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目的观察密蒙花颗粒剂对去势雄兔泪腺细胞凋亡因子Bax、Bcl-2、Fas和Fas L的影响,探讨密蒙花颗粒剂对去势雄兔干眼症模型的疗效。方法将密蒙花原药材制备成颗粒剂。将30只健康成年新西兰长耳白兔(雄性),随机分为空白组(A组),模型组(B组),密蒙花颗粒剂组(C组),安慰剂组(D组),睾酮组(E组)5组,每组6只。除A组外其余4组实验用兔行双侧睾丸及附睾切除术。除A、B组外,从术后第3日开始C组予密蒙花颗粒剂,按100 mg/kg灌胃,3次/日;D组予生理盐水20 m L灌胃,3次/日;E组按2 mg/kg在大腿肌肉处注射丙酸睾酮注射液,每3日注射1次。各组实验用兔,分别于术前和术后第4周各进行1次SchirmerⅠ试验(SchirmerⅠtest,SⅠT)和泪膜破裂时间(tear break-up time,BUT)测定。用药观察4周后采用空气栓塞法处死各组兔子,摘取双眼泪腺组织,采用免疫组化法对泪腺细胞凋亡因子Bax、Bcl-2、Fas和Fas L的表达进行检测,并观察泪腺组织结构形态图。结果 (1)SⅠT及BUT结果:与本组术前及术后A组比较,B、D组SⅠT及BUT均明显下降(P<0.01);与B、D组术后比较,C、E组SⅠT及BUT升高,且差异均有统计学意义(P<0.01)。(2)Bax、Bcl-2、Fas、Fas L表达:与A组比较,B、D组Bax、Fas、Fas L表达均明显增多,Bcl-2表达明显减少(P<0.01);与B、D组比较,C、E组Bax、Fas、Fas L表达均明显减少,Bcl-2表达明显增多(P<0.01)。(3)Bax、Bcl-2、Fas、Fas L表达观察结果:A组:泪腺结构清晰,未见Bax、Bcl-2、Fas、Fas L表达;B组:泪腺结构模糊,Bax、Fas、Fas L均大量表达于细胞膜和细胞浆中,呈棕黄色颗粒,Bcl-2仅少量表达;C组:泪腺结构清晰,散见Bax、Fas、Fas L表达,Bcl-2大量表达于细胞膜和细胞浆中,呈棕黄色颗粒;D组:泪腺结构模糊,Bax、Fas、Fas L均大量表达于细胞膜和细胞浆中,呈棕黄色颗粒,Bcl-2仅少量表达;E组:泪腺结构清晰,散见Bax、Fas、Fas L表达,Bcl-2未见表达。结论密蒙花颗粒剂具有与雄激素相似的抑制泪腺细胞凋亡因子Bax、Fas和Fas L表达,同时上调Bcl-2的作用,能抑制雄激素缺乏所致兔泪腺细胞的凋亡,维持泪腺基础分泌量,但其作用弱于雄激素。
Objective To observe the effects of granulation granule on the apoptosis factors Bax, Bcl-2, Fas and Fas L of ovarian lacrimal glands in ovariectomized rabbits and to explore the curative effect of granule of Mongolianum Mung on the model of dry eye of castrated male rabbits. Methods To prepare the granules of Budweiser medicinal. Thirty healthy adult New Zealand white rabbits (male) were randomly divided into blank group (group A), model group (group B), granule group (group C), placebo group Testosterone group (E group) 5 groups, 6 in each group. Except group A, the remaining four groups of rabbits with bilateral testis and epididymal resection. In addition to group A, B, from the 3rd day after the operation, the patients in group C were treated with granule of Mongolianum granulosum at the dose of 100 mg / kg orally 3 times daily; group D was given saline 20 ml once daily for 3 times / Day; E group by 2 mg / kg in the thigh muscle injection of testosterone propionate injection, once every 3 days. Rabbits in each group were subjected to 1 Schirmer I test (Schirmer I test, STI) and tear break-up time (BUT) measurements at preoperative and 4th weeks respectively. After rabbits were sacrificed by air embolism for 4 weeks, rabbits in each group were sacrificed and their tear lacrimal glands were removed. Immunohistochemistry was used to detect the expression of Bax, Bcl-2, Fas and Fas L cells in lacrimal gland, Structure diagram. Results (1) SⅠT and BUT results: Compared with preoperative and postoperative group A, SⅠT and BUT in group B and D were significantly decreased (P <0.01); Compared with group B and D, group C and E S Ⅰ T and BUT increased, and the differences were statistically significant (P <0.01). (2) The expression of Bax, Bcl-2, Fas and Fas L in B and D group were significantly increased compared with A group (P <0.01) Compared with group D, the expression of Bax, Fas, Fas L in group C and group E were significantly decreased and the expression of Bcl-2 was significantly increased (P <0.01). (3) The expression of Bax, Bcl-2, Fas and Fas L: Group A: The structure of lacrimal gland was clear, but Bax, Bcl-2, Fas and Fas L were not expressed; L were expressed mainly in the cell membrane and cytoplasm, showing brown granules, only a small amount of Bcl-2 expression; C group: clear structure of the lacrimal gland scattered Bax, Fas, Fas L expression, Bcl-2 large expression in the cell membrane and cytoplasm D group: the structure of lacrimal gland was obscure, Bax, Fas and Fas L were abundantly expressed in the cell membrane and cytoplasm, with brownish yellow granules, while only a small amount of Bcl-2 was expressed; Group E: See Bax, Fas, Fas L expression, Bcl-2 was not expressed. Conclusions Mumeng Hua Granule can inhibit the expression of apoptosis factors Bax, Fas and Fas L of lacrimal gland cells in a similar way to androgen, and up-regulate the expression of Bcl-2, and can inhibit the apoptosis of lacrimal gland cells in androgen-deficient rabbits. Basal secretion, but its role is weaker than androgen.