安寐汤对失眠大鼠额叶皮质线粒体COXⅠ、ⅣmRNA和蛋白表达的影响
|
摘要
目的探讨安寐汤对失眠大鼠额叶皮质线粒体COXⅠ、ⅣmRNA和蛋白表达的影响。方法取6月龄大鼠进行造模,超活染色观察线粒体功能的完整性,实时荧光定量PCR检测COXⅠ、ⅣmRNA相对表达量,Elisa检测COXⅠ、Ⅳ蛋白表达浓度。结果 (1)与模型组比较,地西泮治疗组和安寐汤低、中、高剂量治疗组均可增加失眠大鼠额叶皮质线粒体COXⅠ、Ⅳ蛋白表达量,且都具有统计学意义(P<0.01)。安寐汤中剂量治疗组对失眠大鼠额叶皮质线粒体COXⅠ、Ⅳ蛋白表达量的增加与低剂量相比有统计学意义(P<0.05),与高剂量之间无统计学意义。(2)安寐汤中剂量治疗组大鼠额叶皮质线粒体COXⅠ、ⅣmRNA表达量明显高于安寐汤低、高剂量治疗组,且具有统计学意义(P<0.05)。结论安寐汤改善大鼠失眠可能与增加额叶皮层线粒体COXⅠ、ⅣmRNA和蛋白表达有关,但剂量-效应关系不明显。
Objective To investigate the effects of Anmei Decoction on mitochondrial COX Ⅰ, Ⅳ mRNA and protein expression in frontal cortex of insomnic rats. Methods Six-month-old rats were used for modeling. The integrity of mitochondrial function was observed by supravital staining. The relative expression of COX Ⅰ, ⅣmRNAs were detected by real-time fluorescent quantitative PCR,and the expression of COXI,Ⅳ proteins were detected by ELISA. Results(1) Compared with the model group, the diazepam treatment group and the low,medium and high doses of Anmei Decoction could increase the expression of COX I,Ⅳ proteins in the frontal cortex of insomnia rats,which were statistically significant(P < 0.01). The effect of the medium dose of Anmei Decoction on the expression of mitochondrial COX I,Ⅳ proteins in frontal cortex of insomnic rats were statistically significant(P < 0.05),and there was no statistical significance between the medium dose and high dose of Anmei Decoction.(2) The expression of mitochondrial COX I,Ⅳ mRNAs in the frontal cortex of the medium dose treatment group of Anmei Decoction were significantly higher than those of the low and high doses treatment groups of Anmei Decoction(P < 0.05). Conclusion Anmei Decoction can improve insomnia, which may be related to the increase of the expression of COXⅠ,Ⅳ mRNAs and proteins of mitochondria in the frontal cortex of the insomnic rats,but the dose-effect relationship is not obvious.
Objective To investigate the effects of Anmei Decoction on mitochondrial COX Ⅰ, Ⅳ mRNA and protein expression in frontal cortex of insomnic rats. Methods Six-month-old rats were used for modeling. The integrity of mitochondrial function was observed by supravital staining. The relative expression of COX Ⅰ, ⅣmRNAs were detected by real-time fluorescent quantitative PCR,and the expression of COXI,Ⅳ proteins were detected by ELISA. Results(1) Compared with the model group, the diazepam treatment group and the low,medium and high doses of Anmei Decoction could increase the expression of COX I,Ⅳ proteins in the frontal cortex of insomnia rats,which were statistically significant(P < 0.01). The effect of the medium dose of Anmei Decoction on the expression of mitochondrial COX I,Ⅳ proteins in frontal cortex of insomnic rats were statistically significant(P < 0.05),and there was no statistical significance between the medium dose and high dose of Anmei Decoction.(2) The expression of mitochondrial COX I,Ⅳ mRNAs in the frontal cortex of the medium dose treatment group of Anmei Decoction were significantly higher than those of the low and high doses treatment groups of Anmei Decoction(P < 0.05). Conclusion Anmei Decoction can improve insomnia, which may be related to the increase of the expression of COXⅠ,Ⅳ mRNAs and proteins of mitochondria in the frontal cortex of the insomnic rats,but the dose-effect relationship is not obvious.
