The Expression Of CC Chemokine Ligand 20 In Ocular Surface With Dry Eye Mice

Objective:To investigate the role of CC chemokine ligand 20(CCL20) in the pathogenesis of dry eye. by evaluating CCL20 expression in the ocular surface and the dry eye inhibit role of CCL20 neutralization.Methods:Eighteen female C57BL/6 mice,six-week old, were randomly divided into three groups equally: the normal control group,the dry eye group and the anti-CCL20 neutralization antibody treatment group. Control group were raised in normal room enviroment. without interevention Dry eye mode was established by double lower limbs subcutaneous injection scopolamine hydrobromide at four fixed time points(8:30,11:30,14:30,17:30) combine constant mid-speed airflow.in a environment control chamber(temperature between 22 to 24, relative humidity below 17%).Both dry eye group and anti-CCL20 neutralization antibody treatment group mice were induced dry eye mode and separately treated with 1% PBS and anti-CCL20 neutralization antibody by subconjunctival injection 2 days before inducing dry eye and at days 2,4,6,8,10 after inducing dry eye. Corneal fluorescein staining(CFS) was used to assess dry eye severity Collect tears,cornea epithelium, conjunctiva tissue of each group and detect the the expression of CCL20 and(or) IL-17 by Real-Time PCR, immunohistochemical staining and luminex liquichip.Results:1. The cornea CCL20 m RNA, IL-17 m RNA expression of normal control group,dry eye group and the anti-CCL20 antibody treatment group:1.00±0.00, 1.45±0.11,0.23±0.04; 1.00±0.00, 4.05±0.19, 1.67±0.34. The conjunctiva CCL20 m RNA, IL-17 m RNA expression of normal control group,dry eye group and the anti-CCL20 antibody treatment group:1.00±0.00、6.38±1.86、0.32±0.14; 1.00±0.00、4.08±1.38, 0.74±0.29.CCL20 m RNA, IL-17 m RNA expression in cornea and conjunctiva of dry eye group mice increased significantly compared with normal control group and anti-CCL20 neutralization antibody treatment group with statistically significant(P<0.05).2. The CCL20, IL-17 tear concentration of normal control group, dry eye group and the anti-CCL20 antibody treatment group:(50.39±7.65) pg/ml,(134.45±7.0)pg/ml,(17.52±5.83)pg/ml;(100.9±10.12)pg/ml,(220.88±19.15)pg/ml,(64.11±5.56)pg/ml.T he tear concentration of CCL20 and IL-17 showed significant difference among 3 groups analys is of variance(P<0.05).3. Through immunohistochemical staining by anti-CCL20 antibody, brown particles can be observed in the ocular surface of dry eye group mouse while not seen in normal control group and the anti-CCL20 antibody treatment group.Conclusion:CCL20 involved in the pathogenesis of dry eye, subconjunctival injection anti-CCL20 antibody can inhibited the expression of CCL20, as well as another important inflammatory cytokine IL-17,relieve dry eye ocular surface damage.