生慧汤对慢性睡眠剥夺小鼠学习记忆及海马IL-6,TNF-α,COX-2基因表达的影响
|
摘要
目的:通过观察生慧汤对慢性睡眠剥夺小鼠学习记忆及海马白细胞介素-6(interleukin-6,IL-6),肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α),环氧化酶-2(cyclooxygenase-2,COX-2) mRNA表达的影响,探索生慧汤改善学习记忆能力的可能机制。方法:将实验小鼠随机分为睡眠剥夺模型组,正常组,褪黑素组(7. 8×10-4g·kg~(-1)·d~(-1)),生慧汤高、中、低剂量组(54,27,13. 5 g·kg~(-1)·d~(-1)),采用多平台水环境法建立小鼠慢性睡眠剥夺模型,睡眠剥夺28 d并灌胃给药。采用Morris水迷宫检测各组小鼠学习记忆能力;采用实时荧光定量聚合酶链式反应(Real-time PCR)法检测各组小鼠海马IL-6,TNF-α,COX-2mRNA表达量。结果:Morris水迷宫试验结果显示,与正常组比较,模型组找到平台消耗时间与游泳总路程明显延长(P <0. 01),穿越平台次数与目标象限时间明显减少(P <0. 01),第1次抵原平台时间明显增加(P <0. 01);与模型组比较,各给药组找到平台消耗时间与游泳总路程明显减少(P <0. 05,P <0. 01),穿越平台次数与目标象限时间明显增加(P <0. 05,P <0. 01),第1次抵原平台时间明显减少(P <0. 05,P <0. 01)。Real-time PCR检测结果显示,与正常组比较,模型组小鼠IL-6,TNF-α,COX-2 mRNA表达上调(P <0. 01);与模型组比较,各给药组IL-6,TNF-α,COX-2 mRNA表达下调(P <0. 05,P <0. 01),生慧汤低剂量组TNF-α,COX-2 mRNA表达无显著性差异。结论:生慧汤可能通过降低海马中IL-6,TNF-α,COX-2mRNA的表达来改善小鼠学习记忆能力。
Objective: To observe the effect of Shenghuitang on learning and memory and expressions of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α) and cyclooxygenase-2(COX-2) in hippocampus of chronic sleep deprived mice,in order to explore the possible mechanism of Shenghuitang in improving learning and memory ability. Method: Mice were randomly divided into sleep deprivation group,blank group,melatonin group(7. 8 × 10-4 g·kg~(-1)·d~(-1)),high,middle and low-dose Shenghuitang groups(54,27,13. 5 g·kg~(-1)·d~(-1)). The model of chronic sleep deprivation in mice was established using the " multi-platform water environment method".28 d sleep deprivation and intragastric administration were provided. Morris water maze was used to detect the learning and memory ability of mice in each group. Real time-PCR was used to detect mRNA expressions of IL-6,TNF-α and COX-2 in the hippocampus of each group. Result: The results of Morris water maze test showed that compared with the blank group,the total time spent on finding the platform and the total swimming distance of the model group were significantly prolonged(P < 0. 01),while the number of crossing platforms and the target quadrant were significantly reduced(P < 0. 01). The time for the original platform was significantly increased(P < 0. 01). Compared with the model group,the total time spent on finding the platform and the total swimming distance decreased significantly in each drug-administered group(P < 0. 05,P < 0. 01) reduced,whereas the number of times for crossing the platform and the target quadrant increased significantly(P < 0. 05,P < 0. 01).The time for the first arrival of the original platform was significantly reduced(P < 0. 05,P < 0. 01). The results of RT-PCR showed that mRNA expressions of IL-6,TNF-α,and COX-2 were increased in the model group compared with the blank group. Compared with the model group,mRNA expressions of IL-6,TNF-α,and COX-2 were decreased in the treated group. COX-2 mRNA expression was down-regulated. Conclusion: Shenghuitang may improve the learning and memory ability of mice by decreasing mRNA expressions of IL-6,TNF-α and COX-2 in hippocampus.