引文
[1]张明芝,武书.苏州市大学生睡眠质量及影响因素的调查研究[J].苏州大学学报(医学版),2009,29(1):99-101.
[2]刘艳骄.安眠药物的替代治疗[J].中国中医基础医学杂志,2006,12(8):628-631,636.
[3]MASSIMINI M,FERRARELLI F,HUBER R,et al.Breakdown of cortical effective connectivity during sleep[J].Science,2005,309(5744):2228-2232.
[4]GERASHCHENKO D,WISOR J P,BURNS D,et al.Identification of a population of sleep-active cerebral cortex neurons[J].Proc Natl Acad Sci USA,2008,105(29):10227-10232.
[5]许晓伍,陈群,马春媚,等.PCPA致大鼠失眠后不同时间点神经元线粒体形态和功能的变化[J].解剖学研究,2014,36(1):34-38.
[6]李宁,唐启盛,侯秀娟,等.广泛性焦虑大鼠前额叶皮质和海马磁共振波谱的研究[J].波谱学杂志,2010,27(2):185-193.
[7]胡佩莹,林伟山,李莉,等.大通犊牦牛组织线粒体COX酶活性与亚基COXⅠ、ⅡmRNA表达的相关性研究[J].中国兽医学报,2015,35(8):1468-1471.
[8]许晓伍,陈群,郝木峰,等.夜交藤提取物对失眠大鼠额叶皮层神经元线粒体结构和功能的影响[J].广州中医药大学学报,2013,30(6):872-875.
[9]许晓伍,葛振英,石纯,等.植物雌激素对去卵巢大鼠海马神经元线粒体超微结构的保护作用[J].神经解剖学杂志,2006,22(3):351-355.
[10]刘琼丽,易继涛,李玉山,等.夜交藤提取物对小鼠中枢神经系统作用的实验研究[J].临床和实验医学杂志,2014,13(17):1404-1407.
[11]武静,王慧,史琴,等.PCPA失眠大鼠不同时间皮质前额叶ATP水平变化及酸枣仁汤的干预作用[J].重庆医学,2017,46(4):439-441.
[12]陈群,徐志伟.安寐汤治疗失眠126例疗效观察[J].新中医,2000,32(6):18-19.
[13]刁远明,陈群.安寐汤对失眠大鼠脑组织多巴胺含量的影响[J].中国医药导报,2011,8(17):105-107.
[14]左文彪,臧印竹,王慧.酸枣仁汤对PCPA致失眠大鼠大脑皮质代谢型谷氨酸受体mGluR1、mGluR2的影响[J].亚太传统医药,2018,14(3):4-7.
[15]王亚茹,于世家.于世家教授自拟安眠方对PCPA诱导失眠大鼠脑内递质及GABA的影响[J].时珍国医国药,2018,29(4):852-855.
[16]吴建丽,韩璐,刘旭东,等.不同频率电项针对PCPA致失眠大鼠睡眠时相及血清ACTH、CORT含量的影响[J].吉林中医药,2018,38(2):191-195.
[17]李峰杰,何萍,赵乐,等.酸枣仁、石菖蒲、夜交藤、百合、郁金5味中药对对氯苯丙氨酸致失眠模型大鼠睡眠作用的影响[J].中国药业,2017,26(6):1-4.
[18]黄杰聪,谢炜,邓宁,等.佛手宁神方改善睡眠作用的机制[J].南方医科大学学报,2017,37(8):1116-1120.
[19]吴巧敏,赵艺初,韩艺凡,等.生酸枣仁、炒酸枣仁、酸枣果肉对PCPA失眠模型大鼠学习记忆能力影响的对比研究[J].中医药导报,2016,22(5):72-75.
