枸杞多糖对体外培养的视网膜神经节细胞氧化应激损伤的保护作用

Effects of lycium barbarum polysaccharides therapy on oxidative stress-induced apoptosis of cultured retinal ganglion cells

目的观察枸杞多糖对体外培养的视网膜神经节细胞氧化应激损伤的保护作用。方法实验分为正常对照组、H_2O_2组、H_2O_2+LBP(40 mg/L)组。正常对照组:DMEM完全培养基常规培养;H_2O_2组:含体积分数10%FBS的DMEM高糖培养基培养,同时加入200μmol/L H_2O_2溶液造成氧化应激损伤模型;H_2O_2+LBP(40 mg/L)组:用含10%FBS和200μmol/L H_2O_2的DMEM培养基培养,同时加入不同浓度的枸杞多糖(10 mg/L,20 mg/L,40 mg/L)进行干预,最终选用40 mg/L枸杞多糖进行保护。使用四甲基偶氮唑盐(MTT)法和Annexin V-FITC法检测细胞存活率及凋亡率。结果 MTT结果显示,H_2O_2能够降低RGC-5细胞活性,并呈浓度依赖性,200μmol/L H_2O_2作用24 h后RGC-5细胞活性为正常对照组的50%,继续加入不同浓度LBP(10 mg/L,20 mg/L,40 mg/L)干预后细胞存活率分别为52.5%、62.8%和68.6%,与H_2O_2组相比较细胞存活率明显升高,差异有统计学意义(P<0.05)。Annexin V-FITC检测结果显示,正常对照组、H_2O_2组、H_2O_2+LBP(40 mg/L)组细胞凋亡率,分别为4.90%、32.35%、24.21%,各组间差异有统计学意义(P<0.05)。结论枸杞多糖对RGC-5氧化应激损伤有保护作用。

Objective To investigate the neuroproteetive effects of lycium barbarum polysaccharides （LBP） against oxidative stress - induced injury in cultured retinal ganglion cells. Methods Rat RGC line RGC -5 were cuItured in vitro, and were randomly di- vided into the normal group,H202 group and H2O2 ＋ LBP （40rag/L） group. Then apoptosis models were cultured a series concentration of LBP （ 10 mg/L,20 mg/L and 40 mg/L）. Cell viability of the RGC - 5 cell line was measured with MTT assay ,flow cytometry was used to detect the apoptosis process. Results The results of MTT eolorimetry showed that H2 O2 may induce cell death of RGC -5 cells in a concentration - dependent manner, exposure to 200 p, moVL H2O2 clearly induced cell death in 50% of the cells within 24 hours, the sur- vival ratio after exposure to 200μmol/L H2O2 was lower than that in control cell. The survival ratio in LBP group （ 10 mg/L,20 mg/L,40 mg/L） were 52.5% ,62.8% and 68.6%, respectively. There was statistical difference between 10 mg/L,20 mg/Land 40 mg/L group （P 〈 0.05）. Flow cytometry showed apoptosis ratio of RGC- 5 in control group, H2O2 group and H2O2 ＋ LBP group were respectively 4.90% ,32.35% and 24.21%, （P 〈 0.05）. Conclusion The study demonstrates that LBP can prevent RGC- 5 from apoptosis.