雄激素调节小鼠泪膜功能与角膜上皮细胞Mucins表达的体内研究

Effect of androgen on mice tear film stability and Mucins expressions in mice corneal epithelial cells in vivo

目的:探讨去势小鼠干眼模型中雄激素浓度对小鼠泪膜稳定性以及角膜上皮细胞Mucins表达的影响。方法:通过双侧睾丸切除去势手术,改变体内雄激素水平,观察泪膜破裂时间的变化,同时在不同时间点采用RT-PCR法与Western-Blot法分别检测小鼠角膜上皮细胞中Muc1和Muc4的mRNA和蛋白水平的表达变化。结果:去势1wk后小鼠睾酮质量浓度下降至0ng/μL;正常对照组、伪手术组及去势术后1、2、4、6、8wk组小鼠BUT分别为68.33±12.86、62.47±3.75、58.67±10.03、47.17±7.64、39.51±3.39、32.67±3.88、31.00±2.36s,其中术后2、4、6、8wk组小鼠泪液BUT值较正常对照与伪手术组均明显缩短,差异均有统计学意义(均P<0.05)。Muc1与Muc4的mRNA与蛋白水平表达,随雄激素浓度下降,表达下调(P<0.05),Muc1在去势后第2wk表达最低,而Muc4在去势后第1wk表达降至最低。结论:在体内,雄激素可调控小鼠角膜上皮Mucins表达,并缩短泪膜破裂时间,参与泪膜稳定性的维持。

AIM： To study in vivo the effect of androgen on mice tear film stability and Mucins expressions in corneal epithelial cells in BALB/ c mice after orchectomy. METHODS： With orchiectomy operation, we set up mice model. And serum androgen concentration of mice was detected by radioimmunoassay. Break - up time （ BUT ） of tear film was tested in the different experimental points. Mice corneal epithelia were peeled and MUC1 and MUC4 mRNA and protein levels were measured by RT-PCR and Western blot. RESULTS： The serum androgen concentration reduced to 0ng/ μ L at 1wk after orchiectomy. The BUT values were 68. 33±12. 86s, 62. 47±3. 75s, 58. 67± 10. 03s, 47. 17±7. 64s, 39. 51±3. 39s, 32. 67±3. 88s and 31. 00±2. 36s in the normal control group, sham group and in orchectomy group at 1, 2, 4, 6 and 8wk, respectively, and the BUTs＆amp;nbsp;were significantly shorter in the orchectomy at 2, 4, 6 and 8wk groups than those in the normal control group （all at P〈 0. 05）. MUC1 and MUC4 mRNA and proteins levels decreased with androgen level lowering （P〈0. 05）. Mucin1 level was the lowest at 2wk after orchiectomy, and the lowest Mucin4 level was found at 1wk after orchiectomy. CONCLUSION： In vivo, androgen regulates Mucins expressions in mice corneal epithelial cells, makes BUT shorter,and influence the stability of tear film.