[20]顾思臻,窦丹波.中医中药对PCPA失眠大鼠模型HPA轴相关单胺类神经递质及激素影响的研究进展[J].上海中医药大学学报,2015,29(1):83-87.
[21]王雅丽,黄俊山.神经递质与睡眠觉醒及学习记忆的关系研究[J].医学综述,2011,17(1):15-18.
[22]高家荣,季文博,姜辉,等.酸枣仁-五味子药对醇水双提物对PCPA致失眠大鼠氨基酸类神经递质的影响[J].中药材,2013,36(10):1635-1639.
[23]POLSTER B M,FISKUM C L.Mitochondrial mechanisms of neural cell apoptosis[J].J Neurochem,2004,90(6):1281-1289.
[24]成令忠.组织学(第二版)[M].北京:人民卫生出版社,1993:95.
[25]郭婉华.基础分子细胞生物学[M].广东:广东科技出版社,2000:140-142.
[26]KEILIN,D.Cytochrome and respiratory enzymes[J].Proceedings of the Royal Society of London,1929,104(730):206-252.
[27]王苏华,张薇,邢光伟,等.细胞色素C氧化酶活力测定方法概况[J].毒理学杂志,2010,24(5):412-414.
[28]孙飞,周强军,孙吉,等.线粒体呼吸链膜蛋白复合体的结构[J].生命科学,2008,20(4):566-579.
[29]赵晴,杜建时,曲淑彦,等.血管性痴呆大鼠海马线粒体COX活性及基因的表达变化的研究[J].中国实验诊断学,2009,13(9):1144-1149.
[30]柏干荣,陆松敏.线粒体DNA编码细胞色素氧化酶亚基基因的进展[J].国外医学(分子生物学分册),2003,25(6):354-358.
[31]周秀敏,杨永江,毕英杰,等.基于线粒体?COXⅠ基因变异探讨国内外猪种的遗传多样性及系统进化研究[J].浙江农业学报,2017,29(8):1271-1280.
[32]立彦,王自正.线粒体DNA的研究现状[J].国际检验医学杂志,2006,27(7):636-638.
[2]刘艳骄.安眠药物的替代治疗[J].中国中医基础医学杂志,2006,12(8):628-631,636.
[3]MASSIMINI M,FERRARELLI F,HUBER R,et al.Breakdown of cortical effective connectivity during sleep[J].Science,2005,309(5744):2228-2232.
[4]GERASHCHENKO D,WISOR J P,BURNS D,et al.Identification of a population of sleep-active cerebral cortex neurons[J].Proc Natl Acad Sci USA,2008,105(29):10227-10232.
[5]许晓伍,陈群,马春媚,等.PCPA致大鼠失眠后不同时间点神经元线粒体形态和功能的变化[J].解剖学研究,2014,36(1):34-38.
[6]李宁,唐启盛,侯秀娟,等.广泛性焦虑大鼠前额叶皮质和海马磁共振波谱的研究[J].波谱学杂志,2010,27(2):185-193.
[7]胡佩莹,林伟山,李莉,等.大通犊牦牛组织线粒体COX酶活性与亚基COXⅠ、ⅡmRNA表达的相关性研究[J].中国兽医学报,2015,35(8):1468-1471.
[8]许晓伍,陈群,郝木峰,等.夜交藤提取物对失眠大鼠额叶皮层神经元线粒体结构和功能的影响[J].广州中医药大学学报,2013,30(6):872-875.
[9]许晓伍,葛振英,石纯,等.植物雌激素对去卵巢大鼠海马神经元线粒体超微结构的保护作用[J].神经解剖学杂志,2006,22(3):351-355.
[10]刘琼丽,易继涛,李玉山,等.夜交藤提取物对小鼠中枢神经系统作用的实验研究[J].临床和实验医学杂志,2014,13(17):1404-1407.
