川芎嗪对慢性高眼压大鼠视网膜神经节细胞的保护作用研究

Study on the protective effect of ligustrazine on retinal ganglion cells in rats with chronic ocular hypertension

目的探讨川芎嗪对慢性高眼压大鼠视网膜神经节细胞的保护作用。方法将36只健康雄性SD大鼠随机分为正常组、高眼压组和川芎嗪组,每组12只。正常组正常饲养,高眼压组和川芎嗪组应用巩膜静脉烧灼法建模。在大鼠高眼压持续1周后,川芎嗪组给予川芎嗪腹腔注射,正常组和高眼压组给予等量生理盐水腹腔注射,1次/d,连续14 d。实验结束后取大鼠眼球,HE染色观察视网膜形态,TUNEL染色检测视网膜神经节细胞凋亡情况,免疫组化法检测视网膜神经节细胞层中bcl-2蛋白表达情况。结果建模后1周、2周、3周、4周,高眼压组和川芎嗪组眼压值均明显高于正常组(P均<0.05),高眼压组和川芎嗪组眼压值比较差异均无统计学意义(P均>0.05)。HE染色显示,正常组视网膜各层次呈现规整排列,视网膜神经节细胞单层排列;高眼压组视网膜神经节细胞减少,视网膜萎缩;川芎嗪组视网膜结构层次清晰,视网膜神经节细胞表现与正常组类似,视网膜损伤显著减轻。正常组大鼠视网膜无TUNEL阳性细胞,川芎嗪组TUNEL阳性细胞数显著少于高眼压组(P<0.05)。免疫组化检测显示,正常组大鼠视网膜神经节细胞层中可见bcl-2弱阳性表达;高眼压组bcl-2表达有所增强,光密度值明显高于正常组(P<0.05);川芎嗪组bcl-2表达明显增强,光密度值明显高于高眼压组(P<0.05)。结论川芎嗪能够通过促进bcl-2的表达而抑制视网膜神经节细胞的凋亡并起到保护作用。

Objective It is to investigate the protective effect of ligustrazine on retinal ganglion cells （RGCs） in rats with chronic ocular hypertension. Methods 36 healthy male rats were randomly divided into group A （ normal group） , group B （high intraocular pressure group） and group C （ligustrazine treatment group）, 12 rats in each group. The rats were fed nor- mally in group A, and were treated with scleral vein cauterization to make high intraocular pressure model of rats in Group B and group C. After 1 week of high intraocular pressure persistent state in rats, the animals were given intraperitoneal injection with ligustrazine in Group C,and with normal saline in group A and group B, once a day, continuous treatment for 14 days. At the end of the experiment, the eye of rats were taken and the morphology of retina was observed by HE staining, and the apop- tosis of retinal ganglion cells was detected by TUNEL staining, the expression of bcl-2 protein in retinal ganglion cell layer was detected by immunohistochemical method. Results After 1-, 2-, 3-and 4-week of modeling, the intraocular pressure of group B and group C were significantly higher than those of group A（ all P 〈 0.05 ） , there was no significant difference in intraocular pressure between group B and group C（ all P 〉 O. 05 ）. HE staining showed that the arrangement of the retina was regular, and the retinal ganglion cells were arranged in a single layer in group A; the retinal ganglion cell count was reduced, and retina was atrophied in group B ; the retinal structure was clear, and the retinal ganglion cells was similar to that of group A, the reti- nal damage was significantly reduced in group C. There was no TUNEL positive cells in rats of Group A, the TUNEL positive cells count in group C was significantly decreased than that of Group B （ P 〈 0.05 ）. The immunohistochemical detection showed, the expression of bcl-2 was weakly positive in the retinal ganglion ceils of rats in group A ; the expression of bcl-2 was increased in group B, the optical density was significantly higher than group A（ P 〈 0.05 ） ; the expression of bcl-2 was signifi-cantly enhanced in group C, the optical density was significantly higher than group B （P 〈 0. 05）. Conclusion ligustrazine can inhibit the apoptosis of retinal ganglion cells and play a protective role by regulating the expression of bcl-2.