Effects of Ophiopogonanone A on Proliferation of Leukocytes and Goblet Cells in Conjunctiva of Mice with Dry Eye Disease
Objective:To explore the therapeutic effect and mechanism of Ophiopogonanone A(OPA)on dry eye syndrome(DED)mice.Methods:Mice were divided into five groups:NC group(n=12),DED group(n=14),L-OPA group(n=14),M-OPA group(n=14)and H-OPA group(n=14).The NC group was normal control mice,and the other groups were mice with dry eye disease induced by benzalkonium chloride.Mice in NC group and DED group were intraperitoneally injected with 1％dimethyl sulfoxide(DMSO),while mice in L-OPA group,M-OPA group and H-OPA group were intraperitoneally injected with OPA of 10,20 and 40 mg/kg/d,respectively.The rats were treated for 4 weeks.After treatment,the tear secretion of each group was measured,and corneal fluorescein sodium staining,conjunctival Periodic Acid-Schiff stain(PAS)and corneal TUNEL staining were performed.The levels of interleukin-1β(IL-1β),intercellular adhesion molecule-1(ICAM-1)and tumor necrosis factor-α(TNF-α)in tears were measured by ELISA method.The levels of Bax and Bcl-2 mRNA in cornea were measured by RT-qPCR.The protein expression of conjunctival K10 and leukocyte common antigen(LCA)was detected by Western blot or immunohistochemical staining.Results:Compared with NC group,tear secretion and tear break-up time(BUT)decreased(P＜0.05),corneal fluorescein sodium staining grade increased(P＜0.05),tear IL-1β,ICAM-1 and TNF-α levels increased(P＜0.05),the number of conjunctival goblet cells decreased(P＜0.05),corneal TUNEL positive rate and Bax mRNA relative expression increased(P＜0.05),corneal Bcl-2 mRNA relative expression decreased(P＜0.05),corneal K10 protein relative expression increased and conjunctival LCA protein relative expression increased(P＜0.05).Compared with DED group,tear secretion and BUT in L-OPA,M-OPA and H-OPA groups increased(P＜0.05),corneal fluorescein sodium staining grade decreased(P＜0.05),tear IL-1β,ICAM-1 and TNF-α levels decreased(P＜0.05),conjunctival goblet cell number increased(P＜0.05),corneal TUNEL positive rate and Bax mRNA relative expression decreased(P＜0.05),corneal Bcl-2 mRNA relative expression increased(P＜0.05),corneal K10 protein relative expression decreased and conjunctival LCA protein relative expression decreased(P＜0.05).Conclusion:Ophiopogonanone A is effective in the treatment of benzalkonium chloride-induced dry eye disease in mice,and its mechanism may be related to the inhibition of leukocyte-mediated inflammation and the promotion of goblet cell proliferation.
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