论著

海藻糖对实验性干眼病鼠结膜上皮细胞的影响

Effect of trehalose on conjunctival epithelial cells in a rat model of experimental dry eye

:141-149
 
目的:评价海藻糖滴眼液对摘除眶内眶外泪腺的实验性大鼠干眼的疗效。方法:SD大鼠30只,随机分成正常组、对照组、实验组,每组10只(20只眼)。正常组不给予处置,实验组和对照组通过摘除大鼠主泪腺制作干眼动物模型。对照组和实验组于造模完成术后1周后分别给予生理盐水、海藻糖滴眼液点双眼,每天4次,治疗共4周。分别于造模术前和造模术后1,2,3,5周进行基础泪液分泌试验(Shirmer I test),角膜上皮荧光染色评分检测;造模5周后将大鼠处死,采用免疫组织化学方法对结膜上皮细胞中Bcl-2和Bax凋亡基因蛋白表达进行检测,观察干眼动物模型结膜上皮细胞凋亡与眼表干眼病变及组织损伤的关系。结果:基础泪液分泌试验结果显示,对照组和实验组除术后第5周比较差异有统计学意义外(P<0.05),其余时间点差异无统计学意义(P>0.05),术后各时间点与正常组比较差异均有统计学意义(P<0.05)。造模术后第1周,对照组和实验组角膜荧光色染色评分明显高于正常组;术后第2周后,对照组和实验组评分差异无统计学意义(P>0.05),随着时间的延长,术后第3周和第5周,两组评分差异均有统计学意义(P<0.05),且实验组评分小于对照组。结膜上皮细胞免疫组织化学检测结果显示:正常组结膜上皮细胞Bax蛋白阳性表达的细胞数较对照组和实验组少,与对照组比较,差异有统计学意义(P<0.05),与实验组比较差异无统计学意义(P>0.05);正常组可见大量Bcl-2蛋白阳性表达,且多于实验组,差异无统计学意义(P>0.05),多于对照组,差异有统计学意义(P<0.05);对照组结膜上皮细胞中可见大量Bax蛋白阳性表达于细胞质,呈棕黄色颗粒,实验组中Bax蛋白阳性表达相对于对照组明显较少,两者差异有统计学意义(P<0.05);实验组结膜上皮细胞中可见大量Bcl-2蛋白阳性表达,对照组Bcl-2蛋白表达相对于实验组明显减少,差异有统计学意义(P<0.05)。结论:海藻糖滴眼液可以减少角膜表层上皮脱落,对结膜上皮细胞有一定的保护作用,可以拮抗结膜上皮细胞的凋亡,从而增强角结膜上皮屏障功能,改善由干燥引起的眼表上皮的损伤。海藻糖溶液可用于眼表上皮疾病的潜在临床应用,为治疗眼表疾病如干眼症提供治疗选择。
Objective: To observe the efficacy of trehalose eye drops on experimental dry eye rat with removal of inside and outside lacrimal glands. Methods: A total of 30 SD rats were randomly divided into a normal group, a control group, and an experimental group, with 10 rats for each group (20 eyes). The normal group was not treated, and the experimental group and the control group were made into a dry eye animal model by removing the main lacrimal gland of the rat. The control group and the experimental group were given with normal saline and trehalose eye drops once a week after the completion of modeling, 4 times a day for 4 weeks. Shirmer I test and fluorescence staining test were carded before and 1, 2, 3, and 5 weeks after modeling. After 5 weeks of modeling, the rats were sacrificed and the expression of Bcl-2 and Bax apoptosis protein in conjunctival epithelial cells was detected by immunohistochemistry in order to observe the relationship between apoptosis of conjunctival epithelial cells and dry eye lesions and tissue damage in dry eye animal models. Results: Schirme I test results showed that there was no significant difference between the control group and the experimental group (P>0.05) except for the 5th week after operation (P<0.05). There was a statistical difference between the two groups and the normal group at each time point after surgery (P<0.05). At the first week after modeling, the corneal fluorescence staining scores of the control group and the experimental group were significantly higher than those of the normal group. After the second week after surgery, there was no significant difference between the control group and the experimental group (P>0.05). The prolongation of time, the 3 week and the 5th week after surgery, the difference between the two groups was statistically significant (P<0.05), and the score of the experimental group was smaller than the control group. The immunohistochemical results of conjunctival epithelial cells showed that the number of Bax protein positive cells in the normal conjunctival epithelial cells was less than that in the control group and the experimental group. The difference between the normal group and the control group was statistically significant(P<0.05). There was no significant difference between the two groups (P>0.05). There was a large amount of Bcl- 2 protein in the normal group, which was more than the experimental group. The difference was not statistically significant (P>0.05), which was more than the control group. The difference was statistically significant (P<0.05); a large amount of Bax protein was positively expressed in the cytoplasm and brownish-yellow particles in the conjunctival epithelial cells of the control group. The positive expression of Bax protein in the experimental group was significantly less than that in the control group, and the difference was statistically significant (P<0.05). A large amount of Bcl-2 protein was positively expressed in the conjunctival epithelial cells of the experimental group, and the expression of Bcl-2 protein in the control group was significantly decreased compared with the experimental group (P<0.05).Conclusion: Trehalose eye drops can reduce epithelial detachment of the cornea and have a protective effect on conjunctival epithelial cells, which can antagonize the apoptosis of conjunctival epithelial cells,thereby enhancing the keratoconjunctival barrier function and improving the ocular surface epithelium caused by dryness. The potential clinical application of trehalose solution for ocular surface epithelial diseases and provide treatment options for ocular surface diseases, such as dry eye.
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  • 眼科学报

    主管:中华人民共和国教育部
    主办:中山大学
    承办:中山大学中山眼科中心
    主编:林浩添
    主管:中华人民共和国教育部
    主办:中山大学
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  • Eye Science

    主管:中华人民共和国教育部
    主办:中山大学
    承办:中山大学中山眼科中心
    主编:林浩添
    主管:中华人民共和国教育部
    主办:中山大学
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