大鼠哺乳期环境烟草烟雾暴露对实验性自身免疫性脑脊髓炎发病的远期影响
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摘要
目的探讨大鼠哺乳期环境烟草烟雾(environmental tobacco smoking,ETS)暴露对实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)发病的远期影响。方法将36只SD乳鼠随机分成ETS-EAE(n=12)、ETS-假手术组(n=6)、空气-EAE组(n=12)和空气-假手术组(n=6)。于SD乳鼠出生后第2～21天,ETS-EAE组和ETS-假手术组给予ETS暴露,空气-EAE组(n=12)和空气-假手术组给予空气暴露。于SD乳鼠出生第11周时,采用豚鼠脊髓匀浆与完全福氏佐剂(CFA)混合乳液免疫诱导制备EAE模型,假手术组给予无菌生理盐水与CFA混合乳剂。从造模当天开始每天对大鼠进行神经系统行为学评分;于造模后第10、15、20天进行血浆γ干扰素(IFN-γ)水平测定;第20天时处死大鼠,通过HE染色和LFB染色定性分析脊髓中枢病理学改变。结果 ETS-EAE组大鼠神经系统行为学最高评分(3.58±0.37)、累积评分(27.25±3.41)及中枢病理HE评分(2.67±0.41)均高于其余组(均P<0.05);中枢神经系统炎性反应细胞数及脱髓鞘程度经定性分析较其他组升高;ETS-EAE组第10、15、20天血浆IFN-γ水平高于ETS-假手术组(均P<0.05);ETSEAE组第15天血浆IFN-γ水平高于空气-EAE组(P<0.05)。结论大鼠哺乳期ETS暴露可增加血浆IFN-γ水平,加重脊髓脱髓鞘程度及中枢炎性反应细胞浸润程度,加重成年后大鼠EAE发病。
Objective To investigate the distant effects of environmental tobacco smoking(ETS)exposure during lactation on experimental autoimmune encephalomyelitis(EAE).Methods 36 SD neonatal rats were randomly divided into ETS-EAE group(n=12),ETS-sham operation group(ETS-sham)(n=6),airEAE group(n=12)and air-sham operation group(air-sham)(n=6).ETS exposure was given in ETS-EAE group and ETS-sham group on the 2 nd to 21 st day after birth,while air-EAE group and air-sham group were exposed to pure air.In the 11 th week,the EAE model was induced by the mixture of guinea pig spinal cord homogenate and complete Freund's adjuvant(CFA),while the sham operation group was given sterile saline and CFA mixed emulsion.Rats were scored daily since the day of modeling,plasma interferon-gamma(IFN-γ)levels were measured on the 10~(th),15~(th) and 20~(th) days after modeling,and rats were sacrificed on the 20 th day.The pathological changes of spinal cord were observed by HE staining and LFB staining.Results The highest neurobehavioral score(3.58±0.37),cumulative score(27.25±3.41)and central pathological HE score(2.67±0.41)of the ETS-EAE group were higher than those of the other groups,all of the differences mentioned above were of statistical significance(all P <0.05);the number of inflammatory cells and demyelination degree of central nervous system of the ETS-EAE group were significantly higher than those of other groups by qualitative analysis;the plasma IFN-γlevel of the ETS-EAE group was higher than that of the ETS-sham operation group on the 10~(th),15~(th) and 20~(th) day(all P<0.05);the plasma level of IFN-γin the ETSEAE group was higher than that in the air-EAE group on the 15 th day,all the differences had statistical significance(P <0.05).Conclusions ETS exposure during lactation can increase plasma IFN-γlevel,aggravate the degree of demyelination of spinal cord and infiltration of central inflammatory cells,and aggravate the severity of EAE in adult rats.
