Study On The Anti-inflammatory Effect And Mechanism Of The Plant Extracts Xpp-1 And Shikonin

Objective:To explore the anti-inflammatory effect and related mechanism of the plant extracts Xpp-1 and Shikonin,provide a study basis for the development of anti-inflammatory drugs.Methods:Mouse peritoneal macrophages were stimulated with LPS as an in vitro inflammatory cell model for studying the anti-inflammatory effect of Xpp-1;Mouse peritoneal macrophages were stimulated with CpG-ODN as an in vitro inflammatory cell model for studying the anti-inflammatory effect of Shikonin.We detected the toxicity on the mouse peritoneal macrophages of Xpp-1 and Shikonin by MTT assay;We also detected the effect of Xpp-1 and Shikonin pretreatment on pro-inflammatory cytokines mRNA expression in inflammatory model cells by q-PCR and Xpp-1 pretreatment on pro-inflammatory cytokines protein expression in inflammatory model cells by ELISA;Furthermore,the effect of Xpp-1 and Shikonin on the activation of NF-?B and MAPK signal pathways in inflammatory model cells was evaluated by western blot analysis.In vivo,we detected the effect of Xpp-1 on the production of IL-1?,IL-6,TNF-?in the peripheral blood of LPS-induced inflammatory mice by ELISA;We also observed the effect of Xpp-1 on the injury of the lung tissue in LPS-induced inflammatory mice by pathological section;Furthermore,the effect of Xpp-1 on the survival rate of LPS-induced inflammatory mice within 72 hours was observed.Results:No obvious toxicity was detected on mouse peritoneal macrophages in the concentration range of 6.25-25?pM of Xpp-1 and 1-2.5?M of Shikonin.In LPS-stimulated mouse peritoneal macrophages,Xpp-1(12.5?M)could inhibit mRNA and protein expression of IL-1?,IL-6,TNF-?(P<0.05),and inhibit the phosphorylation of IKK?/? and NF-?B.In CpG-ODN stimulated mouse peritoneal macrophages,Shikonin(2?M)could inhibit mRNA expression of TNF-?,IL-1?,IL-12p40(P<0.05),and inhibit the phosphorylation of IKK?/?,NF-?B and JNK.In vivo,in the LPS-induced inflammatory mice,Xpp-1(5mg/kg)could inhibit the production of pro-inflammatory cytokines such as IL-1?,TNF-? and IL-6 in the peripheral blood(P<0.05),reduce the inflammatory cell infiltration of lung tissue and improve the survival rate of the mice within 72 hours(P<0.05).Conclusions:The plant extracts Xpp-1 inhibited the expression of the IL-1?,TNF-a and IL-6 in LPS-stimulated mouse peritoneal macrophages;and the mechanism may be related to inhibition of activation of NF-?B signaling pathway;Xpp-1 reduced the concetration of IL-1?,TNF-? and IL-6 in the peripheral blood and the injury of lung tissue of LPS-induced inflammatory mice,so the plant extracts Xpp-1 has anti-inflammatory effect.Shikonin inhibited the expression of the TNF-a,IL-1? and IL-12 mRNA in CpG-ODN-stimulated mouse peritoneal macrophages,and the mechanism may be related to inhibition of activation of NF-?B and MAPK signal pathways,so Shikonin has anti-inflammatory effect.