乌司他丁对小鼠脑脊髓炎神经再生作用研究

Effect of ulinastatin on the nerve regeneration of mice encephalomyelitis

目的探讨乌司他丁(ulinastatin,UTI)对小鼠实验性自身免疫性脑脊髓炎(experimental allergic encephalomyelitis,EAE)神经再生以及相关因子与蛋白表达的影响。方法 40只小鼠随机分为4组:乌司他丁组(U组)、阿托伐他汀对照组(A组)、空白对照组(C组)、正常对照组(N组),通过弗式完全佐剂和MOG35-55多肽构建小鼠EAE模型。分别于免疫3周、4周后进行HE、LFB和Bielschowsky染色,观察各组组织病理学变化。免疫组化法观察各组CD4+T细胞的表达数目。Western-blot检测4周后髓鞘碱性蛋白(myelin basic protein,MBP)、脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)、生长相关蛋白-43(growth associated protein-43,GAP-43)、2',3'-环核甘酸-3'-磷酸水解酶(2',3'-cyclic nucleotide-3'-phosphodiesterase,CNP)的表达情况。结果 U组神经功能最大分值低于C组,2者比较差异有统计学意义(P<0.05)。病理学特征显示U组炎症细胞、髓鞘脱失以及轴突损伤程度均轻于C组,且随着治疗时间的延长,神经损伤程度逐渐降低。UTI治疗后,MBP、BDNF、GAP-43、CNP表达均明显增高,与C组相比差异均具有统计学意义(P<0.05)。综合各方面指标发现UTI与阿托伐他汀治疗效果无明显差异。结论 UTI能有效减轻小鼠脑脊髓神经的残损程度,促进脑脊髓神经再生,为临床治疗人类多发性硬化(multiple sclerosis,MS)提供一定的理论基础。

Objective To investigate the effects of ulinastatin(UTI)on the nerve regeneration of mice experimental allergic encephalomyelitis and the expression of related factors and protein.Methods Forty mice were randomly divided into four groups:ulinastatin group(U),atorvastatin group(A), empty control group(C)and normal control group(N).The experimental autoimmune encephalomyelitis(EAE)in mice was constructed by Freund''s complete adjuvant and MOG35-55 polypeptide.Histopathological changes were observed by HE,LFB and Bielschowsky stained at the 3rd week and 4th week after immunized of each group.The expressions of CD4 +T cells were estimated by immunohistochemical method.The expression of myelin basic protein (MBP),brain-derived neurotrophic factor(BDNF),growth associated protein-43(GAP-43),2'',3''-cyclic nucleotide-3''-phosphodiesterase(CNP)were detected by Western-blot.Results The largest neurological score of group U was lower than group C,and the difference was statistically significant(P<0.05 ).Pathological features showed that the inflammatory cells,demyelination of spinal cord and axonal injury of group U were lighter than group C.With the duration of treatment,nerve injury decreased.After UTI treatment,the expression of MBP,BDNF,GAP-43,CNP increased.They were statistically significant difference when compared with group C(P<0.05).There was no significant difference between ulinastatin and atorvastatin in the treatment of EAE.Conclusion Ulinastatin could reduce the extent of nerve damage effectively and promote its regeneration which provide a theoretical basis for the clinical treatment of MS.