The Study Of Anti-inflammatory Activities And Mechanism Of Aloe-emodin In Vitro

Objective:We set up the inflammatory mode with RAW264.7macrophage line induced by LPS. Aloe-emodin was studied to examine the release of NO, TNF-a,IL-1β and to determin the protein levels of ERK,iNOS in RAW264.7induced by LPS. It will provide some experiment foundations for further studies.Methods:MTT assay was adopted to prepare the growth curve of RAW264.7.Nitric oxide(NO) production was examined by Griess reaction; TNF-a and IL-1β were detected by enzyme linked immunosorbent assay (ELISA). The protein levels of ERK,iNOS was studied by Western blot.Transient transfection was used to study the expression of NF-kappa B activation in reporter gene.Results:1. After treatment with Aloe-emodin0.1μM-40μM on RAW264.7for24h,compared with vehicle group,no obvious effect was fund(P>0.05).2.BAY-11-708210μM,Aloe-emodin20μM,emodin40μM and Rhein20μM can decrease the release of NO in RAW264.7induced by LPS, compared with the model group, The difference was statistically significant (P<0.01).On the other hand,the inhibition of the release of NO by8μM Physcion and8uM Chrysophanol is not obvious, and there were no significant differences with the model group.3. Compared with model group, Aloe-emodin20μM,10μM as well as BAY11-7082were able to significantly inhibit the release of NO,TNF-α, and IL-1β,compared with the model group, The difference was statistically significant (P<0.01),however the inhibition of Aloe-emodin5μM was not significant (P>0.05).4.1μg/ml LPS could stimulate the expression of iNOS and ERK protein in RAW264.7. The experimental results showed that Aloe-emodin can inhibit the expression of iNOS and ERK protein induced by LPS in RAW264.7cells.5. The expression of NF-kappa B was stimulated by TNF-a in HELA cell, there was significant difference between model group and blank group (P<0.01), BAY11-7082can significantly inhibit the expression of NF-kappa B in HELA cells stimulated by TNF-a, compared with model group,the difference was statistically significant (P <0.01). But different concentrations of Aloe-emodin could not.Conclusion:In summary, this experimental study found that Aloe-emodin could reduce the release of NO TNF-a, and IL-1β,and inhibit the expression of iNOS in RAW264.7cells induced by LPS in dose-dependent manner, which may have a relationship with the ERK path way.