摘要
目的:研究兔视网膜色素(RPE)上皮细胞光损伤以及枸杞子含药血清干预防治后细胞中参与凋亡的蛋白分子表达量的变化,探讨枸杞子含药血清对兔视网膜色素上皮细胞光损伤保护可能的信号传导途径,为临床应用枸杞子治疗年龄相关性黄斑变性等视网膜光损伤性疾病提供理论依据。方法:体外培养兔RPE细胞,用三基色冷光灯模拟自然光源,选取(2500±500)Lux作为基础光照强度,照射4h建立光损伤模型。制备含不同效量的枸杞子含药血清,光照前1h孵育体外培养的兔RPE细胞后进行光照实验,设置四个实验分组:A组:空白对照组;B组:光损伤模型组;C组:低剂量组;D组:高剂量组。通过透射电子显微镜进行兔RPE细胞超微结构的观察及拍片;采用蛋白免疫印迹(Western blot)法检测兔RPE细胞内细胞色素C(Cyt-C)、caspase-9及caspase-3蛋白的表达量。结果:与空白对照组相比,光损模型组细胞内的Cyt-C、Caspase-9、Caspase-3蛋白表达水平明显上调(P<0.05);枸杞子含药血清能有效的抑制细胞内Cyt-C、Caspase-9、Caspase-3蛋白的表达,与光损模型组比较,差异有统计学意义(均P<0.05),其中高浓度组可显著抑制细胞内Cyt-C、Caspase-9、Caspase-3蛋白的表达,与空白对照组比较,差异无统计学意义(P>0.05)。结论:枸杞子对RPE细胞具有一定的保护作用,这一保护作用可能是通过线粒体信号传导途径来实现的。
Objective ： To study the changes of rabbits retinal pigment epithelial cells participating in the expression of apoptotic protein molecule after photodamage and intervention of serum - containing medlar and discuss the molecular mechanism of Lycium barbarum polysaccharides on hRPE cells damage protection,providing a theoretical basis for treating age -related macular degen- eration and other retinal light damage diseases with clinical application of medlar. Methods ： Rabbit retinal pigment epithelial cells were cultured in vitro. The three color cold light lamp was used to simulate the natural light source, selecting （2500 ~ 500） Lux as the same light intensity and irradiating for 4 h to build the damage model. Produce different concentrations of medlar drug - containing serum. 1 h before lighting, the rabbits RPE cells were incubated in vitro. Set four experimental groups：Group A：the blank control group ; Group B ： the light injury model group ; Group C ： the low - dose group ; Group D ： the high - dose group. Through transmission electron microscope, we observed the cell uhrastructure of apoptosis after light damage and took X - ray photo of hRPE. Western blot （Western blot） method was used to detect the expressions of Cyt- C,caspase -9 and Caspase- 3 in hRPE cells. Results：Compared with the control group, the light injury model group＇s intracellular Cyt -C, Caspase -9 and Caspase -3 protein expression levels were significantl~ up - regulated （P 〈 0. 05 ）. Medlar - containing serum can effectively, inhibit the ex- pression of the intracellular Cyt -C, Caspase -9 and Caspase -3 protein. Compared with the light injury model group,the differ- ence was statistically significant （ all P 〈 0. 05 ） ,wherein the high concentrations significantly inhibited the expressions of intracel- lular Cyt - C, Caspase - 9 and Caspase - 3 protein. Compared with the blank control group, the difference was not statistically sig- nificant （ P 〉 0. 05 ）. Conclusion ： LBP through regulation of mitochondrial signaling pathway in the expression of these genes plays an effective protection on light -damage RPE cells.
出处
《辽宁中医杂志》
CAS
北大核心
2016年第11期2410-2414,共5页
Liaoning Journal of Traditional Chinese Medicine
基金
宁夏高等学校科研项目(NGY2013076)
国家自然科学基金项目(81360557)
关键词
光损伤
兔视网膜色素上皮细胞
枸杞子
含药血清
线粒体信号传导通路
photo damage
rabbit retinal pigment epithelial cells
Medlar
drug - containing serum
mitochondrial signal trans- duction pathway
