血清淀粉样P物质在EV71感染小鼠肺和脑表达及其意义初探
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摘要
目的探讨2周龄BALB/c小鼠在EV71感染后,不同分期肺和脑组织中血清淀粉样P物质(SAP)表达水平。方法 2周龄BALB/c小鼠按笼号分组,实验组35只,对照组29只。实验组予1×10~7TCID50·m L~(-1)的EV71病毒液0.1 mL腹腔注射,对照组腹腔注射等容量PBS液。观察小鼠的活动状态、皮疹、反应性、毛色、肢体活动及瘫痪情况等,依据实验组感染症状分为5期:精神反应期(建模后第2天)、部分瘫痪期(第3天)、全身瘫痪期(第3天)、死亡(第3~4天)和恢复期(第10天仍存活)。分别于5个分期称重实验组和对照组小鼠后处死,采集肺和脑组织以ELISA法测定SAP水平,分析不同分期SAP水平变化趋势。结果 1实验组建模后第2天均出现精神萎靡、活动减少,第3天部分小鼠出现肢体运动不协调、部分肢体瘫痪,甚至全身瘫痪至死亡,提示EV71感染小鼠模型建立成功。2建模后实验组精神反应期和对照组的体重差值差异无统计学意义,在部分瘫痪期、全身瘫痪期、死亡期和恢复期,两组体重差值的差异均有统计学意义。3实验组肺和脑组织中SAP水平在5个分期表达量均与对照组差异有统计学意义(P<0.05),实验组脑组织SAP水平是对照组的2~9倍,肺组织SAP水平是对照组的1.5~8倍。实验组部分瘫痪期、全身瘫痪期和死亡期脑组织SAP水平均显著高于精神反应期(P<0.05),肺组织SAP水平与精神反应期差异无统计学意义;脑和肺组织恢复期SAP水平均较精神反应期显著下降,但高于对照组水平。结论 SAP在EV71感染后小鼠的脑和肺组织中高表达,可能作为EV71感染后病情严重程度及预后判断的指标。
Objective To investigate the levels of serum amyloid P component in the lung and brain tissues in different stages in fourteen-day old BALB / c suckling mice infected with EV71. Methods Mice were randomly divided into two groups,the experimental group with 35 suckling mice and the control group with 29 suckling mice. Animals in the experimental group were injected intraperitoneally with 0. 1 m L 1 × 10~7TCID_(50)/ m L EV71 stocks,and those in the mock control group were mock-infected with an equal amount of phosphate-buffered saline( PBS). Mice were monitored daily to observe activity,rash,reactivity,hair color,physical activity and paralysis,and experimental group was divided into 5 stages according to the signs and symptoms of infection.Mental reaction period( day 2),partial paralysis period( day 3),paralysis stage( day 3),death( 3- 4 days) and the convalescent phase( on day 10 mice were still alive). 6- 8 mice that showed lethargy,weakness,limb shake,or paralysis were chosen,and killed upon symptom onset. Other survivors were monitored for up to 10 days and then killed. Tissue samples,including brain and lung,were collected from the mice. Levels of SAP were determined by enzyme-linked immunosorbent assay( ELISA) and the changing trend of SAP levels in different stages was analyzed. Results 1Mice showed less activity on day 2 postinfection and some were limb shake,some even paralysis to death on day 3 post-infection. It suggested that EV71 infected mice model was established successfully. There was no significant difference in body weight between the experimental group and the control group during the period of mental reaction,but the difference was statistically significant between two groups in the period of partial paralysis,paralysis,death and recovery period. 3 There was significant difference of SAP levels in both lung and brain tissues between the experimental groups and controls in five stages( P < 0. 05). In the experimental group,level of brain SAP was2- 9 times than that in the control group,and level of SAP in lung tissues was 1. 5- 8 times than in the control group. Brain SAP levels in experimental group in partial paralysis,paralysis period and period of death were significantly higher than those in mental reaction period( P < 0. 05). But it showed no statistical significance in lung between partial paralysis,paralysis,death and the convalescent. Levels SAP of recovery period decreased significantly in brain and lung tissue than those in period of mental reaction,but still higher than the level of the control group. Conclusion SAP is highly expressed in the brain and lung tissues of suckling mice infected with EV71,and it may be used as an index for judging the severity and prognosis of EV71 infection.
