for investigating both systems.展开更多
文摘目的：探讨脂质运载蛋白型前列腺素D合成酶（Lipocalin-type prostaglandin D syn-thase，L-PGDS）在匹罗卡品诱导的癫痫大鼠海马脑组织中的表达变化及意义。方法：35只SD雄性大鼠随机分成对照组和模型组，模型组依据观察时间点不同又分成建模后1d、3d、7d、14d、30d和60d6个亚组，每组各5只大鼠。模型组大鼠用氯化锂-匹罗卡品诱导癫痫持续状态后，用蛋白免疫印迹检测大鼠海马组织内L-PGDS蛋白表达水平，并用免疫荧光双标技术对L-PGDS进行海马区细胞定位。结果：大鼠癫痫持续状态后第1天，海马组织内L-PGDS蛋白表达水平较对照组有所升高，随着时间的延长，L-PGDS蛋白表达水平逐渐升高，与对照组比较差异有统计学意义（P〈0．05）。免疫荧光双标检测发现L-PGDS主要表达于神经元的树突和星型胶质细胞的胞质胞浆中。结论：L-PGDS在癫痫模型中表达异常升高，可能参与了癫痫的发生发展过程，有望成为临床癫痫诊断的一个分子标志物和新的抗癫痫药物治疗的靶点。
基金funded by the Vice-Rector’s Office at Universidad de Caldas
文摘Background： The purpose of this study was to determine the association of three proteins involved in sperm function on the freezability of porcine semen： the heat shock protein 90 alpha（HSP90a）, the Niemann-Pick disease type C2 protein（NPC2）, and lipocalin-type prostaglandin D synthase（L-PGDS）.Six adult boars（each boar was ejaculated three times, 18 in total） were classified by freezability based on the percentage of functionally competent sperm. The male semen with highest freezability（MHF） and the male semen with lowest freezability（MLF） were centrifuged immediately after collection to separate seminal plasma and spermatozoa to make four possible combinations of these two components and to incubate them for 3 h,adjusting the temperature to 17 °C, to freeze them afterwards. The quantification of proteins was performed in two stages： at zero and at 3 h after incubation of the four combinations.Results： The spermatozoa × incubation time（IT） interaction only had effect（P 〈 0.01） on HSP90 a levels; this protein increased in seminal plasma, after 3 h of incubation, in larger quantity（P 〈 0.05） in combinations with MLF spermatozoa. In relation with the NPC2 protein, two isoforms of 16 and 19 kDa were identified. The 19 kDa isoform was affected（P 〈 0.01） only by the seminal plasma × IT interaction, with superior values（P 〈 0.01） both at zero and three hours of incubation, in the combinations with MHF seminal plasma; and 16 kDa isoform was affected（P 〈 0.01） only by the IT with reduction after 3 h of incubation. The levels of L-PGDS was affected（P 〈 0.01）only by the spermatozoa × IT interaction, which reduced（P 〈 0.01） in combinations with MLF spermatozoa after 3 h of incubation.Conclusions： It is possible to consider that the three proteins evaluated were associated with freezability of boar semen due, especially, to the fact that mixtures with MLF spermatozoa showed greater increase levels of the HSP90 a protein and reduction of L-PGDS in plasma. In addition, the seminal plasma of MHF had higher concentration of the NPC2 of 19 kDa protein, which was reduced by incubating with MHF spermatozoa.
文摘Objectives: Alterations in the prostaglandin-D-system have been found in animal sleep experiments and disorders that present with hypersomnia or sleep disturbances. The recently demonstrated involvement of the leptomeningeal lipocalin- type prostaglandin-Dsynthase (L-PGDS) (β-trace) in human physiological sleep encouraged us to investigate its role in the pathophysiology of narcolepsy. Methods: In a pilot study, serum L-PGDS and melatonin concentrations were assessed in 14 narcoleptic patients during undisturbed sleep and total sleep deprivation, compared with those from 14 healthy controls during undisturbed sleep. Excessive daytime sleepiness was measured by a standardized questionnaire (Epworth slee piness scale, ESS). Results: In narcoleptic patients, markedly increased baseline L-PGDS levels were significantly correlated with the ESS score, but not with the degree of cataplexy. Serum L-PGDS concentrations in patients as well as in controls followed a time-dependent fluctuation with evening increases, highest values during the night and in the morning. Compared with controls, patients exhibited significant/increased amplitude of circulating L-PGDS without any suppression by total sleep deprivation. Conclusion: These findings indicate that the prostaglandin-D-system contributes to the pathophysiology of narcolepsy, e. g. the regulation of excessive daytime sleepiness. Since it has been suggested that L-PGDS is also involved in neurodegenerative disorders, there may be a more specific role of the prostaglandin-D-sys- tem in narcoleptic aetiogenesis. Moreover, its linkage with the immune system as well as with human sleep regulation offers a direct access for investigating both systems.