Objective To investigate the distant effects of environmental tobacco smoking(ETS)exposure during lactation on experimental autoimmune encephalomyelitis(EAE).Methods 36 SD neonatal rats were randomly divided into ETS-EAE group(n=12),ETS-sham operation group(ETS-sham)(n=6),airEAE group(n=12)and air-sham operation group(air-sham)(n=6).ETS exposure was given in ETS-EAE group and ETS-sham group on the 2 nd to 21 st day after birth,while air-EAE group and air-sham group were exposed to pure air.In the 11 th week,the EAE model was induced by the mixture of guinea pig spinal cord homogenate and complete Freund's adjuvant(CFA),while the sham operation group was given sterile saline and CFA mixed emulsion.Rats were scored daily since the day of modeling,plasma interferon-gamma(IFN-γ)levels were measured on the 10~(th),15~(th) and 20~(th) days after modeling,and rats were sacrificed on the 20 th day.The pathological changes of spinal cord were observed by HE staining and LFB staining.Results The highest neurobehavioral score(3.58±0.37),cumulative score(27.25±3.41)and central pathological HE score(2.67±0.41)of the ETS-EAE group were higher than those of the other groups,all of the differences mentioned above were of statistical significance(all P <0.05);the number of inflammatory cells and demyelination degree of central nervous system of the ETS-EAE group were significantly higher than those of other groups by qualitative analysis;the plasma IFN-γlevel of the ETS-EAE group was higher than that of the ETS-sham operation group on the 10~(th),15~(th) and 20~(th) day(all P<0.05);the plasma level of IFN-γin the ETSEAE group was higher than that in the air-EAE group on the 15 th day,all the differences had statistical significance(P <0.05).Conclusions ETS exposure during lactation can increase plasma IFN-γlevel,aggravate the degree of demyelination of spinal cord and infiltration of central inflammatory cells,and aggravate the severity of EAE in adult rats.
引文
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[16]SpadaroM, GerdesLA, KrumbholzM, etal.Autoantibodies to MOG in a distinct subgroup of adult multiple sclerosis[J].Neurol Neuroimmunol Neuroinflamm,2016,3(5):e257.
[17]PrineasJW, RaineCS. Electronmicroscopyand immunoperoxidase studies of early multiple sclerosis lesions[J].Neurology,1976,26(6PT 2):29-32.
[18]Miljkovic D,Momcilovic M,Stojanovic I,et al.Astrocytes stimulate interleukin-17and interferon-gamma production in vitro[J].J Neurosci Res,2007,85(16):3598-3606.
[19]Nicholas JA,Boster AL,Imitola J,et al.Design of oral agents for the management of multiple sclerosis:benefit and risk assessment for dimethyl fumarate[J].Drug Des Devel Ther,2014,8:897-908.
[20]冷婷婷.干扰素γ及其受体与信号通路研究进展[J].甘肃科技纵横,2015,44(7):19-23.
[21] Gold R, Linington C, Lassmann H.Understanding pathogenesis and therapy of multiple sclerosis via animal models:70 years of merits and culprits in experimental autoimmune encephalomyelitis research[J].Brain,2006,129(Pt8):1953-1971.
[22]曹江晨,雷军平,李路军,等.辛伐他汀对变态反应性脑脊髓炎小鼠INF-γ和IL-10表达的影响[J].东南国防医药,2011,13(1):24-27.
[23]Oberg M,Jaakkola MS,Woodward A,et al.Worldwide burden of disease from exposure to second-hand smoke:a retrospective analysis of data from 192countries[J].Lancet,2011,377(9760):139-146.
[2]Gustavsen MW,Pang CM,Moen SM,et al.Environmental exposures and the risk of multiple sclerosis investigated in a Norwegian case-control study[J]. BMC Neurol,2014,14:196.
[3]Hedstr9m AK,Hillert J,Olsson T,et al.Smoking and multiple sclerosis susceptibility[J].Eur J Epidemiol,2013,28(11):867-874.
