CAJ | 학술논문

目的：探讨丙戊酸钠（ VPA）对实验性自身免疫性脑脊髓炎（ EAE）大鼠发病的影响及其免疫调节机制。方法50只雌性SD大鼠随机分为5组：空白对照组、EAE组、VPA低剂量组（100 mg/kg）、VPA中剂量组（300 mg/kg）和VPA高剂量组（600 mg/kg）。采用豚鼠脊髓匀浆免疫大鼠建立EAE模型，从免疫当天开始每日2次分别给予腹腔注射上述3种不同剂量的VPA或生理盐水，直到发病高峰期处死。观察大鼠行为学变化进行临床症状评分；采用苏木素-伊红染色观察中枢神经系统（CNS）内炎性细胞浸润；采用免疫组化观察CNS内小胶质细胞表达；用酶联免疫吸附法（ELISA）检测CNS内IFN-γ、IL-17和IL-10的含量。结果与EAE组比较，大、中、小剂量VPA组大鼠EAE发病率下降，临床症状减轻，潜伏期延长，CNS内炎性细胞浸润减少，但仅中剂量VPA组在EAE临床症状改变方面差异有统计学意义（P＜0．05），而中剂量和高剂量组病理学改变差异均有统计学意义（P＜0．05）；免疫组化结果显示，与EAE组比较，中剂量和高剂量VPA组的活化的小胶质细胞数量显著减少（P＜0．05）；ELISA结果显示，与EAE组比较，VPA干预后，大脑和腰髓中的IL-10水平上升而IFN-γ和IL-17水平明显减少，中剂量VPA组最为显著（ P＜0．05）。结论 VPA对EAE大鼠具有神经保护作用，中剂量最为显著，其作用机制可能与免疫调节作用有关。
목적：탐토병무산납（ VPA）대실험성자신면역성뇌척수염（ EAE）대서발병적영향급기면역조절궤제。방법50지자성SD대서수궤분위5조：공백대조조、EAE조、VPA저제량조（100 mg/kg）、VPA중제량조（300 mg/kg）화VPA고제량조（600 mg/kg）。채용돈서척수균장면역대서건립EAE모형，종면역당천개시매일2차분별급여복강주사상술3충불동제량적VPA혹생리염수，직도발병고봉기처사。관찰대서행위학변화진행림상증상평분；채용소목소-이홍염색관찰중추신경계통（CNS）내염성세포침윤；채용면역조화관찰CNS내소효질세포표체；용매련면역흡부법（ELISA）검측CNS내IFN-γ、IL-17화IL-10적함량。결과여EAE조비교，대、중、소제량VPA조대서EAE발병솔하강，림상증상감경，잠복기연장，CNS내염성세포침윤감소，단부중제량VPA조재EAE림상증상개변방면차이유통계학의의（P＜0．05），이중제량화고제량조병이학개변차이균유통계학의의（P＜0．05）；면역조화결과현시，여EAE조비교，중제량화고제량VPA조적활화적소효질세포수량현저감소（P＜0．05）；ELISA결과현시，여EAE조비교，VPA간예후，대뇌화요수중적IL-10수평상승이IFN-γ화IL-17수평명현감소，중제량VPA조최위현저（ P＜0．05）。결론 VPA대EAE대서구유신경보호작용，중제량최위현저，기작용궤제가능여면역조절작용유관。
Objective To investigate the effects of valproic acid ( VPA ) on SD rats with experimental autoimmune encephalomyelitis ( EAE) and its possible immunomodulatory mechanism .Meth-ods Fifty female Sprague-Dawley ( SD) rats were randomly divided into five groups by random digit table , including control group (n=10), EAE group(n=10), low dose VPA treated group (100 mg/kg, n=10), median dose VPA treated group (300 mg/kg, n=10) and high dose VPA treated group (600 mg/kg, n=10).The SD rat model of EAE was induced by immunizing with a guinea pigs′spinal cord homogenate (GPSCH).Normal saline and various doses of VPA were given to rats in according groups twice a day from day 0 to day 19 ( close to the peak stage of EAE ) .The severity of EAE was scored according to the signs and symptoms.Pathological changes were observed through Hematoxylin-Eosin staining, and then the degrees of inflammatory infiltration were evaluated .The numbers of activated neuroglia that expressed Iba-1 in cerebral and lumber cords were counted by immunohistochemistry .The expression of IFN-γ, IL-17 and IL-10 in cer-ebral and lumber cords were measured by ELISA .Results Compared with EAE group , rats in the low, me-dian and high dose VPA treated groups had lower incidence of EAE and prolonged latency , but only the me-dian dose treated group showed significant alleviation in clinical symptoms (P<0.05).Both the median and the high dose treated group showed decreased inflammatory cell infiltration in CNS (P<0.05).Immunohisto-chemistry results showed that the numbers of activated microglia were significantly inhibited in rats treated with median and high dose of VPA in comparison with those in EAE group (P<0.05).Results of ELISA demonstrated that the expression of IFN-γand IL-17 in both median and high dose VPA treated groups were significantly decreased compared with those in EAE group (P<0.05), but only the median dose treated group showed a remarkably increased expression of IL-10 (P<0.05).Conclusion VPA, especially medi-um dose of VPA ( 300 mg/kg ) , had neuroprotective effects on rats with EAE .The possible mechanism might be associated with the inhibited activation of microglia and the increased percentage of anti -inflammato-ry cytokines .