| Forsythiaside A has antalgic, antipyretic, antioxidative functions, antiviral, antibacterial pharmacological activities; is a effective active constituent of the Forsythia suspense, it regulate immune response by mediating various inflammation cytokines.Lipopolysaccharide is one of ingredients of the cell walls of gram-negative bacteria, can lead to inflammation response, are often used to copy inflammation model. When inflammation occurs, spleen take part in immune response as a peripheral immune organ, blood is an important carrier of various immune cells, where many immune response take place, so chicken and chicken spleen lymphocytes inflammation model by LPS in tthe research, under the action fo Forsythaside A, not only explore the change and relationship of inflammation cytokine of chicken spleen and spleen lymphocytes, but also relationship of antioxidant enzymes and free radicals from regulating inflammation cytocytes. Clarify the mechanisms on Forsythiaside A resist LPS in spleen of chicken, provide the theoretical basis for immune regulation of chicken.In vivo experiment,1day of eighty AA broilers were selected and randomly divided into four groups, respectively is:the normal control group, LPS group(200μg·kg-1), Forsythiaside A low dose treatment group[Forsythiaside A (30mg·kg-1)+LPS(200μg·kg-1)], Forsythiaside A high dose treatment group[Forsythiaside A (30mg·kg-1)+LPS(200μg·kg-1)],20in each group,when the15days, Forsythiaside A groups were given by gavage administration once a day for7days, control group and LPS group were given by gavage administration saline.until21days,200μg·kg-1BW-LPS were injected intravenously in Forsythiaside A groups and LPS group. two hours later,blood were adopted and preserve, chickens were killed, collecting the spleen and blood, observe clinical symptoms and pathological changes, calculate spleen organ index, on the other hand,some inflammation factor content and iNOS, NF-κB mRNA expression level by ELISA and qRT-PCR. At the same time, T-SOD, GSH, T-AOC, MDA, NO were tested.In vitro experiment,50days of A A broilers were selected, chickens were killed, collecting the spleen sterilely, by lymphotocyte separation medium, spleen lymphocytes were collected and counted and divided into20groups(4doses by five points).four groups include the control normal group, LPS injection group(5μg·mL-1), Forsythiaside A high dose treatment group[Forsythiaside A (400μg·kg-1)+LPS(5μg·mL-1)], Forsythiaside A low dose treatment group[Forsythiaside A (200 μg·kg-1)+LPS(5μg·mL-1)],20in each group,five points are:0h,6h,12h,24h,48h.method and indicator as in the above case.The results that clinical symptoms, spleen organ index and pathological changes shows that LPS can lead to increase body temperature, diarrhea, loss of appetite,drinking much water, spleen organ index incrases,while chicken in Forsythiaside A groups decrease spleen organ index and lymphocytes decrease in white pulp, HO increase,in the intervene of Forsythiaside A. it can reverse the above phenomenon.ELISA results shows that content of inflammation factor of spleen, blood and spleen lymphocytes increase in LPS groups compared with control group,IL-1β, IL-6, IL-8,1L-10, IL-17,TNF-α,PGE2,COX-2,iNOS,NO,TGF-β1,MIF,STAT-3,NF-Kb,INF-γ. while Forsythiaside A decrease content of above inflammation factor.decrease inflammation response.show positive correlation with inflammation factor change of spleen and spleen lymphocytes, through inhibiting inflammation factor of spleen and blood in the chicken by LPS, reduce inflammation reactions.Real time-PCR results shows that mRNA expression level of inflammation factor of spleen, blood and spleen lymphocytes increase in LPS groups compared with control group,IL-1β, IL-6, IL-8,IL-10,IL-17,TNF-a,PGE2,COX-2,iNOS,NO,TGF-p4,MIF,STAT-3,NF-κB,INF-y.while Forsy-thiaside A decrease mRNA expression level of above inflammation factor.decrease inflammation response.show positive correlation with inflammation factor change of spleen and spleen lymphocytes, through inhibiting inflammation factor of spleen and blood in the chicken by LPS, reduce inflammation reactions.It was showed that Antioxidant enzyme and free oxidical contents:Compared with the control normal group, the content of GSH, T-SOD and T-AOC of LPS group obviously decreased (P<0.05), showed that LPS can reduce Antioxidant enzyme content and activity to affect antioxidant function. Compared with the LPS group chicken, MDA content was significantly decreased (P<0.05), the content of GSH, T-SOD and T-AOC of the Forsythiaside A treatment group was significantly increased (P<0.05). The results shows that the Forsythiaside A can regulate antioxidative system, increase antioxidant capacity of spleen lymphocyte, spleen and blood.The results that ELISA and Real time-PCR show that Forsythiaside A perhaps decrease1L-1β,IL-8,IL-10,IL-17,IL-23,TNF-a,iNOS,INF-y content and mRNA expression level through NF-κB-COX-2-PGE2and IL-6/STAT-3signaling pathways, further affect migration of TGF-β4,TGF-β1and MIF, in the end play an antioxidant function. The results that antioxidant data show that Forsythiaside A perhaps improve T-AOC,T-SOD,GSH antioxidant enzymes and reduce MDA content, reach decrease oxidative damage.In summary, Forsythiaside A can obviously inhibit the expression and content of inflammation cytokines of spleen and blood and spleen lymphocytes induced by LPS, affect immune function of spleen and blood,shows that content of inflammatory cytokines of spleen and blood decrease, antioxidant enzyme content increase, free radical decrease, and change tendency is consistent, and it is positively correlated with spleen and blood antioxidant index and anti-inflammatory data, it’s function with dose and time, this shows Forsythiaside A and LPS were added to chicken, immune system was activated, it play an important biological effect, so as to adapt to immune mechanism, eventually have antioxidant and anti-inflammatory function. |
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