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目的:观察实验性自身免疫性脑脊髓炎(EAE)小鼠模型脑组织和脊髓中脑型肌酸激酶(CK-BB)、钙泵(CaATPase)和钙中性蛋白酶(calpain)的变化。方法:C57BL/6小鼠随机分为:EAE组(n=9),用髓鞘少突胶质细胞糖蛋白35-55多肽(MOG35-55)及完全弗氏佐剂混合抗原乳剂免疫小鼠;对照组(n=5),注射生理盐水。用MOG35-55诱导C57BL/6小鼠,建立(EAE)动物模型(即多发性硬化模型)。观察记录EAE小鼠行为学变化;采用苏木精-伊红染色、LFB髓鞘染色,酶标仪和分光光度计法检测发病高峰期(免疫后第19天)的中枢神经组织病理学变化、CK-BB,CaATPase和calpain酶活性的改变。结果:EAE组与对照组相比:①日均神经功能评分和日累积评分增加(P<0.01);②苏木精-伊红染色:中枢炎症细胞浸润明显(P<0.05);③LFB染色:髓鞘脱失较多;④CK-BB和CaATPase活性降低,calpain活性增加(P<0.05);⑤EAE组小鼠(只)日均累积神经功能评分与CK-BB和CaATPase活性呈负相关(P<0.05);⑥EAE组小鼠(只)日均累积神经功能评分与calpain酶活性呈正相关(P<0.05)。结论:EAE高峰期中枢CK-BB和CaATPase活性降低、calpain活性增高是EAE发生发展的后果,提示EAE时中枢能量代谢和CaATPase、calpain的病理性损害。
OBJECTIVE: To observe the change of brain-type creatine kinase (CK-BB), calcium pump (CaATPase) and calpain in brain and spinal cord of experimental autoimmune encephalomyelitis (EAE) Methods: C57BL / 6 mice were randomly divided into EAE group (n = 9), mice immunized with mixed myelin oligodendrocyte glycoprotein 35-55 peptide (MOG35-55) and complete Freund’s adjuvant ; Control group (n = 5), saline injection. C57BL / 6 mice were induced with MOG35-55 to establish an animal model of (EAE) (ie multiple sclerosis model). The behavioral changes of EAE mice were observed and recorded. The histopathological changes of central nervous system were detected by hematoxylin-eosin staining, LFB myelin staining, microplate reader and spectrophotometer at the peak of onset (day 19 after immunization) CK-BB, CaATPase and calpain enzyme activity changes. Results: Compared with the control group, the daily mean score of neurological function and daily cumulative score were increased (P <0.01); ② hematoxylin-eosin staining: the infiltration of central inflammatory cells was significantly (P <0.05); ③LFB staining: (3) There was a significant negative correlation between the daily cumulative neurological score and the activity of CK-BB and CaATPase in EAE group (P <0.05) 0.05). (6) The daily average cumulative neurological score of EAE group mice was positively correlated with calpain enzyme activity (P <0.05). CONCLUSION: The decrease of central CK-BB and CaATPase activity in the peak of EAE and the increase of calpain activity are the consequence of the development of EAE, suggesting the central energy metabolism and the pathological damage of Ca ATPase and calpain in EAE.