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The Critical Role of IL-10 in the Anti-neuroinflammatory and Anti-oxidative Effects of Rheum Tanguticum on Activated Microglia
MENG Jie，NI Jun-jun，WU Zhou，ZHU Ai-qin，QING Hong，Hiroshi Nakanishi
2018, 8 (2):
26-26.
摘要
(
223 )
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.
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