孕酮对小鼠放射性脑损伤的神经保护作用及分子机制
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摘要
目的探讨孕酮对小鼠放射性脑损伤的神经保护作用及其可能分子机制。方法 144只小鼠分为溶剂对照组、单纯照射组和孕酮组,采用Morris水迷宫实验评价小鼠空间学习和记忆能力;干/湿法测定脑含水量;ELISA法检测脑脊液三磷酸腺苷(ATP)、环氧合酶-2(COX-2)、肿瘤坏死因子-α(TNF-α)和白介素-6(IL-6)的含量;Western blot法检测脑组织P2X7受体蛋白表达水平。结果单纯照射组小鼠第4~7天平均潜伏期时间和寻找平台的路径总长均明显大于溶剂对照组,孕酮组明显短于单纯照射组(P=0.000 1);单纯照射组脑含水量和细胞凋亡率明显高于溶剂对照组,孕酮组明显低于单纯照射组(P=0.000 0);单纯照射组脑脊液ATP、COX-2、TNF-α和IL-6的含量明显高于溶剂对照组,孕酮组明显低于单纯照射组(P=0.010 9、0.004 6、0.007 8、0.001 4);单纯照射组小鼠脑组织P2X7受体蛋白的表达高于溶剂对照组,而孕酮组P2X7受体蛋白的表达水平明显低于单纯照射组。结论孕酮可能通过抑制ATP介导的P2X7R活化和炎症介质的释放,进而减轻脑水肿和减少细胞凋亡,起到保护放射性脑损伤所致的神经损害作用。
Objective To investigate the neuroprotective effects of progesterone on radiation-induced brain injury in mice and its possible molecular mechanism. Methods 144 mice were divided into control group,radiation group and progesterone group. The spatial learning and memory ability of mice were evaluated by Morris water maze test;The water content in brain of mice were measured by dry/wet method; The levels of adenosine triphosphate( ATP),cyclooxygenase-2( COX-2),tumor necrosis factor alpha( TNF-α) and interleukin-6( IL-6) in cerebrospinal fluid were determined by ELISA. The expression of P2 X7 receptor protein in brain tissue was detected by Western blot. Results The average latency time and the length of path of finding platform on fourth to seventh day after training in radiation group were significantly longer than the control group,progesterone group was significantly shorter than that in radiation group( P = 0. 000 1). The brain water content and the rate of apoptosis in radiation group were significantly higher than the control group,progesterone group was significantly lower than that of the radiation group( P = 0. 000 0). The content of ATP,COX-2,TNF-α and IL-6 in cerebrospinal fluid of radiation group were significantly higher than the control group,progesterone group was significantly lower than that in the radiation group( P = 0. 010 9,0. 004 6,0. 007 8,0. 001 4). The expression of P2 X7 receptor protein in brain tissue of radiation group increased significantly compared with the control group,while progesterone group was significantly lower than that of the radiation group. Conclusion Progesterone may inhibit neuropathy caused by radiation-induced brain injury by inhibiting ATP-mediated activation of P2 X7 R and release of inflammatory mediators,thereby reducing brain edema and reducing apoptosis.
Objective To investigate the neuroprotective effects of progesterone on radiation-induced brain injury in mice and its possible molecular mechanism. Methods 144 mice were divided into control group,radiation group and progesterone group. The spatial learning and memory ability of mice were evaluated by Morris water maze test;The water content in brain of mice were measured by dry/wet method; The levels of adenosine triphosphate( ATP),cyclooxygenase-2( COX-2),tumor necrosis factor alpha( TNF-α) and interleukin-6( IL-6) in cerebrospinal fluid were determined by ELISA. The expression of P2 X7 receptor protein in brain tissue was detected by Western blot. Results The average latency time and the length of path of finding platform on fourth to seventh day after training in radiation group were significantly longer than the control group,progesterone group was significantly shorter than that in radiation group( P = 0. 000 1). The brain water content and the rate of apoptosis in radiation group were significantly higher than the control group,progesterone group was significantly lower than that of the radiation group( P = 0. 000 0). The content of ATP,COX-2,TNF-α and IL-6 in cerebrospinal fluid of radiation group were significantly higher than the control group,progesterone group was significantly lower than that in the radiation group( P = 0. 010 9,0. 004 6,0. 007 8,0. 001 4). The expression of P2 X7 receptor protein in brain tissue of radiation group increased significantly compared with the control group,while progesterone group was significantly lower than that of the radiation group. Conclusion Progesterone may inhibit neuropathy caused by radiation-induced brain injury by inhibiting ATP-mediated activation of P2 X7 R and release of inflammatory mediators,thereby reducing brain edema and reducing apoptosis.
