贵州医科大学学报

2020, v.45;No.237(06) 649-654

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吉法酯保护干眼症大鼠的机制研究
Study of Protective Mechanism of Gefarnate on Dry Eye in Rats

糜玲;周波;李永;彭金刚;汤磊;黄家宇;
MI Ling;ZHOU Bo;LI Yong;PENG Jingang;TANG Lei;HUANG Jiayu;Department of Pharmaceutical Analysis,Guizhou Medical University;Guizhou Province Chemical Synthetic Drug Research and Development Engineering Technology Research Center;

摘要(Abstract):

目的:探讨不同构型吉法酯对干眼症大鼠的保护机制。方法:Wistar雌鼠40只,8只大鼠不予任何处理作为正常组,32只通过连续28 d皮下注射东莨菪碱氢溴酸盐并联合高流通、低湿度环境下制备干眼症模型后均分为生理盐水组、4Z吉法酯治疗组(4Z组)、4E吉法酯治疗组(4E组)及混合吉法酯治疗组(4Z∶4E=30∶70,混合组),治疗14 d后将大鼠冰上断颈处死,采用免疫组织化学染色检测各组大鼠眼表基质金属蛋白酶9(MMP9)及黏蛋白5AC(MUC5AC)的表达,采用Western blot检测各组大鼠泪腺中活化的半胱氨酸蛋白3(Cleaved-Caspase 3)、B淋巴细胞瘤2蛋白(Bcl2)、与Bcl2关联的X蛋白(Bax)、Bcl2相关的死亡促进蛋白(Bad)及基质金属蛋白酶9(MMP9)的表达。结果:与生理盐水组比较,4Z组、4E组和混合组大鼠眼表MUC5AC的表达增加,但MMP9的表达降低(P <0. 05);与4Z组比较,4E组大鼠眼表MUC5AC表达增加,但MMP9表达减少(P <0. 05或P <0. 01);与生理盐水组比较,4Z组、4E组和混合组大鼠泪腺中Cleaved-Caspase 3、Bax、Bad及MMP9表达降低,但Bcl2表达则增加(P <0. 05或P <0. 01);与4Z组相比,4E组大鼠泪腺中Cleaved-Caspase3、Bax及Bad的表达减少,但Bcl2表达增加(P <0. 05或P <0. 01)。结论:吉法酯增加干眼症大鼠眼表MUC5AC的表达,减少泪腺中Cleaved-Caspase3、Bax、Bad、Bcl及MMP-9的表达,且4E构型吉法酯和混合式吉法酯优于4Z构型吉法酯。
Objective: To investigate the protective mechanism of gefarnateon with different configurations on dry eye in rats. Methods: Forty Wistar female rats were chosen as the objects and eight were left as the normal group without any treatment. After being injected subcutaneously of scopolamine hydrobromide for 28 consecutive days in a high-circulation and low-humidity environment to prepare for a dry eye model,the 32 rats were divided into the normal saline group,the 4 Z gefarnate treatment group( 4 Z group),the 4 E gefardate treatment group( 4 E group),and the-mixed gifarfax treatment group( 4 Z ∶ 4 E = 30 ∶ 70,mixed group). The rats were executed by cervical dislocation on ice after 14 days of treatmen. Expression of MMP9 and MUC5 AC were detected by immunohistochemistry,and Western blot was used to detect the expression of activated Caspase-3,BCL2,Bax,Bad and MMP9 in the lacrimal gland. the expression of Cleaved-Caspase 3,Bax,Bad,Bcl2 and MMP9 in lacrimal gland by Western blot. Immunohistochemical staining was used to detect the expression of MMP9 and MUC5 AC in ocular surface. Results: Compared with the normal saline group,the expression of MUC5 AC and MMP9 of the 4 Z group,the 4 E group and the mixed group increased and reduced respectively( P < 0. 05 or P < 0. 01). Compared with the 4 Z group,the expression of MUC5 AC increased,but the expression of MMP9 decreased in the 4 E group( P < 0. 05 or P < 0. 01).Compared with the normal saline group,the expression of Cleaved-Caspase 3,Bax,Bad and MMP9 in the lacrimal gland of 4 Z group,4 E group and the mixed group decreased,but the expression of BCL2 increased( P < 0. 05 or P < 0. 01). Compared with the 4 Z group,the expression of Cleaved-Caspase3,Bax and Bad in the lacrimal gland of 4 E group decreased,but the expression of BCL2 increased( P <0. 05 or P < 0. 01). Conclusion: Gefarnate may increase the expression of MUC5 AC in ocular surface of rats with dry eye disease,and decrease the expression of Cleaved-Caspase3,Bax,Bad,Bcl and MMP-9 in the lacrimal gland. The gefarnate with 4 E configuration and the mixed configuration are better than that with the 4 Z configuration.

关键词(KeyWords): 吉法酯;炎症;细胞凋亡;干眼症;泪腺;机制
gefarnate;inflammation;apoptosis;dry eye;lacrimal gland;mechanism

Abstract:

Keywords:

基金项目(Foundation): 贵州省科学技术基金项目[黔科合JZ字(2015)2001];; 贵州省科技计划项目[黔科合支撑(2016)2848];; 贵州省教育厅青年科技人才成长项目[黔教合KY字(2017)155]

作者(Author): 糜玲;周波;李永;彭金刚;汤磊;黄家宇;
MI Ling;ZHOU Bo;LI Yong;PENG Jingang;TANG Lei;HUANG Jiayu;Department of Pharmaceutical Analysis,Guizhou Medical University;Guizhou Province Chemical Synthetic Drug Research and Development Engineering Technology Research Center;

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DOI: 10.19367/j.cnki.2096-8388.2020.06.006

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