Xianjun Sun1, Xiaohong Deng1, Waijiao Cai1, Wenwei Li2, Ziyin Shen1, Tinghui Jiang3, Jianhua Huang4. 1. Institute of Integrated Chinese and Western medicine, Huashan Hospital, Fudan University, Shanghai 200040, China. 2. Department of Integrated Chinese and Western medicine, Zhongshan Hospital, Fudan University, Shanghai 200032, China. 3. Department of Interventional Radiology, Putuo Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200050, China. Electronic address: jiangtinghui@sina.com. 4. Institute of Integrated Chinese and Western medicine, Huashan Hospital, Fudan University, Shanghai 200040, China. Electronic address: hjhzyj@aliyun.com.
Abstract
AIMS: Icariin (ICA) is a flavonoid isolated from certain plant species in the genus Epimedium, especially Epimedium brevicornum. Previous studies indicated that ICA has certain regulatory effects on some inflammatory diseases, and that ICA regulates the activity of glucocorticoid receptor (GR) and NF-κB. But the causal link between GR and NF-κB and other downstream pathways in effects of ICA remained elusive, therefore here we have investigated whether ICA could promote GR function, in turn, to regulate NF-κB and/or other factors to achieve its anti-inflammatory effect. MAIN METHODS: Inflammatory cell models were induced by lipopolysaccharide (LPS) in RAW 264.7 and HeLa cell line. Observation of GRα nuclear translocation by confocal laser scanning microscopy. GRα and inflammatory cytokines expression was detected by RT-qPCR, Western Blotting and ELISA. Co-immunoprecipitation technique was used to detect the binding of GRα to downstream transcription factors. GRα activity was blocked by GRα antagonist RU486, and GR downstream transcription factors including NF-κB, c-Jun, and Stat3 were silenced by corresponding RNA interference. KEY FINDINGS: In both inflammatory cell models, ICA decreased LPS-induced production of inflammatory cytokines (IL-6 and TNF-α). While ICA up-regulated the amount of GRα and promoted its nucleus translocation. The increased GRα in the nucleus by ICA bound more NF-κB, c-Jun, and Stat3. Blockade GRα and silence of NF-κB, c-Jun, and Stat3 expression partially abolished the anti-inflammatory effects of ICA. SIGNIFICANCE: Promoted GR function and the consequent inhibition of pro-inflammatory transcription factors contribute a main mechanism by which ICA exerts its anti-inflammatory effect.
AIMS: Icariin (ICA) is a flavonoid isolated from certain plant species in the genus Epimedium, especially Epimedium brevicornum. Previous studies indicated that ICA has certain regulatory effects on some inflammatory diseases, and that ICA regulates the activity of glucocorticoid receptor (GR) and NF-κB. But the causal link between GR and NF-κB and other downstream pathways in effects of ICA remained elusive, therefore here we have investigated whether ICA could promote GR function, in turn, to regulate NF-κB and/or other factors to achieve its anti-inflammatory effect. MAIN METHODS: Inflammatory cell models were induced by lipopolysaccharide (LPS) in RAW 264.7 and HeLa cell line. Observation of GRα nuclear translocation by confocal laser scanning microscopy. GRα and inflammatory cytokines expression was detected by RT-qPCR, Western Blotting and ELISA. Co-immunoprecipitation technique was used to detect the binding of GRα to downstream transcription factors. GRα activity was blocked by GRα antagonist RU486, and GR downstream transcription factors including NF-κB, c-Jun, and Stat3 were silenced by corresponding RNA interference. KEY FINDINGS: In both inflammatory cell models, ICA decreased LPS-induced production of inflammatory cytokines (IL-6 and TNF-α). While ICA up-regulated the amount of GRα and promoted its nucleus translocation. The increased GRα in the nucleus by ICA bound more NF-κB, c-Jun, and Stat3. Blockade GRα and silence of NF-κB, c-Jun, and Stat3 expression partially abolished the anti-inflammatory effects of ICA. SIGNIFICANCE: Promoted GR function and the consequent inhibition of pro-inflammatory transcription factors contribute a main mechanism by which ICA exerts its anti-inflammatory effect.