Effect And Mechanism Of Cordy Cepinon Induced Inflammation Of Paw By CFA Adjuvant In Mice

Objective: Preliminary Studies on the anti-inflammatory mechanism of cordycepin in adjuvant-induced inflammation in mice.Methods: Sixty female KM mice were constructed as models and divided in 6 groups: : control group(Control),saline group(Model),cordycepin high(H-Cor)dose group(20 mg/kg),cordycepin mid(M-Cor)dose group(10 mg/kg),cordycepin low(L-Cor)dose group(5 mg/kg),dexamethasone positive control group(Dexa).Mice in Cordycepin group were injected with different doses of cordycepin solution;mice in positive group were treated with dexamethasone(2mg/kg);the blank control group and physiological saline group were given the same dose of normal saline.Each mouse was administered by intraperitoneal injection once daily for 7 days.after the last administration of1 h,mice were induced to swell with CFA to establish an inflammation model.Photographs were taken at 4 h,8 h,12 h,24 h,48 h,;after the model was established to measure the thickness of the right hind paw and observe the changes in the degree of swelling of 72 h.The spleen and thymus index of the mice were measured and calculated,and photographs were recorded;HE staining was performed.Pathological changes of the inflammatory paws of mice;mouse protein antibody arrays to detect the trend of changes in cytokines at the site of inflammation;selection of changes in the levels of related inflammatory factors;detection of TNF-a,IL-1β,IL-6,IFN in serum by ELISA The content of cytokines such as γ was changed;the density of macrophages(F4/80)and T cells(CD3+)in inflammation sites was marked by immunohistochemistry,and the expression levels of relevant cytokines secreted by the two immune cells were verified.Result:1.Changes in paw swelling of mice induced by inflammation From the results of the photo,it can be observed that there was no significant difference in paw swelling changes of the Control group mice at6 points after0h;compared with the Control group,the paw swelling of the Model group significantly increased;after the model was replicated,the foot swelling reached its peak at 12 hours.the degree of swelling was still obvious after 72 h.Compared with 0 h,after the model was replicated,the paws of 12 h mice were not obvious in H-Cor,M-Cor-treated and Dexa-treated mice,and the degree of swelling gradually subsided after 72 h;There was no significant difference in the L-Cor treatment group compared to the Saline treatment group.2.Changes of immune organ index in miceCompared with mice in the Control group,the spleen and thymus in the Saline-treated group were significantly enlarged,and the spleen and thymus were significantly increased(P<0.05);however,compared with the Saline-treated group,the splenomegaly in M-Cor and Dexa treatment group was inhibited and the spleen index significantly decreased.(P<0.05).There was no significant difference between the H-Cor and L-Cor groups and the Model group.Compared with Model group,the thymus index of Cor treatment group andDexa treatment group also decreased significantly(P<0.05),and there was no significant difference in the reduction of thymus index between Cor treatment groups and Control group;thymus and spleen of Dexa treatment group.There is no significant difference between the index and the Control group.3.Pathological changes in the paw of mice inducedby inflammation In the control group,the mouse foot was normal,but the paw of the Model group was significantly damaged.Lymphocytes,monocytes,and a few infiltrated neutrophils were seen infiltrated;the M-Cor treatment group and the Dexa treatment group were compared with the Model group.The structure of the paw was clear,the degree of injury was light,and there was no obvious infiltration of inflammatory cells.The soft tissue structure of the H-Cor treated group was slightly damaged.There was no significant difference between the L-Cor treated group and the Model group.4.Trend of cytokines in inflammation sites in miceBy using a mouse cytokine antibody chip to detect the tendency of cytokines in mouse inflammatory tissues,we found that compared with Control,MMP-1,IL-1,IL-3,IL-4,and IL-6 were observed in the Model group.IL-12,IL-13,IL-17,TNF-α,BLC,CCL11,MIP-1,etc.all have a tendency to increase;but compared with Model,M-Cor and Dexa treatment groups not only significantly inhibited the increase of proinflammatory cytokines such as IL-3,IL-2,IL-6,IL-9,IL-13,TNF-α,etc.but also suppressed the inflammatory chemokines MCP-1,BLC,CD30,CCL11,MIP-α,etc.;compared with M-Cor,the inhibitory effects of H-Cor and L-Cor treatment groups were not significant.5.verify the expression of some cytokines in mouse serum by Elisa method Compared with the Control group,the serum levels of TNF-α,IL-6,IL-1β,and IFN-γ in the Saline-treated group significantly increased(P<0.05);comparedwith the Saline-treated group,H-Cor,M The contents of TNF-α,IL-6,IL-1β and IFN-γ in the Cor-and Dexa-treated groups were significantly lower(P<0.05).The M-Cor-treated group was significantly lower than the H-Cor-treated group(P< 0.05);Compared with Model group,there was no significant difference in L-Cor treatment group;there was no significant difference between M-Cor and Dexa treatment group.6.Detection of mouse immune cells and their related cytokine expressionCompared with the Control group,the number of macrophages(F4/80)and T lymphocytes in the Model group and L-Cor-treated group was significantly higher(P<0.05),and there was no significant difference in the number of positive cells between the two groups.Compared with Model group,H-Cor treatment group,M-Cor treatment group and Dexa treatment group can significantly reduce the number of macrophages(F4/80)and T lymphocytes in inflammatory tissues(P<0.05).There was no significant difference in the positive rate of macrophages between the H-Cor and Dexa treated groups compared with the Control group.Conclusion: Cordycepin can significantly improve the pathological response of inflammation in mice,reduce inflammatory swelling and pathological injury of the soles of the feet,and reduce the immune organ index.The mechanism may be that cordycepin can inhibit the secretion of pro-inflammatory cytokines such as TNF-α,IL-1β,IL-6,IFN-γ in the blood,and may inhibit the inflammatory chemokines at the inflammation sites(MCP-1,The release of BLC,CD30,CCL11,MIP-α,etc.,and the reduction in the number or aggregation of macrophages,T cells,and other epidemic cells eventually inhibit the expression of related inflammatory cytokines in inflammation sites and improve adjuvant-induced inflammatory responses.Inaddition,the inflammation model in this study is similar to the international model for adjuvant arthritis.Therefore,cordycepin may play a therapeutic role in inflammatory diseases such as adjuvant arthritis.