| 雷帕霉素在兔眼滤过性手术后的抗瘢痕作用 |
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| 引用本文： | 邰雪,申颖,赵海霞,王召格,关文英,康欣,郭文奇. 雷帕霉素在兔眼滤过性手术后的抗瘢痕作用[J]. 南方医科大学学报, 2020, 40(9): 1346-1352. DOI: 10.12122/j.issn.1673-4254.2020.09.19 |
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| 作者姓名： | 邰雪 申颖 赵海霞 王召格 关文英 康欣 郭文奇 |
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| 摘 要： | 目的 研究雷帕霉素在兔眼滤过性手术后抗瘢痕形成的作用及其产生作用可能的原因。方法 取健康成年的青紫兰兔96只,取每只兔左眼行小梁切除术,术后随机分为实验组及对照组,24只/组。术后将配制好的不同浓度雷帕霉素滴眼剂（1%、3%、5%）用于实验组,将药用蓖麻油滴眼剂用于实验对照组。4次/d。术后分别于第7、14、21、28天依照Krofeld分型法比较滤过泡的形态和功能。在术后第1、2、3、4周分别随机处死6只青紫蓝兔后,采用免疫组化法检测兔眼行小梁切除术区域的组织中增殖细胞核抗原（PCNA）和α-平滑肌肌动蛋白（α-SMA）因子局部表达情况,观察结果。体外原代培养兔眼结膜下成纤维细胞（RTFs）,采用流式细胞技术检测经不同浓度雷帕霉素溶液（0、0.06、0.25、1、4 mg/L）作用后细胞的凋亡情况。结果 在术后第1、2、3周可见各实验组功能性滤过泡形成率比对照组高（P<0.05）,且功能性滤过泡的形成率呈雷帕霉素药物浓度依赖性增长;免疫组化法可见实验组PCNA标记阳性成纤维细胞数表达量比相应的对照组高,各实验组数目比相应的对照组少（P<0.05）,且α-SMA阳性成纤维细胞的表达程度随时间延长呈下降趋势;体外培养RTFs后,用不同浓度雷帕霉素溶液（0、0.06、0.25、1、4 mg/L）作用于RTFs,流式细胞技术检测可见RTFs在各药物浓度下均出现凋亡,当浓度为0.25 mg/L时凋亡出现明显的增加（P<0.05）,并且RTFs的凋亡呈浓度依赖性。结论 雷帕霉素可以抑制兔青光眼术后滤过道组织过度增生,可较多的保留功能性滤过泡,并延长其寿命;雷帕霉素可以有效的诱导RTFs凋亡的发生,从而产生抑制青光眼术后滤过区成纤维细胞过度增殖的作用。
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| 关 键 词： | 雷帕霉素 青光眼滤过性手术 增殖细胞核抗原 α-平滑肌肌动蛋白 信号转导通路 |
Anti-scarring effect of rapamycin following filtering surgery in rabbit eyes |
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| Abstract: | Objective To study the effect of rapamycin on scar formation in rabbit eyes following filtering operation and explorethe possible mechanism. Methods Ninety-six healthy adult rabbits were subjected to trabeculectomy of the left eye andsubsequently randomly divided into 4 groups (n=24) for treatment with castor oil (control) or rapamycin (1%, 3%, or 5%) eyedrops of the operated eyes 4 times a day. The morphology and function of the filtering blebs of the rabbits were compared at 7,14, 21 and 28 days after the operation; at each of the time points, 6 rabbits from each group were euthanized for detection ofexpressions of proliferating cell nuclear antigen (PCNA) and α-smooth muscle actin (α-SMA) in the tissues in the surgical areausing immunohistochemistry. Cultured rabbit subconjunctival fibroblasts (RTFSs) were treated with different concentrations ofrapamycin (0.06, 0.25, 1, and 4 mg/L) and the cell apoptosis was detected using flow cytometry. Result In the first, second andthird weeks after the operation, the rate of functional follicle formation was significantly higher in the 3 rapamycin groupsthan in the control group (P<0.05), and the number of α- SMA-positive fibroblasts decreased over time in the 3 rapamycingroups. In cultured RTFSs, treatment with rapamycin at different concentrations resulted in increased apoptosis of the cells,and rapamycin above 0.25 mg/L significantly increased the cell apoptosis in a dose-dependent manner. Conclusion Rapamycincan inhibit hyperplasia of the filtering passage tissue, helps to preserve the functional filtering blebs and prolong their lifespan, and induces apoptosis of RTFS. |
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| Keywords: | rapamycin glaucoma filtering operation proliferating cell nuclear antigen α-smooth muscle actin signaltransduction |
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