Objective To investigate the levels of serum amyloid P component in the lung and brain tissues in different stages in fourteen-day old BALB / c suckling mice infected with EV71. Methods Mice were randomly divided into two groups,the experimental group with 35 suckling mice and the control group with 29 suckling mice. Animals in the experimental group were injected intraperitoneally with 0. 1 m L 1 × 10~7TCID_(50)/ m L EV71 stocks,and those in the mock control group were mock-infected with an equal amount of phosphate-buffered saline( PBS). Mice were monitored daily to observe activity,rash,reactivity,hair color,physical activity and paralysis,and experimental group was divided into 5 stages according to the signs and symptoms of infection.Mental reaction period( day 2),partial paralysis period( day 3),paralysis stage( day 3),death( 3- 4 days) and the convalescent phase( on day 10 mice were still alive). 6- 8 mice that showed lethargy,weakness,limb shake,or paralysis were chosen,and killed upon symptom onset. Other survivors were monitored for up to 10 days and then killed. Tissue samples,including brain and lung,were collected from the mice. Levels of SAP were determined by enzyme-linked immunosorbent assay( ELISA) and the changing trend of SAP levels in different stages was analyzed. Results 1Mice showed less activity on day 2 postinfection and some were limb shake,some even paralysis to death on day 3 post-infection. It suggested that EV71 infected mice model was established successfully. There was no significant difference in body weight between the experimental group and the control group during the period of mental reaction,but the difference was statistically significant between two groups in the period of partial paralysis,paralysis,death and recovery period. 3 There was significant difference of SAP levels in both lung and brain tissues between the experimental groups and controls in five stages( P < 0. 05). In the experimental group,level of brain SAP was2- 9 times than that in the control group,and level of SAP in lung tissues was 1. 5- 8 times than in the control group. Brain SAP levels in experimental group in partial paralysis,paralysis period and period of death were significantly higher than those in mental reaction period( P < 0. 05). But it showed no statistical significance in lung between partial paralysis,paralysis,death and the convalescent. Levels SAP of recovery period decreased significantly in brain and lung tissue than those in period of mental reaction,but still higher than the level of the control group. Conclusion SAP is highly expressed in the brain and lung tissues of suckling mice infected with EV71,and it may be used as an index for judging the severity and prognosis of EV71 infection.
引文
[1]陈冉冉,何颜霞.肠道病毒71型感染致中枢神经系统损害预后的研究进展.中华实用儿科临床杂志,2015,30(4):318-320
[2]马慧敏,刘洪波,李运千,等.肠道病毒71型感染死亡患儿清道夫受体B族2型表达的意义.中华实用儿科临床杂志,2014,29(18):1393-1396
[3]Christian KA,Ijaz K,Dowell SF,et al.What we are watching--five top global infectious disease threats,2012:a perspective from CDC's Global Disease Detection Operations Center.Emerg Health Threats J,2013,6:20632
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[8]Wang SM,Lei H Y,Liu CC.Cytokine immunopathogenesis of enterovirus 71 brain stem encephalitis.Clin Dev Immunol,2012,2012:876241
[9]Deban L,Jaillon S,Garlanda C,et al.Pentraxins in innate immunity:lessons from PTX3.Cell Tissue Res,2011,343(1):237-249
[10]Deban L,Bottazzi B,Garlanda C,et al.Pentraxins:multifunctional proteins at the interface of innate immunity and inflammation.Biofactors,2009,35(2):138-145
[11]Manfredi AA,Rovere-Querini P,Bottazzi B,et al.Pentraxins,humoral innate immunity and tissue injury.Curr Opin Immunol,2008,20(5):538-544