引文
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[12]Oh S B,Park H R,Jang Y J,et al.Baicalein attenuates impaired hippocampal neurogenesis and the neurocognitive deficits induced byγ-ray radiation[J].Br J Pharmaeol,2013,168(2):421-31.
[13]雷贤明,曹云涛.孕酮神经保护作用研究进展[J].海南医学,2014,25(4):544-6.
[14]Carmo M R,Menezes A P,Nunes A C,et al.The P2X7receptor antagonist brilliant blue G attenuates contralateral rotations in a rat model of Parkinsonism through a combined control of synaptotoxicity,neurotoxicity and gliosis[J].Neuropharmacology,2014,81:142-52.
[15]Sperlágh B,Illes P.P2X7receptor:an emerging target in central nervous system diseases[J].Trends Pharmacol Sci,2014,35(10):537-47.
[2]李新娟,李爽,李东亮.孕酮对缺氧缺血性脑损伤新生大鼠血脑屏障通透性及脑水肿的影响[J].中国药理学通报,2011,27(3):364-7.
[3]李新娟,何瑞芳,李爽,等.孕酮对全脑缺血/再灌注损伤大鼠学习记忆及海马P2X_7受体表达的影响[J].中国应用生理学杂志,2012,28(5):472-5.
[4]王丽雁,蔡文琴.P2X7受体在神经系统表达及功能的研究进展[J].解剖学杂志,2007,30(6):813-6.
[5]Xu P,Xu Y,Hu B,et al.Extracellular ATP enhances radiationinduced brain injury through microglial activation and paracrine signaling via P2X7receptor[J].Brain Behav Immun,2015,50:87-100.
[6]Tang Y,Rong X,Hu W,et al.Effect of edaravone on radiationinduced brain necrosis in patients with nasopharyngeal carcinoma after radiotherapy:a randomized controlled trial[J].J Neurooncol,2014,120(2):441-7.
[7]Panagiotakos G,Alshamy G,Chan B,et al.Long-term impact of radiation on the stem cell and oligodendrocyte precursors in the brain[J].PLo S One,2007,2(7):e588.
[8]Peng Y,Lu K,Li Z,et al.Blockade of Kv1.3 channels ameliorates radiation-induced brain injury[J].Neuro Oncol,2014,16(4):528-39.
[9]Dong X,Luo M,Huang G,et al.Relationship between irradiationinduced neuro-inflammatory environments and impaired cognitive function in the developing brain of mice[J].Int J Radiat Biol,2015,91(3):224-39.
[10]张海博,梁海乾,李卫民,等.放射性脑损伤的研究现状[J].山东医药,2014,54(26):95-7.
[11]郑慧芬,涂或.硫酸镁对急性放射性脑损伤后神经细胞凋亡的影响[J].现代中西医结合杂志,2009,18(4):364-7.
[12]Oh S B,Park H R,Jang Y J,et al.Baicalein attenuates impaired hippocampal neurogenesis and the neurocognitive deficits induced byγ-ray radiation[J].Br J Pharmaeol,2013,168(2):421-31.
[13]雷贤明,曹云涛.孕酮神经保护作用研究进展[J].海南医学,2014,25(4):544-6.
[14]Carmo M R,Menezes A P,Nunes A C,et al.The P2X7receptor antagonist brilliant blue G attenuates contralateral rotations in a rat model of Parkinsonism through a combined control of synaptotoxicity,neurotoxicity and gliosis[J].Neuropharmacology,2014,81:142-52.
[15]Sperlágh B,Illes P.P2X7receptor:an emerging target in central nervous system diseases[J].Trends Pharmacol Sci,2014,35(10):537-47.