The Effects Of GX-50 On NF-ΚB Anti- Inflammation Pathway Of Macrophage

Inflammation is a defensive response to the traumatic injury,moderate activation of the inflammatory immune system is conductive to clear the pathogen and promote tissue healing.However,excessive inflammatory response may lead to more serious self-injury and increase the incidence of many diseases and mortality.Previous studies mostly look for anti-inflammatory drugs basing on the perspective of humoral factors while the found of cholinergic anti-inflammatory pathway(CAP) give a new strategy to the treatment of inflammation.Comparing with humoral anti-inflammatory pathway,the treatment of inflammation through CAP has the character of organ protection ,quick effect and widely extent.However, the commonly usedα7nAChR agonists,such as nicotine and carbachol,they can reduce the inflammation,but at the same time,they also has some toxixity that inhibit the blood pressure,respiration and heart rate.So their use is limited.Thus the development of low toxicityα7nAChR agonists is very important for the treatment of inflammation in the study of CAP. Object:GX-50, an ingredient from Zanthoxylum, has been suggested to a potentialα7nAChR agonists by structure-based drug design methods. Our previous studies showed thatα7nAChR was involved in the neuroprotective effect of GX-50 on beta-amyloid (Aβ)25-35-induced neuronal damage in primary cultured cortical neurons. And GX-50 could bind to alpha7 neuronal nicotinic acetylcholine receptor of cortical neurons. In this study, we investigated the effect of GX-50 for the LPS-induced enhance of inflammatory factor and NF-κB inflammatory pathways.Methods:Macrophage were treated with different concentrations of GX-50, and the viability was measured by MTT assay.For finding the effect of GX-50 to the anti-inflammatory pathway of Macrophage, ELISA and real time PCR were used to detect the contents of TNF-αand IL-1β, EMSA was used to detect the content of NF-κB,and Western blot experiment was used to measure the phosphorylation of IKK, IκB and the expression of TLR2.Results:Macrophage were treated by different concentrations of GX-50, MTT assay showed that the cell viability had no remarkable change.Afer pretreat macrophage with GX-50 for 1h and treat it with LPS for 4h,we could find that GX-50 inhibited LPS-induced enchance of TNF-αand IL-1βand the expression of their genes,at the same time,GX-50 inhibited the expression of NF-κB, the phosphorylation of IKK, IκB.Conclusion:Our research revealed that GX-50, could inhibit cytokine release induced by LPS and inhibit the activity of NF-κB pathway in macrophage. However, further researches should be done to make it clear that how GX-50 bind to theα7nAChR in the surface of macrophages.