氨碘肽滴眼液对氧化应激性白内障的作用机制研究

Study on the mechanism of amiotide eye drop on the action of oxidative stress cataract

目的 探究氨碘肽滴眼液对白内障的治疗作用及机制。方法 使用过氧化氢对人晶状体上皮细胞(SRA01/04)进行处理,建立不同损伤程度氧化应激细胞模型。将细胞分为对照组(常规培养)、低浓度模型组(400μmol·L^(-1)过氧化氢)、低浓度模型+氨碘肽高浓度实验组(400μmol·L^(-1)过氧化氢+稀释20倍氨碘肽滴眼液)、低浓度模型+氨碘肽低浓度实验组(400μmol·L^(-1)过氧化氢+稀释50倍氨碘肽滴眼液)、高浓度模型组(800μmol·L^(-1)过氧化氢)、高浓度模型+氨碘肽高浓度实验组(800μmol·L^(-1)过氧化氢+稀释20倍氨碘肽滴眼液)、高浓度模型+氨碘肽低浓度实验组(800μmol·L^(-1)过氧化氢+稀释50倍氨碘肽滴眼液)。检测细胞形态并对细胞存活率、细胞内活性氧(ROS)及细胞中增殖细胞核抗原(PCNA)蛋白表达的变化情况进行检测。结果 对照组、低浓度模型+氨碘肽低浓度实验组、高浓度模型+氨碘肽低浓度实验组、低浓度模型组、高浓度模型组的SRA01/04细胞ROS水平分别为318.00±7.00,374.67±14.98,810.00±25.71,608.67±11.59和932.33±30.55;这5组的细胞增殖活力分别为(100.00±2.29)%,(97.24±2.86)%,(63.12±3.91)%,(87.05±2.96)%和(57.28±3.62)%;这5组的PCNA蛋白相对表达水平分别为1.06±0.03,0.91±0.11,0.66±0.06,0.80±0.07和0.49±0.05。上述指标,低浓度模型、高浓度模型组与对照组比较,差异均有统计学意义(均P<0.05);低浓度模型+氨碘肽低浓度实验组与低浓度模型组比较,差异均有统计学意义(均P<0.05);高浓度模型+氨碘肽低浓度实验组与高浓度模型组比较,除细胞增殖活力指标,其余数据差异均有统计学意义(均P<0.05)。结论 氨碘肽滴眼液具有较好的生物相容性,能有效降低细胞内源性活性氧,提高细胞的存活率,增加细胞内PCNA蛋白的表达,进而增强细胞增殖活力。

Objective To explore the therapeutic effect and mechanism of amiotide eye drops on cataract.Methods To establish oxidative stress models with different degrees of cell damage in vitro,human lens epithelial cells (SRA01/04) were treated with hydrogen peroxide at different concentrations.The cells were divided into several groups:control group (conventional culture),model-L group(400μmol·L^(-1)H2O2),model-L+experimental-H group(400μmol·L^(-1)H2O2+amiotide eye drop with 20 dilution),model-L+experimental-L group(400μmol·L^(-1)H2O2+amiotide eye drop with 50 dilution),modelH group(800μmol·L^(-1)H2O2),model-H+experimental-H group(800μmol·L^(-1)H2O2+amiotide eye drop with 20 dilution),and model-H+experimental-L group(800μmol·L^(-1)H2O2+amiotide eye drop with 50 dilution).Cell morphology,cell viability and the variation of the value of intracellular reactive oxygen species (ROS) and proliferating cell nuclear antigen (PCNA) protein expression were respectively measured.Results The strength of ROS in control group,model-L+experimental-L group,modelH+experimental-L group,model-L group,and model-H group were 318.00±7.00,374.67±14.98,810.00±25.71,608.67±11.59 and 932.33±30.55,respectively.The cell proliferation rate in the above 5 groups were (100.00±2.29)%,(97.24±2.86)%,(63.12±3.91)%,(87.05±2.96)%and (57.28±3.62)%,respectively.The protein expression level of PCNA in the above 5 groups were 1.06±0.03,0.91±0.11,0.66±0.06,0.80±0.07 and 0.49±0.05,respectively.Compared between model-L,model-H groups and normal group,the difference of the above indexes were significant (all P<0.05).Compared between model-L+experimental-L group and model-L group,the differences of the above indexes were significant (P<0.05).Comparison between model-H+experimental-L group and model-H group,except for the index of cell proliferation activity,other indexes were statistically significant (P<0.05).Conclusion Amiotide eye drops have good biocompatibility.It could reduce endogenous ROS,inducing the cell survival rate improvement,and the expression of PCNA protein increasing,which further showed a better cell proliferation vitality.