Effect of pretreatment with puerarin on activation of LPS-induced RAW264.7 cells

2012 
Objective: To observe the effect of pretreatment with puerarin on activation of LPS-induced RAW264.7 cells and secretory cytokines,and discuss its anti-inflammatory mechanism.Method: Well-grown RAW264.7 cells in the exponential phase were collected and randomly divided them into the blank control group,the LPS group and the puerarin pretreatment+LPS group.The cellular toxic effect of puerarin on RAW264.7 cells was examined by CCK-8 assay,cell morphology was detected by Giemsa stain method,the changes in TNF-α and MIP-2 were tested by ELISA,and the expression of NF-κB p65 mRNA were determined by qRT-PCR.Result: When puerarin was cultured with 1 mg·L-1 LPS at a concentration of lower than 400 μmol·L-1,it had not showed the cellular toxic effect(P0.05).Compared with the control group,the LPS group could significantly change the morphology of RAW264.7 cells(increase in cell body,irregular shape,with a large number of pseudopodia extending).After intervention,the puerarin 100 μmol·L-1 group could significantly inhibit LPS-induced cell morphological changes,while the puerarin 200 μmol·L-1 and 400 μmol·L-1 puerarin groups showed more notable inhibitory effects.However,there was no obvious difference between the two groups.The pretreatment with puerarin could inhibit the expression of TNF-α and MIP-2 in cell supernatant and NF-κB p65 mRNA in cells(P0.05).With increase in the puerarin concentration,its inhibitory effect gradually grew(P0.05),but did not reach the level of the blank control group.Conclusion: As a safe and effective natural anti-inflammatory drug,puerarin can significantly reduce the expression of inflammatory cytokines(TNF-α,MIP-2).Its mechanism may be related to the reduction of NF-κB p65 mRNA expression.
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