In this study, we investigated the effect of the extracts of Cyrtomium fortunei J.Sm. (CFJ) on lipopolysaccharide (LPS) induced inflammation in mouse BV-2 microglial cells. Nitric oxide (NO) production and cell viability were measured using the Griess reagent and the (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) (MTT) assay. Inflammatory cytokines were detected by quantitative polymerase chain reaction (qPCR) in BV-2 microglial cells with and without CFJ extracts. Subsequently, mitogen-activated protein kinases (MAPKs) and antioxidant markers were assessed by western blot analysis. It was found that the CFJ extract significantly decreased the production of pro-inflammatory cytokines (interleukin [IL]-6, tumor necrosis factor [TNF]-, and IL-1) and NO in BV-2 microglial cells that were stimulated with LPS. In addition, the expression levels of the phosphorylation of the MAPK family (p38, c-Jun N-terminal kinases [JNK], and extracellularsignal regulated kinase [ERK]) were reduced by CFJ. Also, treatment with CFJ significantly increased the activities of superoxide dismutase type 1(SOD1) and Catalase in BV-2 microglial cells. Our results indicate that CFJ has a potent suppressive effect on the pro-inflammatory responses of activated BV-2 microglia. Therefore, CFJ has the potential to be an effective treatment for neurodegenerative diseases, as it can inhibit the production of inflammatory mediators in activated BV-2 microglial cells.

본 연구는 동충하초 추출물을 PC12 및 BV2 세포에서 인지능력 개선에 대한 효능을 평가하고자 하였다. 동충하초 추출물을 증류수로 추출하여 PC12 및 BV2 세포로 MTT 분석을 통해 세포 생존율을 확인하고 L-glutamate로 유도한 PC12 세포를 통해 세포 보호 효능과 아세틸콜린 함량 및 아세틸콜린에스테아제 활성을 평가하였다. 또한, LPS로 유도한 BV2 세포를 통해 nitric oxide (NO) 및 prostaglandin E2 (PGE2) 생성량 등의 항염증 효능을 측정하고 western blot 분석을 통해 NK-ĸB, p38, JNK, caspase-3 등의 단백질 발현량을 확인하였다. 동충하초 추출물은 200 ㎍/㎖ 농도를 제외하고 1, 10, 100 (㎍/㎖) 농도에서 세포 독성이 나타나지 않았다. L-glutamate로 유도한 PC12 세포에서 유의성있게 세포 보호 효능과 아세틸콜린 함량의 증가, 아세틸콜린에스테아제 활성 감소가 나타났다. 또한, 동충하초 추출물은 NO 및 PGE2 생성량과 NK-ĸB, p38, JNK, caspase-3 등의 단백질 발현을 억제하였다. 이와 같은 결과는 동충하초 추출물이 인지능력에 대한 예방 및 개선 효능이 있음을 나타낸다. 따라서 동충하초 추출물은 인지능력 개선을 위한 새로운 천연 소재로 활용될 수 있다.

Taraxaci Herba has long been used in herbal medicine for their choleretic, anti-heumatic and diuretic properties. In the present study, we investigated the effects of origin plants of Taraxaci Herba, Taraxacum coreanum Nakai, as an anti-inflammatory agent in lipopolysaccharide(LPS)-induced microglial activation in BV2 cells. NNMBS273, the EtOH extracts of roots T. coreanum was examined for anti-neuronal inflammatory activity as new drug development. The roots of T. coreanum, showed the potent anti-neuroinflammatory effects on LPS-induced inflammation in microglial BV2 cells. The anti-inflammatory effects of NNMBS273, the EtOH extracts of roots T. coreanum was demonstrated by the suppression of pro-inflammatory mediators, including pro-inflammatory enzymes (inducible nitric oxide synthase and cyclooxygenase-2) and cytokines (tumor necrosis factor-α and interleukin-1β). These results suggest that the roots T. coreanum may be a promising candidate for the treatment of neurodegenerative diseases related to neuroinflammation.

[Background] Cordyceps militaris, a traditional medicinal mushroom, produces a component compound, cordycepin (3’-deoxyadenosine). Cordycepin has been known to have many pharmacological activities including immunological stimulating, anti-cancer, and anti-infection activities. However, the molecular mechanisms of inflammatory mediator’s activity by cordycepin remain poorly understood. In the present study, we investigat-ed the effects of cordycepin on the anti- inflammation cascades in lipopolysaccharide (LPS)-stimulated BV2 microglia cells. [Methods] Cordycepin were administered and their effects on LPS-induced pro-inflammatory mediators and MAP kinases were monitored by Western blotting and RT-PCR analysis. [Result] Cordycepin significantly inhibited the production of nitric oxide (NO), prostaglandin E2 (PGE2), and pro- inflammatory cytokines such as interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α in a concentration- dependent manner without causing cytotoxicity. Also, cordycepin suppressed inducible NO, synthase (iNOS) and cyclooxygenase-2 (COX-2) expression on the mRNA and protein level. In addition, cordycepin suppressed NF-κB translocation by blocking IkappaB- α (IκB-α) degradation and inhibited the phosphorylation of Akt, ERK-1/2, JNK, and p38 kinase. Our results indicate that the inhibitory effect of cordycepin on LPS -stimulated inflammatory mediator production in BV2 microglia is associated with the suppression of the NF-κB, Akt, and MAPK signaling pathways. Conclusion: Anti-inflammatory properties of cordycepin may be useful for treating the inflammatory and deleterious effects of microglial activation in response to LPS stimulation.