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[通过细胞荧光光度法测定核DNA研究大鼠晶状体上皮细胞动力学]

[Cell kinetics of rat lens epithelium by cytofluorometric nuclear DNA determination].

作者信息

Takayanagi K, Kubo E, Tsuzuki S, Kobayashi T, Mori K, Takahashi Y, Akagi Y

机构信息

Department of Ophthalmology, Fukui Medical School, Japan.

出版信息

Nippon Ganka Gakkai Zasshi. 1995 Oct;99(10):1127-32.

PMID:8533634
Abstract

The cell kinetics of rat lens epithelium was assessed by measuring the changes in the nuclear DNA contents during sugar cataract formation. Six-and-12-week-old Sprague-Dawley male rats were used and divided into the following groups: fed on normal chow, fed on 25% galactose diet, and fed on normal chow after 5 days on the 25% galactose diet. Every second day following the beginning of each chow feeding, lenses were extracted, and lens capsules with epithelial cells were obtained. After a few day's fixation in 4% paraformaldehyde dissolved in 0.1M phosphate buffer (pH 7.4), DAPI (4'6-diamidino-2-phenylindole)-stained lens epithelial cells were measured by fluorescence cytophotometry. The epithelia in the normal chow-fed rats contained many 2C and a few 4C nuclei. In the 25% galactose-fed rats, 4C nuclei increased gradually in number until the fifth day, and then decreases slowly day by day. Abnormal polyploid nuclei (8C) were observed in the 25% galactose-fed rats. 4C nuclei decreased rapidly after the diet reversal. These results indicate that galactose feeding caused higher DNA synthesis of rat lens epithelial cells and a higher possibility of abnormal cell division.

摘要

通过测量糖性白内障形成过程中核DNA含量的变化来评估大鼠晶状体上皮细胞的动力学。选用6周龄和12周龄的Sprague-Dawley雄性大鼠,分为以下几组:喂普通饲料组、喂25%半乳糖饲料组以及在喂25%半乳糖饲料5天后再喂普通饲料组。在每种饲料喂养开始后的第二天,取出晶状体,获取带有上皮细胞的晶状体囊膜。在溶于0.1M磷酸盐缓冲液(pH 7.4)的4%多聚甲醛中固定几天后,通过荧光细胞光度法测量用4',6-二脒基-2-苯基吲哚(DAPI)染色的晶状体上皮细胞。喂普通饲料的大鼠上皮细胞含有许多2C核和少数4C核。在喂25%半乳糖的大鼠中,4C核数量逐渐增加直至第5天,然后逐日缓慢减少。在喂25%半乳糖的大鼠中观察到异常多倍体核(8C)。饮食逆转后4C核迅速减少。这些结果表明,喂半乳糖会导致大鼠晶状体上皮细胞的DNA合成增加以及细胞异常分裂的可能性增加。

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