Constituentsoftheanti-asthmaherbalformulaASHMITMsynergisticallyinhibitIL-4andIL-5secretionbymurineTh2memorycells,andeotaxinbyhumanlungfibroblastsinvitro

<正>OBJECTIVE:Anti-asthma herbal medicine intervention（ASHMI ）,a combination of three traditional Chinese medicinal herbs developed in our laboratory,has demonstrated efficacy in both mouse models of allergic asthma,and a double-blind placebo-controlled clinical trial in patients with asthma.This study was designed to determine if the anti-inflammatory effects of individual herbal constituents of ASHMI? exhibited synergy. METHODS:Effects of ASHMI and its components aqueous extracts of Lingzhi（Ganoderma lucidum）,Kushen（Sophora flavescens） and Gancao（Glycyrrhiza uralensis）,on Th2 cytokine secretion by murine memory Th2 cells（D10.G4.1） and eotaxin-1 secretion by human lung fibroblast（HLF-1） cells were determined by measuring levels in culture supernatants by enzymelinked immunosorbent assay.Potential synergistic effects were determined by computing interaction indices from concentration-effect curve parameters. RESULTS:Individual Lingzhi,Kushen and Gancao extracts and ASHMI（the combination of individual extracts） inhibited production of interleukin（IL）-4 and IL-5 by murine memory Th2 cells and eotaxin-1 production by HLF-1 cells.The mean 25%-inhibitory-concentration(IC25) values（mg/mL） for ASHMI,Lingzhi,Kushen and Gancao for IL-4 production were 30.9,79.4, 123,and 64.6,respectively;for IL-5 production were 30.2,263,123.2 and 100,respectively; for eotaxin-1 were 13.2,16.2,30.2,and 25.1,respectively.The IC50 values（mg/mL） for ASHMI,Lingzhi,Kushen and Gancao for IL-4 production were 158.5,239.9,446.7,and 281.8, respectively;for eotaxin-1 were 38.1,33.1,100,and 158.5,respectively.The interaction indices of ASHMI constituents at IC25 were 0.35 for IL-4,0.21 for IL-5 and 0.59 for eotaxin-1.The interaction indices at IC50 values were 0.50 for IL-4 and 0.62 for eotaxin-1 inhibition.Inhibition of IL-5 did not reach IC50 values.All interaction indices were below 1 which indicated synergy. CONCLUSION:By comparing the interaction index values,we find that constituents in ASHMI? synergistically inhibited eotaxin-1 production as well as Th2 cytokine production.