神经药理学报 ›› 2018, Vol. 8 ›› Issue (2): 26-26.
• 2018年全国老年痴呆与认知障碍相关疾病学术会议论文摘要—基础研究 • 上一篇 下一篇
MENG Jie,NI Jun-jun,WU Zhou,ZHU Ai-qin,QING Hong,Hiroshi Nakanishi
摘要: Objective:Rheum tanguticum Maxim. ex Balf.( Rt),a traditional Tibetan medicine,is known to exert various bioactivities,including anti-inflammatory and anti-oxidative activities. The present study was conducted to investigate anti-infl ammatory and anti-oxidative effects of Rt on activated microglia. Methods:①MG6 microglial cell line,primary microglia and mice brain slices culture were stimulated by chromogranin A( CGA),an endogenous microglia activator,to induce proinfl ammation and oxidation. The anti-infl ammatory and anti-oxidative effects of Rt were examined by q-PCR,western blotting and ELISA by checking infl ammatory and oxidative molecules,including IL-1β,tumor necrosis factor-α,IL-10 and nitric oxide. ②Investigation of the involvement of IL-10 in the anti-infl ammatory effects of Rt on infl ammatory responses of microglia by using neutralizing antibodies against IL-10. Pro-infl ammatory molecules expression were checked by q-PCR.③Examination of the effects of major components of Rt( aloe-emodin,(+)-cathechin,piceatannol chrisophanol,physcion,β-sitosterol and emodin) on the IL-10 mRNA expression in MG6 cells. Results:①The mean mRNA expression levels of TNF-α,IL-1β and iNOS were signifi cantly increased in MG6 cells at 24 h after stimulation with CGA. Rt signifi cantly suppressed the mean mRNA expression levels of TNF-α,IL-1β and iNOS in GGA-stimulated MG6 cells. Furthermore,the production of IL-1β and NO metabolites in the culture medium of MG6 cells were assessed by using ELISA and NO2-/NO3- assay,respectively. The mean levels of IL-1β and NO2-/NO3- signifi cantly increased in the culture medium of MG6 cells at 24 h after treatment with CGA. Rt also signifi cantly decreased the mean levels of IL-1β and NO2-/NO3- in the culture medium of MG6 cells. The mean protein level of IL-1β signifi cantly increased in the organotypic hippocampal slice cultures at 48 h after stimulation with CGA( 10 nmol·L-1). Rt signifi cantly suppressed the mean protein level of IL-1β in CGA-stimulated organotypic hippocampal slice cultures ②The neutralization of IL-10 restored the mean mRNA expression of TNF-α and IL-1β in CGA-stimulated MG6 cells in the presence of Rt. ③Among the major components of Rt,three components namely, aloe-emodin (10 μmol·L-1),(+)-cathechin (30 μmol·L-1) and piceatannol (100 μmol·L-1),were found to signifi cantly increase the mRNA expression of IL-10 in MG6 cells at 24 h after treatment. In contrast, chrisophanol,physcion,β-sitosterol or emodin at concentrations up to 100 μmol·L-1 had no effect on the mRNA expression of IL-10 in MG6 cells. Conclusion:Rt suppressed the production of pro-infl ammatory and oxidative mediators,including IL-1β,TNF-α and NO,by cultured activated microglia through the production of IL-10. Two components of Rt,aloe-emodin and (+)-catechin,induced the secretion of IL- 10 from cultured microglia. Therefore,aloe-emodin and (+)-catechin are deemed responsible for the antineuroinfl ammatory and anti-oxidative effects of Rt through the secretion of IL-10 from microglia.