[12]Pepys MB,Rademacher TW,Amatayakul-Chantler S,et al.Human serum amyloid P component is an invariant constituent of amyloid deposits and has a uniquely homogeneous glycostructure.Proc Natl Acad Sci U S A,1994,91(12):5602-5606
[13]Wang Y,Guo Y,Wang X,et al.Human serum amyloid P functions as a negative regulator of the innate and adaptive immune responses to DNA vaccines.J Immunol,2011,186(5):2860-2870
[14]Tian JQ,Wang Y,Lin N,et al.Active immunization with glucose-dependent insulinotropic polypeptide vaccine influences brain function and behaviour in rats.Scand J Immunol,2010,72(1):1-7
[15]Job ER,Bottazzi B,Gilbertson B,et al.Serum amyloid P is a sialylated glycoprotein inhibitor of influenza A viruses.PLo S One,2013,8(3):e59623
[16]吴丽芬,范倩,陈风华,等.血清淀粉样蛋白酶P(SAP)在3种常见结缔组织病患者中的表达及其意义.中国医学创新,2014(30):124-126
[17]Mulder SD,Hack CE,van der Flier WM,et al.Evaluation of intrathecal serum amyloid P(SAP)and C-reactive protein(CRP)synthesis in Alzheimer's disease with the use of index values.J Alzheimers Dis,2010,22(4):1073-1079
[18]Xi D,Luo T,Xiong H,et al.SAP:structure,function,and its roles in immune-related diseases.Int J Cardiol,2015,187:20-26
[2]马慧敏,刘洪波,李运千,等.肠道病毒71型感染死亡患儿清道夫受体B族2型表达的意义.中华实用儿科临床杂志,2014,29(18):1393-1396
[3]Christian KA,Ijaz K,Dowell SF,et al.What we are watching--five top global infectious disease threats,2012:a perspective from CDC's Global Disease Detection Operations Center.Emerg Health Threats J,2013,6:20632
[4]Deng L,Jia H L,Liu C W,et al.Analysis of differentially expressed proteins involved in hand,foot and mouth disease and normal sera.Clin Microbiol Infect,2012,18(6):188-196
[5]Xing W,Liao Q,Viboud C,et al.Hand,foot,and mouth disease in China,2008-12:an epidemiological study.Lancet Infect Dis,2014,14(4):308-318
[6]Solomon T,Lewthwaite P,Perera D,et al.Virology,epidemiology,pathogenesis,and control of enterovirus 71.Lancet Infect Dis,2010,10(11):778-790
[7]李鸣,于建云,李桢,等.肠道病毒71型感染9例死亡病例尸检分析.昆明医科大学学报,2013(10):145-147
[8]Wang SM,Lei H Y,Liu CC.Cytokine immunopathogenesis of enterovirus 71 brain stem encephalitis.Clin Dev Immunol,2012,2012:876241
[9]Deban L,Jaillon S,Garlanda C,et al.Pentraxins in innate immunity:lessons from PTX3.Cell Tissue Res,2011,343(1):237-249
[10]Deban L,Bottazzi B,Garlanda C,et al.Pentraxins:multifunctional proteins at the interface of innate immunity and inflammation.Biofactors,2009,35(2):138-145
[11]Manfredi AA,Rovere-Querini P,Bottazzi B,et al.Pentraxins,humoral innate immunity and tissue injury.Curr Opin Immunol,2008,20(5):538-544
[12]Pepys MB,Rademacher TW,Amatayakul-Chantler S,et al.Human serum amyloid P component is an invariant constituent of amyloid deposits and has a uniquely homogeneous glycostructure.Proc Natl Acad Sci U S A,1994,91(12):5602-5606
[13]Wang Y,Guo Y,Wang X,et al.Human serum amyloid P functions as a negative regulator of the innate and adaptive immune responses to DNA vaccines.J Immunol,2011,186(5):2860-2870
[14]Tian JQ,Wang Y,Lin N,et al.Active immunization with glucose-dependent insulinotropic polypeptide vaccine influences brain function and behaviour in rats.Scand J Immunol,2010,72(1):1-7
[15]Job ER,Bottazzi B,Gilbertson B,et al.Serum amyloid P is a sialylated glycoprotein inhibitor of influenza A viruses.PLo S One,2013,8(3):e59623
[16]吴丽芬,范倩,陈风华,等.血清淀粉样蛋白酶P(SAP)在3种常见结缔组织病患者中的表达及其意义.中国医学创新,2014(30):124-126
[17]Mulder SD,Hack CE,van der Flier WM,et al.Evaluation of intrathecal serum amyloid P(SAP)and C-reactive protein(CRP)synthesis in Alzheimer's disease with the use of index values.J Alzheimers Dis,2010,22(4):1073-1079
[18]Xi D,Luo T,Xiong H,et al.SAP:structure,function,and its roles in immune-related diseases.Int J Cardiol,2015,187:20-26