灯盏花中单体成分对体外培养大鼠视网膜神经细胞的影响

Effect of caffeic acid,seopoletin and scutellarin on rat retinal neurons in vitro

目的:考察咖啡酸、东莨菪内酯、野黄芩苷对体外培养视网膜神经细胞存活的影响,探讨灯盏花中具视神经保护作用的有效成分。方法:采用胰酶消化法将18只出生2-3d的乳鼠视网膜制成细胞悬液,接种于经多聚鸟氨酸(HA)和层粘连蛋白(LN)包被的96孔板中。于培养3d后,分别加入PBS液及不同含量的咖啡酸、东莨菪内酯、野黄芩苷溶液继续培养2d,采用MTT比色法测量其存活细胞的吸收值,同时对部分细胞行Nissel体染色检查。结果:Nissel体染色检查结果表明,培养5d的存活细胞大部分均为神经细胞。与空白对照组比较,咖啡酸含量在3.9-1000μg·mL-1、东莨菪内酯、野黄芩苷含量在250-1000μg·mL-1时,其吸收值均明显增加(P<0.05,P<0.01),且有一定的量效关系。结论:咖啡酸、东莨菪内酯、野黄芩苷三者均能促进体外培养的视网膜神经细胞存活,且咖啡酸活性最强。

Objective: To observe the effect of caffeic acid, seopoletin and scutellarin on rat retinal neurons in vitro and explore neu-roprotection in glaucoma of Erigeron breviscapus. Method: The retinal of 18 post-natal 2-3 days Sprague-Dawley rats were dissociated into cell suspension with trypsin digestion. The cell suspension was implated in 96-well culture plates covered with hyaluronic acid and laminin in each well. After culturing for 3 days, caffeic acid, seopoletin and scutellarin were added to the cultures, continue to culture 2 days. Then, the A of living cells in each well was tested by M'lT colorimetric microassay. Some of the 5-day culture cells were identified by Nissel technique. Result: Most of the living cells were retinal neurons by Nissel identification. The number of living cells increased significantly in high concentrations of caffeic acid, seopoletin and scutellarin compared with control group(P < 0.05, P < 0.01) . Conclusion: caffeic acid, seopoletin and scutellarin can all promote retinal neurons to live in vitro, with caffeic acid being most effective.