Objective: To observe the effect of Shenghuitang on learning and memory and expressions of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α) and cyclooxygenase-2(COX-2) in hippocampus of chronic sleep deprived mice,in order to explore the possible mechanism of Shenghuitang in improving learning and memory ability. Method: Mice were randomly divided into sleep deprivation group,blank group,melatonin group(7. 8 × 10-4 g·kg~(-1)·d~(-1)),high,middle and low-dose Shenghuitang groups(54,27,13. 5 g·kg~(-1)·d~(-1)). The model of chronic sleep deprivation in mice was established using the " multi-platform water environment method".28 d sleep deprivation and intragastric administration were provided. Morris water maze was used to detect the learning and memory ability of mice in each group. Real time-PCR was used to detect mRNA expressions of IL-6,TNF-α and COX-2 in the hippocampus of each group. Result: The results of Morris water maze test showed that compared with the blank group,the total time spent on finding the platform and the total swimming distance of the model group were significantly prolonged(P < 0. 01),while the number of crossing platforms and the target quadrant were significantly reduced(P < 0. 01). The time for the original platform was significantly increased(P < 0. 01). Compared with the model group,the total time spent on finding the platform and the total swimming distance decreased significantly in each drug-administered group(P < 0. 05,P < 0. 01) reduced,whereas the number of times for crossing the platform and the target quadrant increased significantly(P < 0. 05,P < 0. 01).The time for the first arrival of the original platform was significantly reduced(P < 0. 05,P < 0. 01). The results of RT-PCR showed that mRNA expressions of IL-6,TNF-α,and COX-2 were increased in the model group compared with the blank group. Compared with the model group,mRNA expressions of IL-6,TNF-α,and COX-2 were decreased in the treated group. COX-2 mRNA expression was down-regulated. Conclusion: Shenghuitang may improve the learning and memory ability of mice by decreasing mRNA expressions of IL-6,TNF-α and COX-2 in hippocampus.
引文
[1]徐健,颜崇淮,沈晓明.睡眠剥夺损害学习记忆能力的研究[J].中华预防医学杂志,2004,38(2):134-137.
[2]赵忠新,张红菊,黄流清.失眠的治疗药物及其使用方法研究进展[J].中国新药与临床杂志,2007,26(11):851-856.
[3]陆瑾.调和营卫法针刺治疗失眠症临床观察[J].上海针灸杂志,2008,27(2):6-7.
[4]张如意,王平,张舜波,等.褪黑素治疗睡眠障碍的作用机制探讨[J].中华中医药学刊,2018,36(2):308-310.
[5]丁莉,游秋云,王平.生慧汤对APP/PS1双转基因痴呆小鼠自主活动昼夜节律的影响及对下丘脑生物钟基因的调节[J].时珍国医国药,2017,28(2):301-304.
[6]周敏,白兰,周丽娜,等.生慧汤对学习记忆障碍模型小鼠的影响[J].中医药信息,2009,26(6):54-55.
[7]程玥,陈淑娴,张雪,等.生慧汤对糖尿病脑病模型大鼠认知障碍及神经病理改变的影响[J].中成药,2015,37(12):2579-2584.
[8]崔开宇,王平,游秋云.益智仁挥发油对大鼠快动眼睡眠剥夺恢复后脑组织氨基酸类神经递质含量及其受体表达的影响[J].中国实验方剂学杂志,2013,19(4):223-227.
[9]张舜波,王平,田代志,等.酸枣仁总皂苷对失眠老年大鼠脑氨基酸类神经递质及受体表达的影响[J].中国实验方剂学杂志,2014,20(4):124-127.
[10]马哲,王平,游秋云.睡眠及睡眠剥夺与学习记忆的相关性探讨[J].中华中医药杂志,2014,29(4):995-997.
[11] CHANG H M,LIAO W C,Sheu J N,et al. Sleep deprivation impairs Ca2+expression in the hippocampus:ionic imaging analysis for cognitive deficiency with TOF-SIMS[J]. Microsc Microanal,2012,18(3):425-435.
[12] Graves L,Pack A,Abel T. Sleep and memory:a molecular perspective[J]. Trends in Neurosciences,2001,24(4):237-243.
[13] Everson C A,Laatsch C D,Hogg N. Antioxidant defense responses to sleep loss and sleep recovery[J]. Am J Physiol Regul Integr Comp Physiol, 2005, 288(2):R374.
