| 耿小晶,董 军,曾 幸,曾庆敏,孙卫宁,薛 青,张少玲.Nox2在AngII诱导的H295R细胞醛固酮合成中的作用[J].,2023,(23):4401-4406 |
| Nox2在AngII诱导的H295R细胞醛固酮合成中的作用 |
| Effects of Nox2 on AngII-stimulated Aldosterone Biosynthesis in H295R Cells |
| 投稿时间：2023-05-30 修订日期：2023-06-23 |
| DOI：10.13241/j.cnki.pmb.2023.23.001 |
| 中文关键词: NADPH氧化酶 Nox2 醛固酮合成 |
| 英文关键词: NADPH oxidase Nox2 Aldosterone biosynthesis |
| 基金项目:国家自然科学基金项目（81600606）；广东省自然科学基金资助项目（2014A030313081） |
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| 中文摘要: |
| 摘要 目的：观察Nox2对AngII活化的人肾上腺皮质腺癌细胞（H295R细胞）醛固酮合成的影响。方法：将H295R细胞分为正常对照组、AngII、AngII＋gp91ds-tat（Nox2抑制剂）组、AngII+PEG-Cat（H2O2清除剂）组、AngII+Nox2 siRNA组及不同时间的AngII组，采用Q-PCR和western blot检测醛固酮合成酶（CYP11B2）和Nox2基因及蛋白水平；放免法检测细胞上清液醛固酮浓度；应用流式细胞术和酶标仪检测细胞内Nox2来源的ROS和H2O2的含量。结果：10 nmol/L AngII以时间依赖性增加H295R细胞内ROS和H2O2含量、Nox2和CYP11B2表达、醛固酮合成（P<0.05）。与正常对照组相比，gp91ds-tat和PEG-Cat明显降低AngII诱导的细胞内ROS和H2O2含量（P<0.05），而gp91ds-tat组和PEG-Cat组AngII诱导的细胞内ROS和H2O2抑制作用无差别（P>0.05）。10 nmol/L AngII 处理24 h诱导H295R细胞CYP11B2表达（P<0.05），而gp91ds-tat组、PEG-Cat组和Nox2 siRNA组明显抑制AngII诱导H295R细胞CYP11B2表达（P<0.05）。结论：Nox2来源的ROS在AngII诱导的醛固酮合成过程中起主要调控作用。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the role of Nox2 on AngII-stimulated aldosterone biosynthesis. Methods: H295R cells were cultured and randomly divided into control group, AngII group, AngII＋gp91ds-tat group, AngII+PEG-Cat group, AngII+Nox2 siRNA group. Q-PCR and western blot were performed to determine CYP11B2 and Nox2 expression in H295R cells. Aldosterone level in cells culture supernatant was determined by radioimmunoassay. Nox2-derived ROS and H2O2 production in H295R cells were determined by flow cytometric analysis and fluorescence microplate reader. Results: 10 nmol/L AngII increased cellular ROS and H2O2 generation, increased the Nox2 and CYP11B2 expression and accelerated aldosterone biosynthesis in a time-dependent manner (P<0.05). And AngII-induced cellular ROS and H2O2 production in both AngII+gp91ds-tat and AngII+PEG-Cat groups were lower than those in AngII group (P<0.05)．However, there were no difference in inhibition of cellular ROS and H2O2 between treatment with gp91ds-tat and PEG-Cat (P>0.05). The expression of CYP11B2 in H295R cells was up-regulated after treated with 10 nmol/L AngII for 24 h (P<0.05), while gp91ds-tat, PEG-Cat and Nox2 siRNA significantly abrogated the CYP11B2 expression in H295R cells (P<0.05). Conclusion: Nox2 dependent ROS play an important role in aldosterone synthesis. |
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