HSE患儿及小鼠脑脊液GABA表达及其受体激动剂对炎症因子的影响
|
摘要
目的探讨单纯疱疹病毒性脑炎(HSE)患儿及小鼠脑脊液γ-氨基丁酸(GABA)的表达水平;检测GABA受体激动剂干预前后,HSE小鼠脑脊液内炎性因子S100B、TNF-α、IL-2的表达情况。方法选取2014年10月～2016年1月于我院就诊并符合HSE诊断标准的患儿10例为HSE组,同期选取在我院就诊有腰穿指征但临床排除中枢神经系统感染的儿童10名为对照组;在4周龄SPF级雄性C57BL/6小鼠50只中随机选取10只分为空白对照组(5只)和生理盐水组(5只),剩余40只用于HSV-1型(HSE)模型的构建,成功构建25只,选取5只为HSE-1组,剩余20只分为HSE-2组、HSE-2+生理盐水组、HSE-2+受体激动剂A组、HSE-2+受体激动剂B组,并经GABA受体激动剂干预。ELISA或RT-PCR法检测不同组中GABA、S100B、TNF-α、IL-2的表达水平。结果 HSE组患儿脑脊液常规检查(CSF)中GABA水平明显低于正常组(P<0.05);HSE小鼠中HSE-1组的GABA含量明显低于空白对照组(P<0.01),空白对照组与生理盐水组组间比较差异无统计学意义(P>0.05);HSE各组小鼠脑组织中S100B、TNF-α、IL-2的mRNA表达明显高于空白对照组(P<0.05),经GABA受体激动剂干预后,HSE-2+受体激动剂A组S100B、TNF-α、IL-2的表达量与HSE-2组相比明显降低(P=0.0028),而HSE-2+受体激动剂B组表达量与HSE-2组相比差异无统计学意义(P>0.05);HSE-2组与HSE-2+生理盐水组S100B、TNF-α、IL-2含量相比差异无统计学意义(P>0.05)。结论 HSE患儿及小鼠脑组织中GABA含量均下降,HSE小鼠脑组织中S100B、TNF-α、IL-2 mRNA含量升高。GABA受体激动剂可以降低HSV-1型HSE小鼠颅内的炎性因子。
Objective To investigate GABA expression in HSE children and HSE mice. The expression levels of S100 B, TNF-α and IL-2 in HSE mice were detected before and after GABA receptor agonist used. Methods The HSE children and the control group were collected from 2014 to 2016. The levels of GABA in cerebrospinal fluid(CSF) were detected by ELISA. 40 mice were selected from 50 of 4 year old male C57 BL/6 mice for hsv-1(HSE) mouse model. 25 mice model were successfully constructed. The expression levels of GABA or S100 B, TNF-α and IL-2 in different groups(control group, normal saline group, HSE-1 group, HSE-2 group HSE-2+normal saline group, HSE-2+ receptor agonist A group, HSE-2+ receptor agonist B group) were detected by ELISA or RT-PCR.Results The GABA level of CSF in HSE was significantly lower than that in normal children. Compared with the blank control group and the sham operation group, the GABA of CSF in HSV-1 mice was significantly decreased, and the mRNA expression of S100 B, TNF-α and IL-2 in brain tissue was significantly increased. The mRNA expression of S100 B, TNF-α and IL-2 was significantly decreased after muscimol intervention. Conclusion The GABA levels of CSF were significantly decreased in HSE children or mice. However, the levels of the inflammatory factors including S100 B, TNF-α and IL-2 in HSE mice were significantly increased. GABA receptor agonists can reduce the intracranial inflammatory factors of HSE mice.
Objective To investigate GABA expression in HSE children and HSE mice. The expression levels of S100 B, TNF-α and IL-2 in HSE mice were detected before and after GABA receptor agonist used. Methods The HSE children and the control group were collected from 2014 to 2016. The levels of GABA in cerebrospinal fluid(CSF) were detected by ELISA. 40 mice were selected from 50 of 4 year old male C57 BL/6 mice for hsv-1(HSE) mouse model. 25 mice model were successfully constructed. The expression levels of GABA or S100 B, TNF-α and IL-2 in different groups(control group, normal saline group, HSE-1 group, HSE-2 group HSE-2+normal saline group, HSE-2+ receptor agonist A group, HSE-2+ receptor agonist B group) were detected by ELISA or RT-PCR.Results The GABA level of CSF in HSE was significantly lower than that in normal children. Compared with the blank control group and the sham operation group, the GABA of CSF in HSV-1 mice was significantly decreased, and the mRNA expression of S100 B, TNF-α and IL-2 in brain tissue was significantly increased. The mRNA expression of S100 B, TNF-α and IL-2 was significantly decreased after muscimol intervention. Conclusion The GABA levels of CSF were significantly decreased in HSE children or mice. However, the levels of the inflammatory factors including S100 B, TNF-α and IL-2 in HSE mice were significantly increased. GABA receptor agonists can reduce the intracranial inflammatory factors of HSE mice.