[14] Baker F C,Shah S,Stewart D,et al. Interleukin 1beta enhances non-rapid eye movement sleep and increases cFos protein expression in the median preoptic nucleus of the hypothalamus[J]. Am J Physiol Regul Integr Comp Physiol,2005,288(4):R998.
[15]尚琦松,黄擘,盛文辉,等.炎性因子TNF-α及IL-18与椎间盘退变的相关性研究[J].中国矫形外科杂志,2009,17(5):385-387.
[16] WONG Y F,ZHOU H,WANG J R,et al. Antiinflammatory and analgesic effects and molecular mechanisms of JCICM-6,a purified extract derived from an anti-arthritic Chinese herbal formula[J].Phytomedicine,2008,15(6):416-426.
[17]曲道炜,朱辉,杜斌,等.桂芍知母汤及加味对佐剂性关节炎大鼠炎性因子及COX-2信号通路表达的影响[J].中华中医药杂志,2015,30(5):1719-1722.
[2]赵忠新,张红菊,黄流清.失眠的治疗药物及其使用方法研究进展[J].中国新药与临床杂志,2007,26(11):851-856.
[3]陆瑾.调和营卫法针刺治疗失眠症临床观察[J].上海针灸杂志,2008,27(2):6-7.
[4]张如意,王平,张舜波,等.褪黑素治疗睡眠障碍的作用机制探讨[J].中华中医药学刊,2018,36(2):308-310.
[5]丁莉,游秋云,王平.生慧汤对APP/PS1双转基因痴呆小鼠自主活动昼夜节律的影响及对下丘脑生物钟基因的调节[J].时珍国医国药,2017,28(2):301-304.
[6]周敏,白兰,周丽娜,等.生慧汤对学习记忆障碍模型小鼠的影响[J].中医药信息,2009,26(6):54-55.
[7]程玥,陈淑娴,张雪,等.生慧汤对糖尿病脑病模型大鼠认知障碍及神经病理改变的影响[J].中成药,2015,37(12):2579-2584.
[8]崔开宇,王平,游秋云.益智仁挥发油对大鼠快动眼睡眠剥夺恢复后脑组织氨基酸类神经递质含量及其受体表达的影响[J].中国实验方剂学杂志,2013,19(4):223-227.
[9]张舜波,王平,田代志,等.酸枣仁总皂苷对失眠老年大鼠脑氨基酸类神经递质及受体表达的影响[J].中国实验方剂学杂志,2014,20(4):124-127.
[10]马哲,王平,游秋云.睡眠及睡眠剥夺与学习记忆的相关性探讨[J].中华中医药杂志,2014,29(4):995-997.
[11] CHANG H M,LIAO W C,Sheu J N,et al. Sleep deprivation impairs Ca2+expression in the hippocampus:ionic imaging analysis for cognitive deficiency with TOF-SIMS[J]. Microsc Microanal,2012,18(3):425-435.
[12] Graves L,Pack A,Abel T. Sleep and memory:a molecular perspective[J]. Trends in Neurosciences,2001,24(4):237-243.
[13] Everson C A,Laatsch C D,Hogg N. Antioxidant defense responses to sleep loss and sleep recovery[J]. Am J Physiol Regul Integr Comp Physiol, 2005, 288(2):R374.
[14] Baker F C,Shah S,Stewart D,et al. Interleukin 1beta enhances non-rapid eye movement sleep and increases cFos protein expression in the median preoptic nucleus of the hypothalamus[J]. Am J Physiol Regul Integr Comp Physiol,2005,288(4):R998.
[15]尚琦松,黄擘,盛文辉,等.炎性因子TNF-α及IL-18与椎间盘退变的相关性研究[J].中国矫形外科杂志,2009,17(5):385-387.
[16] WONG Y F,ZHOU H,WANG J R,et al. Antiinflammatory and analgesic effects and molecular mechanisms of JCICM-6,a purified extract derived from an anti-arthritic Chinese herbal formula[J].Phytomedicine,2008,15(6):416-426.
[17]曲道炜,朱辉,杜斌,等.桂芍知母汤及加味对佐剂性关节炎大鼠炎性因子及COX-2信号通路表达的影响[J].中华中医药杂志,2015,30(5):1719-1722.