Effect of ethanol extract of root of Anacyclus pyrethrum on regulating neuroinflammation in brain of cough variant asthma rats through JNK and p-JNK/p38MAPK signaling pathways
Objective To investigate the regulatory effects of(ethanol extract of the root of Anacyclus pyrethrum,EEAP)on neuroinflammation and c-Jun N-terminal kinase(JNK)/p38 mitogen-activated protein kinase(p38MAPK)signaling pathway in cough-variant asthma(CVA)rats.Methods In in vitro experiments,rat hippocampal neuronal cells H19-7 were divided into six groups,namely,the control group,the model group,the prednisone acetate(0.025 mg·mL-1)positive drug control group,and the EEAP high,medium,and low mass concentration(6.4,3.2,and 1.6 mg·mL-1)group,and lipopolysaccharide(LPS)was used to induced cellular inflammation model,and the levels of interleukin-6(IL-6),IL-8,and IL-18 in the cell supernatants of each group were detected by enzyme-linked immunosorbent assay(ELISA),and the protein expression of JNK,p-JNK,and p38MAPK in the neuronal cells of each group was detected by Western blotting.In the animal experiments,SPF-grade male SD rats were randomly divided into control,model,prednisone acetate(2.5 mg·kg-1),and EEAP high,medium,and low dose(640,320,and 160 mg·kg-1)groups,and rats in each group except the control group were sensitized with 1 mg of OVA,and the rats sc freshly prepared 1 mg·mL-1 OVA solution was injected at a total of 10 points in the hindfoot plantar,inguinal,lumbar,dorsal,and cervical regions of each rat,with 0.05 mL injected at each point,while 0.5 mL was injected intraperitoneally,for a total of 1 mL.Starting on the 15th d after modeling,the rats were continuously nebulized and inhaled with 1％OVA for 15 d,once per day,for 20 min each time in order to stimulate asthma.The control group was sc and nebulized inhalation with 0.9％NaCl solution instead of OVA solution.Starting from the 30th d of modeling,the control group and the model group ig were given equal amounts of 0.9％NaCl solution,and each dosing group ig was given the corresponding dose of drug once a day for 30 d.The expression of JNK and p38MAPK in hippocampal tissues was detected by immunohistochemistry,and the protein expression of p-JNK in the tissues of hippocampal region was detected by Western blotting.ELISA was used to detect the levels of IL-6,IL-8 and IL-18 in the hippocampal area tissues of rats in each group.Results The in vitro experiments showed that the levels of IL-6,IL-8,and IL-18 in cell culture supernatants of the model group were significantly higher compared with those of the control group(P＜0.05),and the protein expression levels of p-JNK and p38MAPK in LPS-stimulated neuronal cells in the model group were significantly higher(P＜0.05).Compared with the model group,the levels of IL-6,IL-8,and IL-18 in cell culture supernatants of the prednisone acetate group and the EEAP high-and medium-dose groups were significantly decreased(P＜0.05).The levels of IL-6,IL-8 and IL-18 in cell culture supernatant were significantly reduced(P＜0.05),and the protein expression levels of p-JNK and p38MAPK in cells were significantly reduced(P＜0.05).And the inhibitory effects of inflammatory factors and protein expression of EEAP showed concentration correlation.In animal experiments,the levels of IL-6,IL-8,IL-18 and the protein expression levels of JNK,p-JNK and p38MAPK in the brain tissues of rats in the model group were elevated compared with those in the control group(P＜0.05).Compared with the model group,the levels of IL-6,IL-8,IL-18 and the protein expression levels of JNK,p-JNK and p38MAPK were decreased in the prednisone acetate group and the EEAP groups(P＜0.05).And the EEAP effect was dose-related.Conclusion EEAP inhibited CVA-induced neuroinflammation and could exert anti-inflammatory effects by regulating inflammatory factors IL-6,IL-8,IL-18 and JNK,p-JNK/p38MAPK signaling pathways.
ethanol extract of Anacyclus pyrethrumcough variant asthmaneuroinflammatory responsec-Jun N-terminal kinase/p38 mitogen-activated protein kinase signaling pathwayinterleukinshippocampal neurons
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