枸杞提取液对成人视网膜神经细胞活性的影响

Effect of extract of Lycium barbarum L. on adult human retinal nerve cells

目的研究枸杞提取液对视网膜神经细胞活性的影响。方法体外培养成人视网膜神经细胞，培养至24h和72h时，以MTT法检测细胞存活和生长情况；应用透射电镜观察细胞超微结构，测定细胞线粒体膜电位峰值，观察培养液内加入枸杞提取液后对成人视网膜神经细胞活性的影响。实验组中首次加药、再次加药及未加药对照组间细胞线粒体膜电位峰值比较，采用单因素方差分析；进一步两两比较采用LSD—t检验。结果细胞超微结构观察，显示加入枸杞提取液且培养两周的视网膜神经细胞外节段盘膜排列规整，内节段线粒体丰富，核染色质均匀。细胞培养至24h及72h时，MTT法检测，显示加入枸杞提取液的视网膜神经细胞增殖活跃，且呈剂量相关性，以加入枸杞提取液20g／L的保护作用最明显，细胞存活率分别为（223．23±12．13）％和（252．35±13．24）％。线粒体膜电位检测，显示实验组中首次加药、再次加药及未加药对照组的细胞线粒体膜电位峰值分别为36．35±9．44、47．99±11．10及3．83±1．30，组问差异有统计学意义（F=124．110，P〈0．05）；进一步两两比较，首次加入枸杞提取液后膜电位峰值较加药前增加了848％，差异有统计学意义（P=0．000）；再次加入枸杞提取液后膜电位峰值较加药前增加了1152％，差异有统计学意义（P=0．000）；表明枸杞提取液能够显著升高视网膜神经细胞的膜电位并使其兴奋性呈波动性变化。结论枸杞提取液对视网膜神经细胞有良好的保护作用，有望成为一种新的防治视网膜疾病的辅助治疗药物。

Objective To study possible protective effects of extract of Lycium barbarum L. on the euhured human retinal nerve cells. Methods Retinal nerve cells were co-cultured with the extract of Lycium barbarum L. and 24 hours and 72 hours later, retinal nerve ceils were respectively used to evaluate cell proliferation with MTT assays; to observe uhraeellular structural alternation with transmission electron microscopy （TEM） and to evaluate mitoehondrial membrane potentials （MMP） of cells with confoeal microscopy. The peaks of MMP between experiment group and control group were compared using one-way analysis of variance. Results Co-cultured retinal nerve cells with the extract were shown survival well under the TEM including photoreceptor segments remaining well, abundant mitoehondria in inner segments and well-distributed ehromatin in photoreceptor nucleus （F=124. 110, P 〈 0.05 ）. The addition of the extract promoted survival of adult retinal neurons significantly in concentration-dependent manner with the strongest effect in 20 g/L. Cell survival rate（24 h and 72 h） ; （223.23 ± 12. 13）% and （252. 35 ± 13.24）%. The peak of MMP increased 848% after the first adding of the extract （P =0. 000） and 1152% after the second adding of the extract （ P = 0. 000 ）. It showed that the extract could enhance the MMP significantly with undulatory property. Conclusions The extract of Lycium barbarum L. showed protective effects on cultured cells and could be used in treatment of some retinal diseases in future.