引文
[1] 陈爱华,周辉.儿童单纯疱疹病毒性脑炎临床特点与预后分析[J].中国实用神经疾病杂志,2015,18(04):49-50.
[2] Kennedy P G.Herpes simplex virus and the nervous system[J].Postgrad Medical Journal,1984,60(702):253-259.
[3] Nahmias A J,Lee F K,Beckman-Nahmias S.Seroepidemiological and sociological patterns of herpes simplex virus infection in the world[J].Scand J Infect Dis Suppl,1990,69(69):19-36.
[4] 周渊峰,周水珍,曹凌锋,等.儿童单纯疱疹病毒性脑炎 20 例临床分析及随访研究[J].中国实用儿科杂志,2011,26(7):527-530.
[5] Lellouch-Tubiana A,Fohlen M,Robain O,et al.Immunocytochemical characterization of long-term persistent immune activation in human brain after herpes simplex encephalitis[J].Neuropathol Appl Neurobiol,2000,26(3):285-294.
[6] Kapur N,Barker S,Burrows E H,et al.Herpes simplex encephalitis:long term magnetic resonance imaging and neuropsychological profile[J].J Neurol Neurosurg Psychiatry,1994,57(11):1334-1342.
[7] Costa E,Auta J,Grayson D R,et al.GABAA receptors and benzodiazepines:a role for dendritic resident subunit mRNAs[J].Neuropharmacology,2002,43(6):925-937.
[8] Bhat R,Axtell R,Mitra A,et al.Inhibitory role for GABA in autoimmune inflammation[J].Proc Natl Acad Sci U S A,2010,107(6):2580-2585.
[9] 衣曼,魏克伦,王雪峰,等.小鼠单纯疱疹病毒性脑炎模型的建立[J].小儿急救医学,2000,7(1):26-27.
[10] Satpathy G,Mishra A K,Tandon R,et al.Evaluation of tear samples for Herpes Simplex Virus 1 (HSV) detection in suspected cases of viral keratitis using PCR assay and conventional laboratory diagnostic tools[J].Br J Ophthalmol,2011,95(3):415-418.
[11] Ulett G C,Ketheesan N,Hirst R G.Cytokine gene expression in innately susceptible BALB/c mice and relatively resistant C57BL/6 mice during infection with virulent Burkholderia pseudomallei[J].Infect Immun,2000,68(4):2034-2042.
[12] Dirig D M,Yaksh T L.Intrathecal baclofen and muscimol,but not midazolam,are antinociceptive using the rat-formalin model[J].J Pharmacol Exp Ther,1995,275(1):219-227.
[13] Huse M,Lillemeier B F,Kuhns M S,et al.T cells use two directionally distinct pathways for cytokine secretion[J].Nat Immunol,2006,7(3):247-255.
[14] Hukkanen V,Broberg E,Salmi A,et al.Cytokines in experimental herpes simplex virus infection[J].Int Rev Immunol,2002,21(4-5):355-371.
[15] Katan M,Moon Y P,Paik M C,et al.Infectious burden and cognitive function:the Northern Manhattan Study[J].Neurology,2013,80(13):1209-1215.
[16] Gajcy K,Lochynski S,Librowski T.A role of GABA analogues in the treatment of neurological diseases[J].Curr Med Chem,2010,17(22):2338-2347.
[17] Goren M Z,Kucukibrahimoglu E,Berkman K,et al.Fluoxetine partly exerts its actions through GABA:a neurochemical evidence[J].Neurochem Res,2007,32(9):1559-1565.
[18] 孟辉,冯华,王宪荣.大鼠脑损伤后GABA及其受体GABA_AR的变化[J].第三军医大学学报,2001,(7):836-837.
[19] Reeves T M,Lyeth B G,Phillips L L,et al.The effects of traumatic brain injury on inhibition in the hippocampus and dentate gyrus[J].Brain Res,1997,757(1):119-132.
[20] Torcia M,Bracci-Laudiero L,Lucibello M,et al.Nerve growth factor is an autocrine survival factor for memory B lymphocytes[J].Cell,1996,85(3):345-356.
[21] Tian J,Chau C,Hales T G,et al.GABAA receptors mediate inhibition of T cell responses[J].Journal of Neuroimmunology,1999,96(1):21-28.
[22] Yoon S R,Ara C,Zixiong Z,et al.G-Aminobutyric acid promotes methionine-choline deficient diet-induced nonalcoholic steatohepatitis[J].J Biomed Res,2016,31(1):65-73.
[23] Lewis D A,Volk D W,Hashimoto T.Selective alterations in prefrontal cortical GABA neurotransmission in schizophrenia:a novel target for the treatment of working memory dysfunction[J].Psychopharmacology,2004,174(1):143-150.
