A Study Of A Methanolic Extract From Ophiocordyceps Sinensis Mycelia On Inhibition Of LPS-induced Inflammatory Response In Rat Alveolar Macrophages

Aim: Cordyceps sinensis was used in pulmonary diseases. Ophiocordycepssinensis(XhQ anamorph of Cordyceps sinensis),\r\recent years were found to bebenificial for some lung diseases,especially in pulmonary fibrosis prevention andinhibition. Preliminary study found Ophiocordyceps si’nensis benifits may beassociated with anti-inflammatory activity.We used LPS(l[xg/mL)to establish an inlfammatory response model on Ratalveolar macrophages NR8383cell line Then we studied the inhibitory mechamismsof methanolic extract from Ophiocordyccps sinensis mycelia (OSME)oninflammatory response. We used Western blot to detect the expression ofinflammatory proteins such as iNOS and COX-2. IkB phosphorylation and thedistribution change of NF-KB(critical signal molecular in NF-kB pathway) werealso detected.Result: In physiological condition,NR8383cells had a low expression of iNOSprotein.When stimulated with LPS(l|xg/mL)for3hours, a sharp increase of iNOSwas found (2.33times as the control group).Whereas,OSME(in concentration scale50-200ng/mL)could attenuate the high expression of iNOS. The ratio of relativeexpression decreased from2.33to0.91±0.088、0.56土0,067、0,54±0<.018(p0.05).The inhibition percentage at3h were59.22%、74.87%、75.67%.In physiological condition,there was a low expression of COX-2protein inNR8383cells.When stimulated with L.PS(l|.ig/mL) for3h a sharp increase ofCOX-2was found (10.17times as the control group).When cells were treated withOSME(in concentration scale50-200|.ig/mL).the high expression of COX-2wasattenuated (p<0.05). The ratio of relative expression decreased!rom10.17to7.26±0.971、5.90±3.21、5.3±3.87(p<0.05). The inhibition percentage at3h were28.61%、41.98%、47.86%. In physiological condition, transcription factor NF-kB mainly existed incytoplasm,inactive.When cells were treated with LPS(lfxg/mL) for15min，NF-kBtransferred to nucleus.a decrease of cytoplasmic NF-kB(0.52±0.365) and an increaseof nuclear NF-kB(1.64士0.278) were detected.Whereas,when cells were incubatedwith OSME (concentration50and100}xg/mL)，the translocation was slightlysuppressed. Cytoplasmic NF-kB rose to0.809±0.181、0.90土0.277，nuclear NF-kBdecreased to1.24土0.152、1.09±0.181(p<0.05).Study on IkB phosphorylation showed that LPS could increase relative p-IkBamount in cells from1to1.64土0.223，when treated withOSME(50-200jxg/mL),cellular p-IicB decreased to0.21士0.023、0.79士0.022、0.58土0.058倍（p<0.05).Conclusion:OSME could inhibit LPS induced cell inflammatory response bysuppressing the activity of NF-kB pathway.