Therapeutic Effect And Mechanism Of Anemoside B4 On Ovalbumin-induced Allergic Conjunctivitis

Purpose:Evaluate the therapeutic effect of Anemoside B4 eye drops and injections on ovalbumin-induced allergic conjunctivitis,explore its possible mechanism,and provide new medicine and solution for anti-allergic conjunctivitis treatment.Method:Sixty-four male clean-grade SD rats(200-250g in weight)were divided into 8 groups.Except for the normal group,a mixture of 0.02%ovalbumin solution 1ml and 4%aluminum hydroxide adjuvant 1ml was injected intraperitoneally into each rat once every two days in the first 13 days(7 times in total).The mixture contained 40mg aluminum hydroxide,which could slowly release 200?g ovalbumin in vivo to fully sensitize the rats.After 7 times of sensitization,rats were administered according to different treatments of each group for 7 consecutive days.Then 1mol/L DTT 10 ?L was administered in the model group and the administration groups to eliminate conjunctival mucus barrier and improve the effect of ovalbumin challenge.After 15 minutes,the rats were challenged with 2%ovalbumin solution 20 ?L to stimulate allergic conjunctivitis.The therapeutic effect was evaluated by clinical scoring of rat eyes,counting of various stained tissue sections,measurement of changes in vascular permeability by injection of Evans Blue in the tail vein,and determining the proportion of T cell subsets in rat splenocytes by flow cytometry.The possible mechanism was explored by measuring the content of cytokines in serum by ELISA,measuring the content of IgE in serum by ELISA,determining the proportion and quantity of peripheral blood leukocyte by automatic blood cell analyzer,and determining of protein expression differences in splenocytes by Western Blot.Results:In ovalbumin-induced allergic conjunctivitis,Anemoside B4 reduced the signs of ocular inflammation and clinical scores,inhibited the structural changes of conjunctiva in tissue sections,reduced infiltration of eosinophils in conjunctiva,and inhibited degranulation of mast cells.It reduced the amount of Evans Blue exudation,improved the vascular permeability of rat eyes,recovered the ratio of CD4+T cells to CD8+T cells,and recovered the internal immune environment.Anemoside B4 in Western Blot inhibited the expression of p-JNK and p-p38.The mechanism of cytokines,IgE and peripheral blood leukocytes remained to be clarified in future studies.Conclusion:Anemoside B4 has a clear effect on the treatment of ocular changes caused by allergic conjunctivitis.The research results can provide new options and references for the development of new medicine for the treatment of allergic conjunctivitis and related eye diseases.It can lay an important foundation for the successful development of a new generation of anti-allergic conjunctivitis medicines,and can provide new ideas for the treatment of allergic eye diseases if further research on its effects and mechanism can be implemented.