[24] Castellano C,Cestari V,Cabib S,et al.Strain-dependent effects of post-training GABA receptor agonists and antagonists on memory storage in mice[J].Psychopharmacology,1993,111(2):134-138.
[2] Kennedy P G.Herpes simplex virus and the nervous system[J].Postgrad Medical Journal,1984,60(702):253-259.
[3] Nahmias A J,Lee F K,Beckman-Nahmias S.Seroepidemiological and sociological patterns of herpes simplex virus infection in the world[J].Scand J Infect Dis Suppl,1990,69(69):19-36.
[4] 周渊峰,周水珍,曹凌锋,等.儿童单纯疱疹病毒性脑炎 20 例临床分析及随访研究[J].中国实用儿科杂志,2011,26(7):527-530.
[5] Lellouch-Tubiana A,Fohlen M,Robain O,et al.Immunocytochemical characterization of long-term persistent immune activation in human brain after herpes simplex encephalitis[J].Neuropathol Appl Neurobiol,2000,26(3):285-294.
[6] Kapur N,Barker S,Burrows E H,et al.Herpes simplex encephalitis:long term magnetic resonance imaging and neuropsychological profile[J].J Neurol Neurosurg Psychiatry,1994,57(11):1334-1342.
[7] Costa E,Auta J,Grayson D R,et al.GABAA receptors and benzodiazepines:a role for dendritic resident subunit mRNAs[J].Neuropharmacology,2002,43(6):925-937.
[8] Bhat R,Axtell R,Mitra A,et al.Inhibitory role for GABA in autoimmune inflammation[J].Proc Natl Acad Sci U S A,2010,107(6):2580-2585.
[9] 衣曼,魏克伦,王雪峰,等.小鼠单纯疱疹病毒性脑炎模型的建立[J].小儿急救医学,2000,7(1):26-27.
[10] Satpathy G,Mishra A K,Tandon R,et al.Evaluation of tear samples for Herpes Simplex Virus 1 (HSV) detection in suspected cases of viral keratitis using PCR assay and conventional laboratory diagnostic tools[J].Br J Ophthalmol,2011,95(3):415-418.
[11] Ulett G C,Ketheesan N,Hirst R G.Cytokine gene expression in innately susceptible BALB/c mice and relatively resistant C57BL/6 mice during infection with virulent Burkholderia pseudomallei[J].Infect Immun,2000,68(4):2034-2042.
[12] Dirig D M,Yaksh T L.Intrathecal baclofen and muscimol,but not midazolam,are antinociceptive using the rat-formalin model[J].J Pharmacol Exp Ther,1995,275(1):219-227.
[13] Huse M,Lillemeier B F,Kuhns M S,et al.T cells use two directionally distinct pathways for cytokine secretion[J].Nat Immunol,2006,7(3):247-255.
[14] Hukkanen V,Broberg E,Salmi A,et al.Cytokines in experimental herpes simplex virus infection[J].Int Rev Immunol,2002,21(4-5):355-371.
[15] Katan M,Moon Y P,Paik M C,et al.Infectious burden and cognitive function:the Northern Manhattan Study[J].Neurology,2013,80(13):1209-1215.
[16] Gajcy K,Lochynski S,Librowski T.A role of GABA analogues in the treatment of neurological diseases[J].Curr Med Chem,2010,17(22):2338-2347.
[17] Goren M Z,Kucukibrahimoglu E,Berkman K,et al.Fluoxetine partly exerts its actions through GABA:a neurochemical evidence[J].Neurochem Res,2007,32(9):1559-1565.
[18] 孟辉,冯华,王宪荣.大鼠脑损伤后GABA及其受体GABA_AR的变化[J].第三军医大学学报,2001,(7):836-837.
[19] Reeves T M,Lyeth B G,Phillips L L,et al.The effects of traumatic brain injury on inhibition in the hippocampus and dentate gyrus[J].Brain Res,1997,757(1):119-132.
[20] Torcia M,Bracci-Laudiero L,Lucibello M,et al.Nerve growth factor is an autocrine survival factor for memory B lymphocytes[J].Cell,1996,85(3):345-356.
[21] Tian J,Chau C,Hales T G,et al.GABAA receptors mediate inhibition of T cell responses[J].Journal of Neuroimmunology,1999,96(1):21-28.
[22] Yoon S R,Ara C,Zixiong Z,et al.G-Aminobutyric acid promotes methionine-choline deficient diet-induced nonalcoholic steatohepatitis[J].J Biomed Res,2016,31(1):65-73.
[23] Lewis D A,Volk D W,Hashimoto T.Selective alterations in prefrontal cortical GABA neurotransmission in schizophrenia:a novel target for the treatment of working memory dysfunction[J].Psychopharmacology,2004,174(1):143-150.
[24] Castellano C,Cestari V,Cabib S,et al.Strain-dependent effects of post-training GABA receptor agonists and antagonists on memory storage in mice[J].Psychopharmacology,1993,111(2):134-138.