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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 protein expression in apoptosis of retinal pigment epithelium (RPE) epithelial cells and the apoptosis of RPE-containing serum after the intervention of medicated serum containing medlar, and to explore the protective effect of medlar containing serum on the damage of retinal pigment epithelial cells in rabbits Possible signal transduction pathways for the clinical application of medlar treatment of age-related macular degeneration retinal photodamage damage and other diseases provide a theoretical basis. Methods: Rabbit RPE cells were cultured in vitro. The natural light source was simulated by using three-color cold light. The light intensity was selected as (2500 ± 500) Lux and the light damage model was established by irradiation for 4 hours. Preparation of drug-containing serum containing different effective concentrations of medlar 1h incubation before in vitro cultured rabbit RPE cells were light experiments, set up four experimental groups: A group: blank control group; B group: light injury model group; C group : Low dose group; D group: high dose group. The ultrastructure of rabbit RPE cells was observed by transmission electron microscopy and photographed. The expression of cytochrome C (Cyt-C), caspase-9 and caspase-3 in rabbit RPE cells were detected by Western blot . Results: Compared with the blank control group, the expression of Cyt-C, Caspase-9 and Caspase-3 in the model group were significantly increased (P <0.05); Medlar serum containing Lycium barbarum could effectively inhibit the intracellular Cyt The expression of Caspase-9, Caspase-9, Caspase-3, Caspase-3 and Caspase-3 were significantly different from those of the model group (all P < Caspase-3 protein expression, compared with the blank control group, the difference was not statistically significant (P> 0.05). CONCLUSION: Fructus Lycii has a protective effect on RPE cells, and this protective effect may be achieved through the mitochondrial signal transduction pathway.