菊花总黄酮对去势导致雄兔干眼症泪腺细胞Bax、Bcl-2表达的影响

Effects of Total Flavonoids of Chrysanthemum on Bax and Bcl-2 Expressions of Lacrimal Gland Cells Apoptosis of Castrated Rabbits with Dry Eye

目的观察菊花总黄酮对去势所致干眼症白兔泪腺凋亡相关基因蛋白Bax、Bcl-2表达的影响,探讨菊花总黄酮治疗干眼症的作用机制。方法将150只雄性日本大耳白兔随机分为正常组(A组)、假手术组(B组)、模型组(C组)、雄激素对照治疗组(D组)和菊花总黄酮治疗组(E组)。分别于治疗后1、3和5个月每组处死10只,取材用于相关指标检测。C组、D组、E组行双侧去势术建立白兔干眼症模型。E组白兔用菊花总黄酮灌胃治疗,D组用雄激素肌肉注射,A组、B组、C组用生理盐水灌胃。全部白兔行SchirmerⅠ实验,并检测泪膜破裂时间,处死后取泪腺组织,采用免疫组织化学法检测泪腺组织中凋亡相关基因蛋白Bax、Bcl-2的表达,并计数凋亡细胞数量。结果 E组SchirmerⅠ实验测量所得滤纸湿长明显高于C组(P<0.01),泪膜破裂时间明显长于C组(P<0.01)。E组治疗1、3和5个月后,泪腺导管及腺泡上皮细胞中Bax阳性表达的细胞数均明显低于C组,Bcl-2阳性表达的细胞数均明显高于C组,细胞凋亡数量均明显低于C组(P<0.01)。结论菊花总黄酮中主要成分为黄酮类物质,可显著抑制雄激素水平降低后白兔干眼症的发生,抑制泪腺细胞凋亡,维持泪腺基础分泌量和泪膜的稳定性。

Objective To evaluate the effects of the total flavonoids of chrysanthemum on basic tears secretory volume, tear film stability, expressions of Bax and Bcl-2 of castrated rabbits with dry eye, and investigate the therapeutic effects of the total flavonoids on dry eye caused by gonadal hormones level imbalance. Methods 150 rabbits were divided into blank group （group A）, sham-operated group （group B）, untreated group （group C）, androgen group （control treatment group, group D）, and total flavonoids of chrysanthemum group （treatment group, group E）. The dry eye models were established with orchiectomy on the group C, group D and group E. Rabbits in the group E were treated with total flavonoids of chrysanthemum. Rabbits in the group D were treated with androgen intramuscular injection. Rabbits in the group A, group B and group C were treated with normal saline. All rabbits were detected with Schirmer Ⅰ experiment and the tear break-up time （BUT） was evaluated for dry eye. Bax, Bcl-2 were checked on immunohistochemistry. Results The Schirmer＇s Ⅰ test values of the treatment group was significantly higher than that of the untreated group （P〈0.01） and the BUT value of the treatment group was significantly longer than that of the untreated group （P〈0.01）. The quantity of expression of Bax in acinar epithelial cell and glandular tube cell of the treatment group were significantly higher than that of the untreated group （P〈0.01） and the expression of Bcl-2 in acinar cell and glandular tube cell of the treatment group were also significantly higher than that of the untreated group （P〈0.01）. Conclusion The main components of chrysanthemum are the flavonoids, which could significantly inhibit the syndrome of dry eye in rabbit after androgen level lowered and lacrimal gland apoptosis and keep basic tears secretory volume and tear film stability.