Talebizadeh Nooshin, Yu Zhaohua, Kronschläger Martin, Söderberg Per
Gullstrand Lab, Ophthalmology, Department of Neuroscience, University of Uppsala, Uppsala, Sweden.
Acta Ophthalmol. 2014 Dec;92(8):769-73. doi: 10.1111/aos.12407. Epub 2014 Apr 3.
To determine the time evolution of active caspase-3 protein expression in albino rat lens after in vivo exposure to low-dose UVR-300 nm, as detected by immunofluorescence.
Forty Sprague-Dawley rats were unilaterally exposed in vivo to 1 kJ/m(2) UVR-300 nm for 15 min. At 0.5, 8, 16 and 24 hr after the UVR exposure, the exposed and contralateral nonexposed lenses were removed and processed for immunohistochemistry. Three mid-sagittal sections from each lens were stained. The cells labelled for active caspase-3 in each section of both the exposed and nonexposed lenses were counted and recorded three times. The difference of the proportion of labelling between the exposed and contralateral nonexposed lenses within each animal was calculated. The differences of active caspase-3 labelling at four different time-points after exposure were used to determine the time evolution of active caspase-3 expression.
Caspase-3 expression was higher in the exposed than in contralateral nonexposed lenses. The mean difference between the exposed and contralateral nonexposed lenses, including all lenses from all time intervals, was 0.12 ± 0.01 (= CI 95%). The mean differences between the exposed and contralateral nonexposed lenses were 0.11 ± 0.02, 0.13 ± 0.02, 0.14 ± 0.01 and 0.09 ± 0.03 (= CI 95%) for the 0.5-, 8-, 16- and 24-hr time groups, respectively. The orthogonal comparison showed no difference in the expression of active caspase-3 between the 0.5- and the 24-hr groups (Test statistic 1.50, F1,36 = 4.11, p < 0.05) or between the 8- and the 16-hr groups (test statistic 0.05, F1,36 = 4.11, p < 0.05). There was a difference when comparing the 0.5- and 24-hr groups to the 8- and 16-hr groups (test statistic 7.01, F1,36 = 4.11, p < 0.05).
The expression of active caspase-3 in the lens epithelium increases after UVR exposure. There is a peak of expression approximately 16 hr after the exposure.
通过免疫荧光检测，确定白化大鼠晶状体在体内暴露于低剂量300nm紫外线辐射（UVR）后，活性半胱天冬酶-3蛋白表达随时间的变化情况。
40只Sprague-Dawley大鼠单侧体内暴露于1kJ/m(2)的300nm紫外线辐射15分钟。在紫外线辐射后0.5、8、16和24小时，取出暴露侧和对侧未暴露的晶状体，进行免疫组织化学处理。每个晶状体取三个中矢状切片进行染色。对暴露侧和未暴露侧晶状体各切片中标记有活性半胱天冬酶-3的细胞进行计数并记录三次。计算每只动物暴露侧和对侧未暴露侧晶状体标记比例的差异。利用暴露后四个不同时间点活性半胱天冬酶-3标记的差异来确定活性半胱天冬酶-3表达随时间的变化情况。
暴露侧晶状体中半胱天冬酶-3的表达高于对侧未暴露的晶状体。暴露侧和对侧未暴露侧晶状体的平均差异，包括所有时间间隔的所有晶状体，为0.12±0.01（=95%置信区间）。0.5小时、8小时、16小时和24小时时间组中，暴露侧和对侧未暴露侧晶状体的平均差异分别为0.11±0.02、0.13±0.02、0.14±0.01和0.09±0.03（=95%置信区间）。正交比较显示，0.5小时组和24小时组之间活性半胱天冬酶-3的表达无差异（检验统计量1.50，F1,36 = 4.11，p < 0.05），8小时组和16小时组之间也无差异（检验统计量0.05，F1,36 = 4.11，p < 0.05）。将0.5小时组和24小时组与8小时组和16小时组进行比较时存在差异（检验统计量7.01，F1,36 = 4.11，p < 0.05）。
紫外线辐射后晶状体上皮中活性半胱天冬酶-3的表达增加。暴露后约16小时表达出现峰值。
