氟西汀在实验性自身免疫性脑脊髓炎中的保护作用研究

Research for protective effect of Fluoxetine in experimental autoimmune encephalomyelitis

目的：探讨载脂蛋白2（Lipocalin2，LCN2）、CXCL10与实验性自身免疫性脑脊髓炎（Experimental autoimmune encephalomyelitis，EAE）的关系及氟西汀在EAE中的作用。方法：通过随机数字表将实验C57BL/6小鼠随机分为对照组、EAE组、干预组及氟西汀组，每组各20只。建模前经紫外线（280～320 nm）照射，干预组及氟西汀组建立EAE动物模型。于小鼠免疫第一天起氟西汀组小鼠连续给予氟西汀（10 mg/kg）灌胃；对照组、干预组和EAE组小鼠连续给予等量生理盐水灌胃，持续至实验结束。比较各组小鼠平均发病时间、神经功能评分、发病率；采用HE染色、免疫组织化学染色法观察各组小鼠脑组织病理变化；采用酶联免疫吸附法检测血清中CXCL10含量的变化并进行比较。结果：氟西汀组小鼠平均发病时间、神经功能评分、发病率均低于干预组、EAE组（ P＜0.05）。干预组低于EAE组（ P＜0.05）。氟西汀组小鼠脑组织中炎性反应较EAE组轻；LCN2免疫组化染色显示阳性细胞秩次明显低于EAE组（ P＜0.05）。血清中CXCL10含量氟西汀组较EAE组降低（ P＜0.05）。 EAE组LCN2水平与CXCL10含量相关。结论：LCN2、CXCL10与EAE存在重要联系，氟西汀可改善EAE临床症状，缓解EAE的发病情况。

To investigate the correlation between LCN 2 or CXCL10 and experimental autoimmune encephalomyelitis（EAE）,and the effect of Fluoxetine in EAE mice.Methods： Mice were randomly divided into four groups： control group, EAE group, intervention group and fluoxetine group , each group included twenty mice.EAE model were constructed with MOG35-55 in the intervention group and fluoxetine group ,following ultraviolet （UV） （280-320 nm） irradiation.Mice in fluoxetine group were given fluoxetine （10 mg/kg） by daily gavage since immunized ,and the saline was used in the control group ,intervention group and EAE group in the same way and same time.The drug/saline was continuously administered from the immunization to the day mice were sacrificed.The mean attacked time ,the nerve function grades and the incidence were observed and compared.We observed brain pathological changes by HE staining and immunohistochemistry;CXCL10 were tested by ELISA method ,and compared among the four groups.Results：The mean attacked time ,the nerve function grades and the incidence in the fluoxetine group were lower than those in the intervention group and EAE group ,there were significant differences between two groups （P〈0.05）,in the intervention group were lower than those in the EAE group ,there were significant differences between two groups （ P〈0.05 ）.Fluoxetine treated EAE mice showed decreased degree of inflammatory infiltration;the mean rank of positive cells lower in the fluoxetine group than those in the EAE group,the differences were significant between two groups （ P〈0.05 ）.The expression of CXCL10 in the peripheral blood plasma of fluoxetine group were lower than those in the EAE group ,there were significant differences between two groups （ P〈0.05 ）.Levels of LCN2 expression were correlated with those of CXCL 10 in the EAE group.Conclusion： LCN2, CXCL10 are correlated with the pathogenesis of EAE.Fluoxetine could alleviate clinical symptoms of EAE and alleviate the morbidity of EAE.