Effects of baicalin on expression of inflammatory cytokines and inflammatory signaling pathways in porcine macrophages infected with Pasteurella multocida
In this study,we explored the effect of different concentrations of baicalin on the expression of inflammatory factors in porcine macrophages(HD11)infected with Pasteurella multocida(HN-13 strain)and their relationship with baicalin and inflammatory signaling pathway.Western blot was used to detect the expression levels of three key proteins(p-ERK/ERK、p-p38/p38,p-JNK/JNK)in the mitogen-activated protein kinase(MAPK)signaling pathway in the HN-13 infected HD11 cells(0,30min,60min,90min).Enzyme-related immunosorbent assay(ELISA)and real-time quantitative fluorescent PCR(RT-qPCR)were used to analyze the effects of different doses of baicalin:L group(20μmol/L),M group(40μmol/L),H group(60μmol/L)on the secretion and mRNA expression of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6)in porcine macrophages infected with P.multocida,control group(Control)and model group(HN-13)were set up simultaneously.The ERK pathway inhibitor SCH772984 was used to block this pathway,ELISA and PCR(RT-qPCR)were used to detect the secretion and mRNA expression of TNF-α,IL-1β and IL-6 of baicalin on HD11 cells infected with HN-13.Four groups were set up:control group(control group),model group(HN-13),inhibition group(HN-13 + ERK pathway inhibitor SCH772984),baicalin and inhibition group(HN-13+60μmol/L baicalin+ERK pathway inhibitor SCH772984).The results showed that the expression of p-ERK/ERK protein in HN-13 infected HD11 cells increased gradually with the extension of infection time(P<0.01),compared with the 0 min group(HN-13).There was no significant change in the expression of p-p38/p38 and p-JNK/JNK proteins(P>0.05).Different doses of baicalin significantly inhibited the secretion and mRNA expression levels of TNF-α,IL-1β,IL-6(P<0.05),compared with model group(HN-13),and the inhibitory effect in a concentration-dependent manner.Compared with inhibition group,baicalin and inhibition group significantly inhibited the secretion and mRNA expression levels of TNF-α,IL-1β,IL-6(P<0.05).In conclusion,baicalin can inhibit the expression and secretion of ERK pathway related proteins in porcine macrophages(HD11)infected with P.multocida(HN-13 strain),thereby exerting anti-inflammatory effects.This study provides ideas for the research of new drugs for the prevention and treatment of swine pulmonary disease caused by P.multocida.
BaicalinPasteurella multocidaporcine macrophageinflammatory expressionMAPK signal path way
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