[1]崔铮,靖鹏举,朱冬梅,等.地塞米松对脂多糖诱导人视网膜色素上皮细胞炎症损伤及NLRP3/Caspase-1通路的影响[J].眼科新进展,2020,40(9):826-830.[doi:10.13389/j.cnki.rao.2020.0188]
　CUI Zheng,JING Pengju,ZHU Dongmei,et al.Effects of dexamethasone on lipopolysaccharide-induced inflammatory damage of human retinal pigment epithelial cells and NLRP3/Caspase-1 pathway[J].Recent Advances in Ophthalmology,2020,40(9):826-830.[doi:10.13389/j.cnki.rao.2020.0188]
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地塞米松对脂多糖诱导人视网膜色素上皮细胞炎症损伤及NLRP3/Caspase-1通路的影响/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
40卷
期数:
2020年9期
页码:
826-830
栏目:
实验研究
出版日期:
2020-09-05

文章信息/Info

Title:
Effects of dexamethasone on lipopolysaccharide-induced inflammatory damage of human retinal pigment epithelial cells and NLRP3/Caspase-1 pathway
作者:
崔铮; 靖鹏举; 朱冬梅; 王媛
450007　河南省郑州市，郑州大学附属郑州中心医院眼科
Author(s):
CUI Zheng; JING Pengju; ZHU Dongmei; WANG Yuan
Department of Ophthalmology，Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou 450007,Henan Province,China
关键词:
地塞米松; 脂多糖; 人视网膜色素上皮细胞; 炎症损伤; 核苷酸结合寡聚化结构域样受体蛋白3/半胱氨酸天冬氨酸蛋白酶-1通路
Keywords:
dexamethasone;  lipopolysaccharide;  human retinal pigment epithelial cells;  inflammatory damage;  nucleotide-binding oligomerization domain like receptor protein 3/Caspase-1 pathway
分类号:
R774.1
DOI:
10.13389/j.cnki.rao.2020.0188
文献标志码:
A
摘要:
目的　探究地塞米松（Dex）对脂多糖（LPS）诱导人视网膜色素上皮（HRPE）细胞炎症损伤及核苷酸结合寡聚化结构域样受体蛋白3（NLRP3）/半胱氨酸天冬氨酸蛋白酶-1（Caspase-1）通路的影响。方法　体外培养ARPE-19细胞株，MMT法检测不同浓度Dex处理细胞24 h、48 h、74 h对LPS诱导ARPE-19细胞存活率的影响，确定Dex最佳处理浓度和时间。ARPE细胞随机分为：正常对照组（NC组）、LPS组（5 mg·L-1 LPS）、Dex组（0.20 mol·L-1 Dex）、Dex+LPS组（0.20 mol·L-1 Dex+5 mg·L-1 LPS），RT-qPCR检测各组细胞肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-1β、IL-6、NLRP3、Caspase-1 mRNA相对表达量，ELISA法测定TNF-α、IL-1β、IL-6蛋白表达水平，Western blot检测NLRP3、Caspase-1蛋白表达水平。结果　同一时间点，与NC组比较，LPS组ARPE-19细胞存活率均显著降低（均为P<0.05），且随着Dex浓度增加，细胞存活率呈先升高后下降的趋势；0.20 mol·L-1 Dex作用LPS诱导的ARPE-19细胞24 h时细胞存活率为(90.77±5.73)%，接近NC组，因此选用0.20 mol·L-1Dex作用24 h用于后续实验。NC组与Dex组间TNF-α、IL-1β、IL-6、NLRP3、Caspase-1 mRNA相对表达量差异均无统计学意义（均为P＞0.05）。与NC组、Dex组比较，LPS组、Dex+LPS组TNF-α、IL-1β、IL-6、 NLRP3、Caspase-1 mRNA及蛋白表达水平均显著升高，且Dex+LPS组TNF-α、IL-1β、IL-6 、NLRP3、Caspase-1 mRNA及蛋白表达水平均显著低于LPS组（均为P<0.05）。结论　Dex可能通过抑制NLRP3/Caspase-1通路减轻LPS诱导的HRPE炎症损伤。
Abstract:
Objective　To investigate the effects of dexamethasone (Dex) on lipopolysaccharide (LPS)-induced inflammatory damage of human retinal pigment epithelial cells (HRPE) and the nucleotide-binding oligomerization domain like receptor protein 3/Caspase-1 (NLRP3/Caspase-1) pathway.Methods　ARPE-19 cell line was cultured in vitro, thiazole blue (MMT) was used to detect the effects of Dex at different concentrations on the activity of LPS-induced ARPE-19 cells to determine the optimum concentration and time. The ARPE-19 cells was divided into four groups: normal control group (NC group, normal growth ARPE-19 cells), LPS inflammatory injury group (LPS group, 5 mg·L-1 LPS treatment to cells), Dex group (0.20 mol·L-1 Dex treatment to cells), Dex + LPS group (0.20 mol·L-1 Dex+ 5 mg·L-1 LPS treatment to cells). The levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6, NLRP3 and Caspase-1 mRNAs were detected by RT-qPCR, the levels of TNF-α, IL-1β, IL-6 were measured by enzyme-linked immunosorbent assay (ELISA), and Western blot was used to detect the expressions of NLRP3 and Caspase-1 proteins in ARPE-19 cells.Results　At the same time, compared with NC group, the activity of ARPE-19 cells in LSP group decreased significantly (all P<0.05), and as the concentration of Dex increases, the cell survival rate increases first and then decreases. The cell survival rate of ARPE-19 cells induced by LPS by 0.20 mol·L-1Dex for 24 hours was (90.77±5.73)%, which was close to the NC group, and therefore, 0.20 mol·L-1 dex for 24 hours was chosen as the optimal concentration for the follow experiment. There was no significant difference in TNF-α, IL-1β, IL-6, NLRP3, Caspase-1 mRNA levels between NC group and Dex group (all P>0.05). Compared with NC group and Dex group, TNF-α, IL-1β, IL-6, NLRP3, Caspase-1 mRNA and protein levels were increased in LPS group and Dex+LPS group, and the levels of TNF-α, IL-1β, IL-6, NLRP3, Caspase1 mRNA and protein in Dex+LPS group were significantly lower than those in LPS group (all P<0.05).Conclusion　Dex may reduce the lipopolysaccharide induced inflammatory damage of HRPE cells by inhibiting NLRP3/Caspase-1 pathway.

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备注/Memo

备注/Memo:
2018年河南省医学科技攻关项目（编号：2018020799）
更新日期/Last Update: 2020-09-05