川芎嗪对视网膜色素变性rd小鼠的干预作用及其抗凋亡作用机制研究

目的观察川芎嗪对视网膜色素变性以小鼠视网膜外核层细胞的病理、超微结构、凋亡和Bcl-2表达的影响，探讨川芎嗪对以小鼠的干预作用和干预机制。方法以新生小鼠84只随机分为实验组和对照组，每组42只。实验组小鼠从出生时腹腔注射盐酸川芎嗪80mg／kg，每天2次，至生后35天，对照组同时注射等量生理盐水。分别在用药后0、3、7、14、21、28、35天取眼球，立即经10％中性甲醛固定，常规病理切片。另取眼球经2．5％戊二醛溶液固定，电镜观察。用TUNEL方法检测视网膜外核层细胞的凋亡，用免疫组化方法测定Bcl-2在视网膜的表达。结果病理结果显示，经盐酸川芎嗪治疗后14、21、28、35天rd小鼠外核层细胞层数与未用药对照组比较，明显增加（P〈0．01）。电镜结果显示，川芎嗪可减缓以小鼠外核层细胞和视网膜光感受器细胞外段盘膜部位线粒体的病变，减少盘膜和外界膜的破坏。rd小鼠经盐酸川芎嗪药物治疗后第7、14、21、28、35天外核层细胞的凋亡率明显低于未用药对照组（P〈0．01）；第3、7、14、21、28、35天Bcl-2在视网膜外核层及光感受器细胞外段的表达明显增强（P〈0．05或0.01）。结论盐酸川芎嗪可延缓以小鼠视网膜外核层细胞的减少，其作用机制可能是通过上调视网膜外核层及光感受器细胞外段Bcl-2的表达延缓rd小鼠视网膜光感受器细胞的凋亡。

The interventional effect and anti-apoptosis mechanism of ligustrazine on the rd mice

OBJECTIVE To observe the effect of ligustrazine on anti - apoptosis of retina and Bcl - 2 expression of retina and explore the mechanisms of the intervention in rd mice.METHODS 84 of rd mice were randomly divided into 2 groups. Each group had 42 mice. Mice received twice a day ligustrazine by intraperitoneal injection from postnatal 1 to 35 days.At different time postnatal 1 day, 3 days, 7 days, 14 days, 21 days, 28 days, 35 days, the mice were sacrificed.The eyeballs were immediately enucleated and fixed in 10% formaldehyde in PBS.Routine pathologic process was performed. One eye in each group was fixed for the electron - microscopic examination. The photoreceptor cell apoptosis was detected by TUNEL technique and the inmmnohistochemistry method was used to detect Bcl - 2 expression in the retina. RESULTS Result from light microscopy showed that the layer number of retinal photoreceptor cell nuclei with ligustrazine therapy was increased at 14 days, 21 days, 28 days, 35 days in rd mice, compared with control group. Results from electron - microscope showed that ligustrazine could reduce lesions occurrence in the photoreceptor cells and the destruction in disc member, mitochondrion, and outer limiting membrane in the photoreceptor outer segment were decreased in rd mice by the comparison with control group. Compared with control group, cell numbers of the apoptotic photoreceptor cells were reduced and the Bcl - 2 expression in the matrix of retinal photoreceptors cell nuclei and photoreceptor outer segments were increased in experimental group in rd mice. CONCLUSIONS The ligustrazine uce retinal photoreceptor apoptosis by upregulating Bcl - 2 expression in the matrix of retinal photoreceptors cell nuclei or photoreceptor outer segments in rd mice.