枸杞多糖对蓝光损伤的视网膜色素上皮细胞的保护作用

Protective effects of lycium barbarum polysaccharide on damaged hRPE cells induced by blue light irradiation

摘要:

目的：研究不同浓度的枸杞多糖对蓝光诱导损伤的体外培养人视网膜色素上皮(human retinal pigment epithelium， hRPE)细胞的保护作用。

方法：通过蓝光诱导建立hRPE细胞光损伤模型，分别用不同浓度的枸杞多糖(分别为0.01，0.1，1mg/mL)对体外培养的 hRPE细胞进行干预，通过流式细胞仪检测各实验组的细胞线粒体活性氧和凋亡率。实验组分为正常对照组、光照损伤组以及不同浓度枸杞多糖(0.01，0.1，1mg/mL)干预组。

结果：线粒体活性氧检测：正常对照组荧光强度最小； 蓝光损伤组荧光强度最大，不同浓度枸杞多糖处理组荧光度与蓝光损伤组强度相比，荧光强度差异有统计学意义(P<0.05)。Annexin V-FITC/PI凋亡检测显示不同浓度枸杞多糖干预组凋亡细胞数量与蓝光损伤组凋亡细胞相比差异均有统计学意义(P<0.05)； 1mg/mL枸杞多糖干预组凋亡细胞数量与对照组凋亡数量相比差异无统计学意义(P>0.05)。

结论：枸杞多糖能抑制蓝光诱导损伤的hRPE细胞的凋亡，1mg/mL枸杞多糖抑制蓝光诱导的hRPE细胞凋亡的作用更强，其作用机制可能与抑制细胞线粒体产生活性氧有关。

Abstract:

AIM:To establish the model of age-related macular degeneration through blue light burning human retinal pigment epithelium(hRPE)cells in vitro and to investigate the possible mechanism of lycium barbarum polysaccharide(LBP)protecting the hRPE cells from blue light irradiation-induced damage.

METHODS: hRPE cells were cultured in Dulbecco's modified eagle's medium with high glucose and were pretreated with different concentrations(0.01mg/mL, 0.1mg/mL and 1mg/mL, respectively)of LBP solution, respectively. Further, hRPE cells were irradiated with different intensities(2000±500 LUX)of blue light(wave length 470-520nm)for 12h and the cellular morphology was observed by inverted phase contrast microscopy for every sample. Using flow cytometry, the alterations in the levels of reactive oxygen species, mitochondrial membrane potential and apoptosis were determined, respectively.

RESULTS: The level of reactive oxygen species in hRPE cells in blue-light treatment group was the highest, whereas the level in control group was the lowest. The fluorescent level of reactive oxygen species among different concentrations of LBP-treatment groups had specifically significant differences compared with that in blue light irradiation group(P<0.05). The amounts of apoptotic cells in different concentrations of LBP-treatment groups were specifically significant differences compared with that in blue light irradiation group(P<0.05); There were no apparent significant difference between 1mg/mL LBP-treatment group and normal control group(P>0.05).

CONCLUSION: LBP can efficiently inhibit apoptosis induced by blue light irradiation in hRPE cells and 1mg/mL of LBP has the strongest protective ability in blue light irradiation-induced damage in hRPE cells. The possible mechanism may attribute to the protective effect of LBP in inhibiting the overgeneration of reactive oxygen species and apoptosis in hRPE cells.