Anti-inflammatory effects of extracts from Sanhuang-Siwu-Tang in different solvents
Objective To explore the in vitro anti-inflammatory effects and related mechanisms of different solvent extracts of Sanhuang-Siwu-Tang(SST),analyze the relationship between its main components and anti-inflammatory effects,and investigate its in vivo anti-inflammatory effects.Methods SST water extract(SSTW),as well as 95％ethanol extract(SSTE)and 50％ethanol extract(SSTWE)were prepared.High-Performance Liquid Chromatography(HPLC)was used to analyze the main components.An inflammatory model was established by inducing RAW264.7 cells with LPS,and a rat model of chronic pelvic inflammatory disease(CPID)was established by phenol mucilage method.Cell viability was determined by MTT;NO level was measured by Griess;PGE2,IL-6,and TNF-α levels were examined by ELISA;COX-2,iNOS,MAPKs,and NF-κB protein expressions were measured by Western blot;pathological changes in the rat uterus were observed through pathological sections.Results There was no significant difference in cell viability between the SSTWE groups at various concentrations from 0.78 to 25 μg/mL and normal group(P＞0.05).The anti-inflammatory effects of SSTWE at 12.5 p,g/mL were weaker than those of SSTE but similar to those of SSTW,while the anti-inflam-matory effects of SSTWE at 25 μg/mL were stronger than those of SSTW but similar to those of SSTE.SSTW,SSTE,and SSTWE all significantly reduced iNOS and COX-2 protein expressions in LPS-induced RAW264.7 cells(P＜0.05).SSTWE significantly reduced the protein expressions of p-ERK,p-IκB-α,and nuclear p65 in LPS-induced RAW264.7 cells,while increasing IκB-αtotal protein expression and cytosolic p65 protein expression(P＜0.05).SST significantly lowered serum TNF-α and IL-6 levels in CPID rats(P＜0.05),reduced local pelvic tissue adhesion,and improved pathological morphology of the uterus.Conclusion SST has significant in vitro anti-inflammatory effects,and its mechanisms are related to the inflammatory pathways of MAPKs and NF-κB.Its different extracts have varying anti-inflammatory effects and cytotoxicity,possibly related to the components of Danggui(Radix Angelicae Sinensis).Moreover,SST has significant anti-inflammatory effects on CPID in rats and can be considered as a candidate drug for CPID.
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