雄激素替代对兔混合性干眼的治疗效果

Effect of androgen replacement therapy on mixed dry eye in a rabbit model

目的探索雄激素替代对合并雄激素水平低下和睑板腺功能障碍的兔干眼模型的治疗效果。方法将6月龄雄性青紫蓝兔计算机随机分为治疗组、造模组及对照组各10只。治疗组及造模组利用电凝笔烧灼封闭2/3睑板腺开口,并切除双侧睾丸。造模28 d起治疗组在两侧眶周皮肤涂抹1%睾酮凝胶1 g/d,造模组及对照组涂抹凡士林,共治疗28 d。监测泪液分泌、泪膜破裂时间(TBUT)、角膜荧光染色及血清游离睾酮。实验终点取眼球及眼睑进行苏木精-伊红(HE)染色和过碘酸希夫(PAS)染色,测量结膜miRNA-744-5p和白细胞介素6(IL-6)的表达量并检测睑脂脂肪酸成分。结果造模28 d后兔出现典型干眼体征及雄激素缺乏。雄激素替代治疗28 d后,与造模组相比,治疗组泪液分泌[(14.7±5.2)比(10.3±3.6)mm,P=0.001]、TBUT[(15.0±4.2)比(10.2±3.6)s,P=0.003]及角膜荧光染色评分[0(0,1)比2(1,4)分,P<0.001]均改善,穹窿杯状细胞数量增加[(27.2±7.6)比(10.7±4.8)个,P<0.001]。与造模组相比,治疗组结膜miRNA-744-5p(1.67±0.24比2.63±0.58,P<0.001)及IL-6[2.38(1.84,4.61)比4.82(3.99,6.36),P=0.022]的表达水平均降低,睑脂16∶1,Δ^(9)脂肪酸[(10.31±1.00)%比(3.87±0.45)%,P<0.001]及iso-18∶0脂肪酸[(7.09±0.93)%比(2.44±0.70)%,P<0.001]相对含量增加,iso-26∶0[(5.72±1.07)%比(8.02±0.65)%,P<0.001]相对含量降低。结论雄激素替代能够改善合并雄激素水平低下及睑板腺功能障碍的兔干眼治疗效果。

Objective To explore the effect of androgen replacement therapy in a rabbit dry eye model characterized by androgen deficiency and meibomian gland dysfunction(MGD).Methods Thirty 6-month-old male Chinchilla rabbits were randomly divided into the treatment group,model group and control group,with 10 rabbits in each group.In the treatment and model groups,2/3 of the meibomian gland openings were closed by cauterization with electric coagulation pen,and bilateral testes were removed.One gram gel containing 1% testosterone was applied for 28 days on the skin of rabbits in the treatment group since day 28 after the surgery.The model group and control group received transdermal petrolatum instead.Tear secretion,tear breakup time(TBUT),corneal fluorescein staining,and serum free testosterone level were monitored throughout the study period.The globes and eyelids were collected for hematoxylin-eosin staining and periodic acid-Schiff staining.Conjunctival tissue was tested for the expression of miRNA-744-5p and interleukin-6.Meibum was collected for fatty acid analysis.Results Animals presented with typical dry eye signs and androgen deficiency.After 28-day androgen replacement therapy,compared with the model group,the treatment group had a significantly higher tear secretion rate[(14.7±5.2)vs(10.3±3.6)mm,P=0.001],higher TBUT[(15.0±4.2)vs(10.2±3.6)s,P=0.003],lower fluorescein staining score[0(0,1)vs 2(1,4),P<0.001],and higher goblet cell density at conjunctival fornix(27.2±7.6 vs 10.7±4.8,P<0.001).Additionally,compared with the model group,the conjunctiva of the treatment group expressed a significantly lower level of miRNA-744-5p(1.67±0.24 vs 2.63±0.58,P<0.001)and interleukin-6[2.38(1.84,4.61)vs 4.82(3.99,6.36),P=0.022].Meanwhile,the treatment group showed significantly increased level of 16∶1,Δ^(9) fatty acid[(10.31±1.00)%vs(3.87±0.45)%,P<0.001]and iso-18∶0 fatty acid[(7.09±0.93)%vs(2.44±0.70)%,P<0.001],but decreased level of iso -26∶0 fatty acid[(5.72±1.07)%vs(8.02±0.65)%,P<0.001]in the meibum compared with the model group.Conclusion Androgen replacement therapy can alleviate dry eye signs in rabbits presented with combined androgen deficiency and MGD.