[4]杨亭亭,向雅娟,敖东慧,等.吸烟与多发性硬化和视神经脊髓炎谱系疾病发病风险关联性研究[J].中国现代神经疾病杂志,2016,16(9):598-602.
[5]Hedstr9m AK,Olsson T,Alfredsson L.Smoking is a major preventable risk factor for multiple sclerosis[J].Mult Scler,2016,22(8):1021-1026.
[6]Briggs FB,Gunzler DD,Ontaneda D,et al.Smokers with MS have greater decrements in quality of life and disability than non-smokers[J].Mult Scler,2017,23(13):1772-1781.
[7]Gallinat J,Meisenzahl E,Jacobsen LK,et al.Smoking and structural brain deficits:a volumetric MR investigation[J].Eur J Neurosci,2006,24(6):1744-1750.
[8]Procaccini C,Derosav V,Pucino V,et al.Animal models of multiple sclerosis[J].Eur J Pharmacol,2015,759:182-191.
[9]秦新月,余刚,董为伟,等.多发性硬化研究的动物模型制备[J].中国临床康复,2006,6(6):817-819.
[10]刘颖,刘华,辛晋敏.等.Wistar大鼠EAE模型的制备[J].长治医学报告,2005,19(1):58-60.
[11]Kono DH,Urban JL,Horvath SJ,et al.Two minor determinants of myelin basic protein induce experimental allergic encephalomyelitis in SJL/J mice[J].J Exp Med,1988,168(1):213-227.
[12]黄德晖,吴卫平,蒲传强,等.多发性硬化226例临床分析[J].中国神经免疫学和神经病学杂志,2003,10(3):152-155.
[13]Comi G,Radaelli M,Soelberg SP.Evolving concepts in the treatment of relapsing multiple sclerosis[J].Lancet,2017,389(10076):1347-1356.
[14]Wang ZW,Wang P,Lin FH,et al.Early-life exposure to lipopolysaccharide reduces the severity of experimental autoimmune encephalomyelitis in adulthood and correlated with increased urine corticosterone and apoptotic CD4+T cells[J].Neuroscience,2011,193:283-290.
[15]GenainCP, HauserSL. Experimentalallergic encephalomyelitis in the New World monkey Callithrix jacchus[J].Immunol Rev,2001,183:159-172.
[16]SpadaroM, GerdesLA, KrumbholzM, etal.Autoantibodies to MOG in a distinct subgroup of adult multiple sclerosis[J].Neurol Neuroimmunol Neuroinflamm,2016,3(5):e257.
[17]PrineasJW, RaineCS. Electronmicroscopyand immunoperoxidase studies of early multiple sclerosis lesions[J].Neurology,1976,26(6PT 2):29-32.
[18]Miljkovic D,Momcilovic M,Stojanovic I,et al.Astrocytes stimulate interleukin-17and interferon-gamma production in vitro[J].J Neurosci Res,2007,85(16):3598-3606.
[19]Nicholas JA,Boster AL,Imitola J,et al.Design of oral agents for the management of multiple sclerosis:benefit and risk assessment for dimethyl fumarate[J].Drug Des Devel Ther,2014,8:897-908.
[20]冷婷婷.干扰素γ及其受体与信号通路研究进展[J].甘肃科技纵横,2015,44(7):19-23.
[21] Gold R, Linington C, Lassmann H.Understanding pathogenesis and therapy of multiple sclerosis via animal models:70 years of merits and culprits in experimental autoimmune encephalomyelitis research[J].Brain,2006,129(Pt8):1953-1971.
[22]曹江晨,雷军平,李路军,等.辛伐他汀对变态反应性脑脊髓炎小鼠INF-γ和IL-10表达的影响[J].东南国防医药,2011,13(1):24-27.
[23]Oberg M,Jaakkola MS,Woodward A,et al.Worldwide burden of disease from exposure to second-hand smoke:a retrospective analysis of data from 192countries[J].Lancet,2011,377(9760):139-146.