[11]武静,王慧,史琴,等.PCPA失眠大鼠不同时间皮质前额叶ATP水平变化及酸枣仁汤的干预作用[J].重庆医学,2017,46(4):439-441.
[12]陈群,徐志伟.安寐汤治疗失眠126例疗效观察[J].新中医,2000,32(6):18-19.
[13]刁远明,陈群.安寐汤对失眠大鼠脑组织多巴胺含量的影响[J].中国医药导报,2011,8(17):105-107.
[14]左文彪,臧印竹,王慧.酸枣仁汤对PCPA致失眠大鼠大脑皮质代谢型谷氨酸受体mGluR1、mGluR2的影响[J].亚太传统医药,2018,14(3):4-7.
[15]王亚茹,于世家.于世家教授自拟安眠方对PCPA诱导失眠大鼠脑内递质及GABA的影响[J].时珍国医国药,2018,29(4):852-855.
[16]吴建丽,韩璐,刘旭东,等.不同频率电项针对PCPA致失眠大鼠睡眠时相及血清ACTH、CORT含量的影响[J].吉林中医药,2018,38(2):191-195.
[17]李峰杰,何萍,赵乐,等.酸枣仁、石菖蒲、夜交藤、百合、郁金5味中药对对氯苯丙氨酸致失眠模型大鼠睡眠作用的影响[J].中国药业,2017,26(6):1-4.
[18]黄杰聪,谢炜,邓宁,等.佛手宁神方改善睡眠作用的机制[J].南方医科大学学报,2017,37(8):1116-1120.
[19]吴巧敏,赵艺初,韩艺凡,等.生酸枣仁、炒酸枣仁、酸枣果肉对PCPA失眠模型大鼠学习记忆能力影响的对比研究[J].中医药导报,2016,22(5):72-75.
[20]顾思臻,窦丹波.中医中药对PCPA失眠大鼠模型HPA轴相关单胺类神经递质及激素影响的研究进展[J].上海中医药大学学报,2015,29(1):83-87.
[21]王雅丽,黄俊山.神经递质与睡眠觉醒及学习记忆的关系研究[J].医学综述,2011,17(1):15-18.
[22]高家荣,季文博,姜辉,等.酸枣仁-五味子药对醇水双提物对PCPA致失眠大鼠氨基酸类神经递质的影响[J].中药材,2013,36(10):1635-1639.
[23]POLSTER B M,FISKUM C L.Mitochondrial mechanisms of neural cell apoptosis[J].J Neurochem,2004,90(6):1281-1289.
[24]成令忠.组织学(第二版)[M].北京:人民卫生出版社,1993:95.
[25]郭婉华.基础分子细胞生物学[M].广东:广东科技出版社,2000:140-142.
[26]KEILIN,D.Cytochrome and respiratory enzymes[J].Proceedings of the Royal Society of London,1929,104(730):206-252.
[27]王苏华,张薇,邢光伟,等.细胞色素C氧化酶活力测定方法概况[J].毒理学杂志,2010,24(5):412-414.
[28]孙飞,周强军,孙吉,等.线粒体呼吸链膜蛋白复合体的结构[J].生命科学,2008,20(4):566-579.
[29]赵晴,杜建时,曲淑彦,等.血管性痴呆大鼠海马线粒体COX活性及基因的表达变化的研究[J].中国实验诊断学,2009,13(9):1144-1149.
[30]柏干荣,陆松敏.线粒体DNA编码细胞色素氧化酶亚基基因的进展[J].国外医学(分子生物学分册),2003,25(6):354-358.
[31]周秀敏,杨永江,毕英杰,等.基于线粒体?COXⅠ基因变异探讨国内外猪种的遗传多样性及系统进化研究[J].浙江农业学报,2017,29(8):1271-1280.
[32]立彦,王自正.线粒体DNA的研究现状[J].国际检验医学杂志,2006,27(7):636-638.