The Regulatory Effects And Mechanisms Of Flavonoids From Chromolaena Odorata On Immunity In Mice

Chromolaene odorata Linn.is a perennial herb or subshrub plant in the Asteraceae family.Itoriginated from Central America,and is the first invasive plant in China.It is mainly distributed in the southern area of China.C.odorata is commonly used as a medicine by folks to cure colds,diarrhea,and skin diseases.The main component of the plant is flavonoids,which have various biological activities.In this study,the aerial part of the plant was collected to extract flavonoids by ultrasonic-assisted ethanol extraction method.the regulartory effects and mechanisms of flavonoids from C.odorata?FCO?on immunity in Kunming mice was investigated.This provides experimental basis for the development of new Chinese herbal feed additives.Several tests below were conducted in this study:?1?FCO was extracted using 70%ethanol as a solvent,and assisted by ultrasonic technology,with material-liquid ratio of1:50?g/m L?,extraction temperature of 70°C,extraction time of 6 h.The flavonoids components were detected by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry?UHPLC-QTOF-MS?technology after silica gel column purification.?2?Kunming mice were used to establish immunosuppressed model by cyclophosphamide to investigate the effects of different dosage(30,150 and 750 mg·kg^-1·d)of FCO on immune organs?thymus and spleen?indices,blood parameters?WBC,LYM,MON,NEU,ESO,BASO,RBC,HGB and PLT?,serum immunoglobulins?Ig G and Ig M?,serum cytokines?IFN-?,IL-2 and TNF-??,and tissue structure of immune organs.?3??3?RT-q PCR technology was used to study the effect of FCO on the m RNA relative expression of IFN-?,GATA3 and Tbx21 of spleen in mice.The results were found as follows:?1?The extraction rate of FCO was 8.29%.38kinds of flavonoids were detected by UHPLC-QTOF-MS.?2?The thymus index and spleen index in each group significantly decreased?P<0.01?compared to blank control group.The immune organs indices of FCO high-dose group significantly increased compared with immunosuppressed group?P<0.05?.Results of hematology tests showed that compared with immunosuppressed group,WBC of FCO test groups,LYM and MON of FCO high-dose group and NEU of FCO medium-dose group significantly increased?P<0.05?.There was no significant difference in EOS and BASO in each group?P>0.05?.RBC and HGB of FCO test groups,PLT of FCO high-dose group significantly increased compared with those in immunosuppressed group?P<0.05?.Results of serum immunoglobulin showed that Ig G of FCO high-dose group and Ig M of FCO medium-dose,and high-dose groups significantly increased compared with immunosuppressed group?P?0.05?.Results of serum cytokine test showed that though there was no significant difference of IFN-?of FCO test groups compared to immunosuppressed group.There was an increasing trend among these three groups.No significant difference was seen in IL-2 among each group?P>0.05?TNF-?of FCO medium-dose and high-dose groups significantly increased compared to immunosuppressed group?P<0.01?.Results of pathology tests showed that the number of cells were normal and the structures were intact of thymus and spleen in blank control group,while in the immunosuppressed group the cells were heavily reduced and the structures were severely damaged.The number of cells decreased slightly and some of the damaged structures recovered on varying degrees of immune organs in the FCO test groups.?3?Compared with immunosuppressed group,the m RNA relative expression of Tbx21 and IFN-?of spleen in FCO test groups significantly increased?P?0.05?,and the m RNA relative expression of GATA3 significantly decreased?P?0.05?.In conclusion,the extraction rate of FCO by ultrasonic-assisted ethanol extraction method was high.A certain dose of FCO increased the immune organ indices,the levels of WBC,LYM,MON,NEU,RBC,HGB and PLT,the contents of Ig M,Ig G and TNF-?and recovered the damaged structures and maintained the number of cells of the immune organs of immunosuppressed mice.FCO at a certain dose increased the m RNA relative expression of IFN-?and Tbx-21 and decreased the m RNA relative expression of GATA3,thereby improving the imbalance of the Th1/Th2 of the body,thereby ensuring immune regulation.