Effects Of Butterflybush Flower Eye Drops On Expression Of Inflammatory Cytokines IL-1β,Mucin 5AC And P38MAPK In Castrated Male Rabbits

Object:To observe the effects of butterflybush flower eye drops at different concentrations on expression of inflammatory crtokines IL-1β,Mucin 5AC and P38 MAPK in castrated male rabbits.Method : Thirty-six male rabbits were randomly divided into blank group(A),model group(B),low concentrations butterflybush flower eye drops group(C,1 mg/ml),medium concentrations butterflybush flower eye drops group(D,1.5 mg/ml),and high concentrations butterflybush flower eye drops group(E,3 mg)./ml)concentration group,and testosterone group(F).Except for group A,bilateral testes and epididymis were excised.8 weeks after the operation,ocular surface examination showed that the model was successful.Group A and B were not treated.Group C,D and E were given different concentration of butterflybush flower eye drops 3 times per day,and the F group according to 0.5 ml/kg dose in the thigh muscles injected with testosterone propionate,once every 3 days.All the rabbits in each group were measured with fluorescein staining,Schirmer I test(SIT)and tear break-up time(BUT)respectively before and after 4 weeks of medication.Four weeks after treatment,rabbits were killed and their conjunctival tissues were collected.Immunohistochemical staining was used to detect the expression of IL-1β,Mucin 5AC and P38 MAPK in the conjunctiva and statistical analysis was performed.Results:1.SIT,BUT,and corneal fluorescein staining: there was no significant change in the length of tear secretion filter,rupture time of tear film and fluorescein stain in group A(P>0.05).In group B,the length of the tear secretion was shortened,the tear film rupture time was significantly lower than that of other groups,and the corneal fluorescein staining was positive,with significant difference(P<0.01).C,D,and E group were compared with group B,and we can find the length of the tear secretion was prolonged,the tear time was prolonged,the corneal fluorescein staining was positive,and the difference was significant(P<0.01).The length of the tear filter in the group D and E was longer than that in the group C,and the tear film rupture time was higher than that in the group C.The difference was statistically significant(P<0.01).2.Results of IL-1β expression in conjunctiva by immunohistochemical staining: In group B,there was a large number of inflammatory cell infiltration in the conjunctiva,and IL-1β had a strong positive expression.There is astatistically significant on the differences between group B and group A,C,D,E,and F(P<0.01).In group C,D,E,inflammatory cells were significantly decreased compared with group B,but more than group E,and there was a significant difference compared with group E(P<0.05).The difference between group C and group D and group E was statistically significant(P<0.05).There was no significant difference between group D and group E(P>0.05).3.Results of Mucin 5AC expression by conjunctival immunohistochemistry:A small amount of MUC5 AC was found in the conjunctiva of group B after modeling,which was significantly different from that in group A,C,D,E,F(P<0.01).After treatment,the expression of MUC5 AC in group C,D and E was significantly higher than that in group B.Compared with group D and group E,the expression of MUC5 AC in group C was lower than that in group D and E,and the difference was statistically significant(P<0.05).There was no significant difference between D group and E group(P>0.05). There was no significant difference between group D and group E and group F(P>0.05).However,the secretion of MUC5 AC in group F was more than that in groups D and E.4.Results of P38 MAPK expression of conjunctivitis immunohistochemistry:compared with A,positive expression of P38 MAPK in group B was significantly increased(P<0.01).The expressions of group B and group C,D,E and F were significantly reduced,and the difference was statistically significant(P< 0.01).Compared with group D,group E and F,the expression of P38 MAPK in group C was increased with statistical significance(P<0.01).There was no significant increase in the positive expression rate of group D and group E,and the difference was not statistically significant(P>0.05).There was significant difference between D group and E group and F group(P<0.01).Conclusion:With bilateral orchiectomy,different concentrations of butterflybush flower eye drops have androgen-like effects,which can increase the basal secretion of tears and stabilize tear film function in castrated male rabbits after dry eye treatment.Different concentrations of butterflybush flower eye drops can reduce the expression of interleukin-1βin the conjunctiva of dry eye and increase the expression of Mucin 5AC and P38 MAPK in the conjunctiva of dry eye,thus decreasing the inflammatory infiltration of dry eye conjunctiva and increasing the dry eye mucin protein.The expression maintains tear film stability,but its effect is weaker than androgens.The effects of medium and high concentrations of butterflybush flower eye drops were stronger than those of low concentrations,while the high concentration group is superior to the medium concentration group.