Intervention Effect And Mechanism Of PCPC On AFB1-induced Immunosuppression In Broilers Based On JAK-STAT Signaling Pathway

With the development of the poultry industry,the breeding density of poultry has gradually increased,but the chicks are yin and yang,which are easily invaded by evil.If the chicken feed contains aflatoxin B1(AFB1),it is easy to damage the spleen and stomach of the chicks,and the spleen and stomach are not properly transported.Transporting and transforming microscopic substances into qi and blood results in reduced disease resistance,suppressed immune function and even death of chicks.Penthorum chinense Pursh compound formula(PCPC)is a compound preparation of Chinese herbal medicine prepared by our laboratory team through clinical syndrome differentiation.PCPC is a mixture of condensed and refluxed volatile oil,water extract,and ultrasonic alcohol extract.In the early stage of the laboratory,PCPC is effective in the treatment of rat liver and gallbladder damp-heat syndrome and the inhibition of Hepg2 liver cancer cells by PCPC,but its mechanism still needs to be To further explore,this experiment explores the protective effect of the compound on the immune function-suppressed chicks caused by AFB1 and the possible mechanism on the spleen.1.To explore the effect of PCPC on AFB1-induced immune function suppression in chicks.By feeding the feed mixed with quantitative AFB1,the immune function suppression model of chicks was established.The group was fed mixed feed containing AFB1,and PCPC or Yinchenhao Decoction was mixed into the AFB1 mixed feed fed every day in the Yinchenhao Decoction and PCPC groups.Chick spleen and thymus organ index,histopathology of chick spleen,and the content of immunoglobulin(Ig A,Ig G,Ig M)in chick serum.The test results showed that after administration of PCPC,it can reduce the morbidity,mortality and mortality of immunosuppressed chickens caused by AFB1;PCPC can significantly down-regulate the increase of spleen index caused by AFB1(P<0.01),reduce the steatosis of the spleen caused by AFB1,The increase of AFB1 resulted in the decrease of serum Ig A,Ig G and Ig M(P<0.01).2.To explore the effect of PCPC on the cytokine levels of TH1/TH2 balance in the spleen of AFB1-induced immune function-suppressed chicks,and the cytokine levels of activated JAK-STAT pathway.The levels of IFN-γ,IL-2,IL-4 and IL-6 in chick spleen were determined by ELISA.Test results: Compared with the blank group,the contents of IFN-γ and IL-2 in the spleen of the model group were significantly decreased(P<0.01),and the contents of IL-4 and IL-6 were significantly increased(P<0.01).Compared with the model group,the content of IFN-γ and IL-2 in the spleen of the chicks in the PCPC medium and low dose groups increased significantly(P<0.01),and the content of IFN-γ in the spleen of the chicks in the high dose PCPC group increased significantly(P<0.01).),the contents of IL-4 and IL-6 in the spleen of chicks in high,medium and low dose groups of PCPC were significantly decreased(P<0.01).Compared with the model group,the contents of IFN-γ and IL-2 in the spleen of the PCPC group were significantly increased(P<0.05),and the contents of IL-4 and IL-6 in the spleen of the PCPC group were significantly decreased(P<0.01).It shows that after adding AFB1 to the feed,the dynamic balance of TH1-TH2 cytokines in the spleen of chicks is broken and shifted to TH2-type cytokines,while PCPC can better alleviate the TH2-type cytokine shift caused by AFB1,and make TH1 cytokines rise again.High,maintain the body’s better immune function.3.To explore the effect of PCPC on the JAK-STAT signaling pathway in the spleen of AFB1-induced immunosuppressed chicks.The m RNA expression levels of JAK-2,JAK-3,STAT-3 and STAT-5 in the spleen were determined by real-time PCR.The test showed that compared with the blank group,the relative expressions of JAK-2,JAK-3,STAT-3 and STAT-5 m RNA in the spleen of chicks in the model group were significantly decreased(P<0.05).Compared with the model group,the relative expressions of JAK-2,JAK-3,STAT-3 and STAT-5 m RNA in the spleen of the PCPC group were significantly increased(P<0.05).4.To explore the effect of PCPC on the levels of apoptotic proteins and anti-apoptotic proteins of AFB1-induced immunosuppression.The protein expressions of Bcl-2,Bcl-xl,Bax and Caspase-3 in the spleen of chicks were determined by ELISA.The test results showed that compared with the blank group,the protein expressions of Bax and Caspase-3 in the spleen of the chicks in the model group were significantly increased(P< 0.01),Bcl-2 and Bcl-xl decreased significantly(P<0.01).Compared with the model group,Bax and Caspase-3 in the spleen of the PCPC group were significantly decreased(P<0.01),and Bcl-2 and Bcl-xl were significantly increased(P<0.01).Ganhuangcao compound can better alleviate the increase of aflatoxin B1-induced spleen apoptotic protein level and the increase of Bcl-2/Bax ratio.In conclusion,this experiment explored the preventive and therapeutic effect of PCPC on AFB1-induced immunosuppressed chicks by adding 3 mg AFB1/kg to the chick feed.Compared with PCPC,it significantly increased the content of Ig A,Ig G,and Ig M in the serum of chicks(P<0.01),increased th1 in the spleen of chicks,decreased the content of th2 cytokines,adjusted the balance of th1/th2 cytokines in the spleen,and increased the jak-2.The relative expression of JAK-3,STAT-3 and STAT-5 m RNA decreased the apoptosis proteins Bax and Caspase-3 in the spleen of chicks(P<0.01),and increased the anti-apoptotic proteins Bcl-2 and Bcl-xl(P<0.01) related.