西维来司钠对实验性自身免疫性脑脊髓炎模型大鼠的影响研究

Effect of sivelestat on the experimental autoimmune encephalomyelitis model rats

目的研究西维来司钠对实验性自身免疫性脑脊髓炎(EAE)模型大鼠的保护作用及机制。方法将40只大鼠随机分为正常组、模型组和低、中、高剂量实验组,每组8只。模型组和实验组注射豚鼠脊髓免疫抗原构建EAE模型,正常组注射等量生理盐水;建模成功后,低、中、高剂量实验组灌胃给予4,8,12mg·kg^(-1)西维来司钠,模型组和正常组灌胃等量生理盐水,每日1次,连续干预14 d。观察各组大鼠的神经功能,用苏木精-伊红(HE)染色法观察大鼠脊髓组织病理变化,以酶联免疫吸附(ELISA)法及反转录-聚合酶链式反应(RT-PCR)法检测各组大鼠JAKs/SATAs通路活化情况。结果干预后第12～14天实验组大鼠神经功能评分低于模型组(P<0.05)。正常组、模型组和低、中、高剂量实验组大鼠脑组织中STAT3蛋白表达量分别为(8.24±0.68),(15.49±1.21),(13.23±0.78),(11.26±0.56),(9.52±0.36)pg·mL^(-1),脊髓组织中JAK2 mRNA相对表达量分别为0.11±0.02,0.66±0.05,0.46±0.03,0.37±0.03,0.22±0.01;随着西维来司钠干预浓度的升高,低、中、高剂量实验组大鼠神经功能评分、脊髓组织病理学评分及脑组织中STAT3蛋白表达量和脊髓组织中JAK2 mRNA相对表达量逐渐降低(P<0.05)。结论西维来司钠可减轻EAE模型大鼠的神经损伤程度,可能与抑制JAKs/SATAs通路活化有关。

Objective To explore the protective effect of sivelestat on experimental autoimmune encephalomyelitis（ EAE） model rats and its mechanism. Methods Forty rats were randomly divided into normal group,model group and low,middle,high dose experimental groups,8 rats in each group. Model group and experimental groups were injected with spinal cord immune antigen of guinea pigs to construct the EAE model,and normal group was injected with equal amount of normal saline. After injection,low,middle,high dose experimental groups were given 4,8,12 mg·kg^-1 sivelestat,model group and normal group were given the same amount of normal saline. All rats were intervened 1 time per day,for 14 d continuously. The neurological function of rats in each group was observed and evaluated. The pathological changes of rats spinal cord tissue were observed by hematoxylin eosin staining（ HE）,enzyme linked immunosorbent assay（ ELISA） and reverse transcription-polymerase chain reaction（ RT-PCR） were used to detect the activationof JAKs/SATAs pathway in each group. Results On 12-14 d after intervention,the neurological function scores of experimental groups were lower than those of model group（ P〈0. 05）. The expressions of STAT3 protein in brain tissues of normal group,model group and low,middle,high dose experimental groups were（ 8. 24 ± 0. 68）,（ 15. 49 ± 1. 21）,（ 13. 23 ± 0. 78）,（ 11. 26 ± 0. 56） and（ 9. 52 ± 0. 36） pg·mL^-1,the relative expressions of JAK2 mRNA in the spinal cord were 0. 11 ± 0. 02,0. 66 ± 0. 05,0. 46 ± 0. 03,0. 37 ± 0. 03,0. 22 ± 0. 01. With the increase of the concentration of sivelestat,the neurological function score,the pathological score of spinal cord tissue,the expression of STAT3 protein in brain tissue and the relative expression of JAK2 mRNA in the spinal tissue were decreased gradually in low,middle,high dose experimental groups（ P 0. 05）. Conclusion Sivelestat can reduce the degree of nerve damage in EAE model rats,which may be related to the inhibition of activation of JAKs/SATAs pathway.