17β-雌二醇对人晶状体上皮细胞的保护作用与细胞焦亡的相关性研究
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泸州市科技计划项目\〖No.2015LZCYD-S05(4/12)\〗


Study on the relationship between the protective effect of 17β-estradiol on human lens epithelial cells and cell pyroptosis
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Luzhou Science and Technology Project \〖No.2015LZCYD-S05(4/12)\〗

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    摘要:

    目的:探索17β-雌二醇(17β-estradiol,E2)对人晶状体上皮细胞的保护作用与细胞焦亡的相关性及其机制。

    方法:体外培养人晶状体上皮细胞,设立空白对照组、H2O2处理组(含100μmol/L H2O2培养基培养12h)、雌二醇组(分别于含1×10-3、1×10-2、1×10-1μmol/L雌二醇培养基中培养24h后,再加入含100μmol/L H2O2的培养基处理12h)。扫描电子显微镜观察细胞超微结构; 免疫荧光染色法检测Gasdermin D(GSDMD)分布及荧光强度; CCK-8法检测细胞活力; TUNEL法检测细胞焦亡; 免疫印迹实验(Westen-blot)检测半胱氨酸天冬氨酸特异性蛋白水解酶-1(Caspase-1)、GSDMD、NOD样受体蛋白(NLRP3)表达水平; 酶联免疫吸附试验(ELISA)检测白介素-1β(IL-1β)表达。

    结果:与空白对照组相比,H2O2处理组的细胞活力下降,细胞焦亡率升高,Caspase-1、GSDMD、NLRP3蛋白的表达上调,IL-1β分泌增多。而与H2O2处理组相比较,雌二醇组细胞焦亡率下降,Caspase-1、GSDMD、NLRP3蛋白的表达下调,IL-1β分泌量减少,并随雌激素浓度的递增,而呈递减趋势。

    结论:17β-雌二醇对人晶状体上皮细胞的保护作用具有浓度依赖性,其保护机制与抑制人晶状体上皮细胞焦亡过程相关,并有经典焦亡途径参与。

    Abstract:

    AIM: To explore the relationship between the protective effect of 17β-estradiol(E2)on human lens epithelial cells and pyroptosis.

    METHODS: Human lens epithelial cells were cultured in vitro and divided into blank control group, H2O2 treatment group, and 17β-estradiol+H2O2 treatment group. Scanning electron microscope to observe the cytological morphology; immunofluorescence technique to detect Gasdermin D(GSDMD)distribution and fluorescence intensity; CCK-8 to detect cell viability; TUNEL to detect cell pyroptosis; Western-blot to detect Cysteinylaspartate specific proteinase-1(Caspase-1), GSDMD, NOD-like receptor protein 3(NLRP3)protein expression level; ELISA to detect interleukin-1β(IL-1β)expression.

    RESULTS: Compared with the control group, the cell viability of the H2O2 treatment group was significantly decreased, the expression of Caspase-1, GSDMD, and NLRP3 protein were significantly up-regulated, and the secretion of IL-1β was significantly increased. Compared with the H2O2 treatment group, the expression of Caspase-1, GSDMD, and NLRP3 protein in the 17β-estradiol+H2O2 treatment group were down-regulated, and the secretion of IL-1β decreased, and it showed a decreasing trend with the increase of estrogen concentration.

    CONCLUSION: 17β-estradiol has a protective effect on human lens epithelial cells, and its protective mechanism is related to the inhibition of the pyroptosis process of human lens epithelial cells, and the classical pyroptosis pathway is involved.

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陈颖,康刚劲,王妍茜,等.17β-雌二醇对人晶状体上皮细胞的保护作用与细胞焦亡的相关性研究.国际眼科杂志, 2021,21(10):1680-1685.

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  • 收稿日期:2021-03-26
  • 最后修改日期:2021-08-26
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  • 在线发布日期: 2021-09-16
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