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金属硫蛋白与大鼠放射性白内障的关系

发布时间:2018-04-28 03:06

  本文选题:X射线 + 放射性白内障 ; 参考:《苏州大学》2012年硕士论文


【摘要】:目的: 利用动物实验,通过一次性大剂量(25Gy) X射线照射S-D大鼠双眼建立放射性白内障模型,观察照射后一周、二周、四周时大鼠晶状体情况,同时检测其晶状体金属硫蛋白(Metallothionein,MT)的表达水平,探讨金属硫蛋白与放射性白内障形成之间的关系。 方法: 照射前选取8只大鼠作为正常对照组。实验照射组大鼠24只,一次性照射X射线(双眼同时照射)25Gy。所有大鼠分别于照射后一周、二周、四周时,腹腔注射3.6%水合氯醛麻醉后,用托吡卡胺滴眼液充分散瞳,裂隙灯显微镜观察并记录晶状体混浊情况;然后在上述每个时间点的实验照射组依次随机选取8只大鼠处死取其晶状体。分别运用逆转录聚合酶链反应(Reverse Transcription-Polymerase Chain Reaction,RT-PCR)和免疫印迹法(Western blotting)检测MT-Ⅰ、MT-ⅡmRNA和金属硫蛋白(MT)在上述时间点大鼠晶状体内的表达情况。 应用SPSS17.0统计软件对结果进行统计分析,采用Independent Samples T-Test法和One-Way-ANOVA法分别对计量资料进行独立样本T检验和单因素方差分析,并使用LSD法进行组间比较。 结果: 1.形态学:一次性大剂量(25Gy)X射线照射大鼠双眼,于照射后一周、二周时散瞳观察大鼠晶状体均未见混浊,而照射后四周观察时可见晶状体后囊和后皮质开始出现细小、散在白点状混浊。 2.RT-PCR结果显示:在mRNA水平MT-Ⅰ、MT-Ⅱ在大鼠晶状体上均有表达,实验组和对照组的晶状体进行RT-PCR均可获得MT-Ⅰ、MT-Ⅱ的阳性目的基因。实验组晶状体的MT-Ⅰ、MT-ⅡmRNA的表达均较对照组均有不同程度的增加,差异具有统计学意义(P0.05)。其变化是随着照射后时间的延长,上述时间点MT-Ⅰ、MT-ⅡmRNA的表达呈先上升后下降的趋势,各时间点的变化有差异,差异具有统计学意义(P0.05)。 3. Western blotting结果显示:实验照射组金属硫蛋白含量相对于实验对照组增加,差异有统计学意义(P<0.05);实验照射组金属硫蛋白含量的变化随照射时间的延长呈先增加后降低的趋势,并且上述时间点的变化有差异,其差异有显著统计学意义(P<0.05)。 结论: 一次性大剂量X射线(25Gy)照射大鼠双眼可在短时间内即可引起晶体混浊;同时它可提高晶状体MT-Ⅰ、MT-ⅡmRNA及MT蛋白的表达水平;当MT表达下降时晶体出现混浊,说明MT可能参与放射性白内障的形成。
[Abstract]:Objective: In animal experiment, the model of radiation-induced cataract was established in the eyes of S-D rats by X-ray irradiation with a large dose of 25 Gy at one time. The lens of the rats was observed one week, two weeks and four weeks after irradiation. At the same time, the expression level of metallothionein MTA was detected to explore the relationship between metallothionein and radiation cataract formation. Methods: Eight rats were selected as normal control group before irradiation. Twenty-four rats in the experimental group were exposed to X ray at one time (25 Gy for both eyes at the same time). All the rats were injected intraperitoneally with 3.6% chloral hydrate anesthetized one week, two weeks and four weeks after irradiation. The lens opacity was observed and recorded by slit lamp microscope. Then 8 rats were randomly selected at each time point in the experimental group to get their lens. Reverse transcriptase polymerase chain reaction (reverse Transcription-Polymerase Chain reaction) and Western blotting were used to detect the expression of MT- 鈪,

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