在严重哮喘中Nrf2可通过靶向miR-29b调控下游基因，以及葡萄籽原花青素提取物在激素不敏感哮喘小鼠模型中的作用
2022/05/17
摘要
背景:葡萄籽原花青素提取物(GSPE)对治疗严重哮喘(SA)有效。
目的：检测Nrf2-miR-29b轴与SA的关系，检测预防性使用GSPE是否通过其缓解SA。
方法：2017年2月至12月，我们招募了10名健康对照、10名非严重哮喘(nSA)患者和9名急性哮喘患者。提取这些志愿者的外周血单个核细胞。6周龄雌性BALB/c小鼠经OVA、Al(OH)3和LPS刺激31天，建立激素不敏感哮喘小鼠模型。给药组注射地塞米松(5 mg/kg/d)，加或不加GSPE(100mg/kg/d)。对照组给予PBS。我们对动物和细胞模型进行实时荧光定量PCR、western blot和荧光素酶报告检测。
结果：SA组Nrf2蛋白、Nrf2 mRNA和miR-29b的浓度均显著低于nSA组和对照组。相反，与其他两组相比，SA中血小板衍生生长因子C (PDGFC)、磷酸肌苷-3-激酶调节亚基1 (PIK3R1)和型胶原蛋白α1(COL3A1)的水平更高。PDGFC、PIK3R1、COL3A1是miR-29b的靶基因。与OVA+LPS相比，GSPE+DXM显著提高Nrf2(+188%)、Nrf2 mRNA(+506%)和miR-29b(+201%)的表达，显著降低PDGFC(-72%)、PIK3R1(-40%)和COL3A1(-65%)的表达。
结论：Nrf2-miR-29b轴参与SA的发病机制。GSPE作为一种辅助药物，可能是一种潜在的SA治疗药物。
Nrf2 regulates downstream genes by targeting miR-29b in severe asthma and the role of grape seed proanthocyanidin extract in a murine model of steroid-insensitive asthma
Y. Qian, Y. Sun, Y. Chen, Z. Mao, Y. Shi, D. Wu, et al.
Abstract
BACKGROUND：Grape seed proanthocyanidin extract (GSPE) is effective in treating severe asthma (SA).
OBJECTIVE：To examine the relationship between Nrf2-miR-29b axis and SA, and to detect whether preventive use of GSPE relieves SA via it.
METHODS：We recruited 10 healthy controls, 10 patients with non-severe asthma (nSA), and 9 patients with SA from February 2017 to December 2017. Peripheral blood mononuclear cells from these volunteers were extracted. A murine model of steroid-insensitive asthma was established in six-week-old female BALB/c mice that were sensitised and challenged with OVA, Al(OH)3 and LPS for 31 days. Mice in the treated groups were injected with DXM (5 mg/kg/d), with or without GSPE (100 mg/kg/d). Control group received PBS. We performed quantitative real-time PCR, western blot and luciferase reporter assay in animal and cell models.
RESULTS：SA group demonstrated significantly lower concentrations of Nrf2 protein, Nrf2 mRNA, and miR-29b than nSA group and control group. Conversely, higher levels of platelet derived growth factor C (PDGFC), phosphoinositide-3-kinase regulatory subunit 1 (PIK3R1), and collagen type III alpha 1 (COL3A1) were measured in SA than in the other two groups. PDGFC, PIK3R1, and COL3A1 were the target genes of miR-29b. GSPE + DXM significantly elevated the expression of Nrf2 (+188%), Nrf2 mRNA (+506%), and miR-29b (+201%), and significantly reduced the expression of PDGFC (−72%), PIK3R1 (−40%), and COL3A1 (−65%) compared with OVA + LPS.
CONCLUSIONS：Nrf2-miR-29b axis is involved in the pathogenesis of SA. GSPE, as an adjuvant drug, maybe a potential therapeutic agent for SA.
背景:葡萄籽原花青素提取物(GSPE)对治疗严重哮喘(SA)有效。
目的：检测Nrf2-miR-29b轴与SA的关系，检测预防性使用GSPE是否通过其缓解SA。
方法：2017年2月至12月，我们招募了10名健康对照、10名非严重哮喘(nSA)患者和9名急性哮喘患者。提取这些志愿者的外周血单个核细胞。6周龄雌性BALB/c小鼠经OVA、Al(OH)3和LPS刺激31天，建立激素不敏感哮喘小鼠模型。给药组注射地塞米松(5 mg/kg/d)，加或不加GSPE(100mg/kg/d)。对照组给予PBS。我们对动物和细胞模型进行实时荧光定量PCR、western blot和荧光素酶报告检测。
结果：SA组Nrf2蛋白、Nrf2 mRNA和miR-29b的浓度均显著低于nSA组和对照组。相反，与其他两组相比，SA中血小板衍生生长因子C (PDGFC)、磷酸肌苷-3-激酶调节亚基1 (PIK3R1)和型胶原蛋白α1(COL3A1)的水平更高。PDGFC、PIK3R1、COL3A1是miR-29b的靶基因。与OVA+LPS相比，GSPE+DXM显著提高Nrf2(+188%)、Nrf2 mRNA(+506%)和miR-29b(+201%)的表达，显著降低PDGFC(-72%)、PIK3R1(-40%)和COL3A1(-65%)的表达。
结论：Nrf2-miR-29b轴参与SA的发病机制。GSPE作为一种辅助药物，可能是一种潜在的SA治疗药物。
（中日友好医院呼吸与危重症医学科 李春晓 摘译 林江涛 审校）
（Pharm Biol. 2022; 60(1): 347–358.doi: 10.1080/13880209.2022.2032205）
（Pharm Biol. 2022; 60(1): 347–358.doi: 10.1080/13880209.2022.2032205）
Nrf2 regulates downstream genes by targeting miR-29b in severe asthma and the role of grape seed proanthocyanidin extract in a murine model of steroid-insensitive asthma
Y. Qian, Y. Sun, Y. Chen, Z. Mao, Y. Shi, D. Wu, et al.
Abstract
BACKGROUND：Grape seed proanthocyanidin extract (GSPE) is effective in treating severe asthma (SA).
OBJECTIVE：To examine the relationship between Nrf2-miR-29b axis and SA, and to detect whether preventive use of GSPE relieves SA via it.
METHODS：We recruited 10 healthy controls, 10 patients with non-severe asthma (nSA), and 9 patients with SA from February 2017 to December 2017. Peripheral blood mononuclear cells from these volunteers were extracted. A murine model of steroid-insensitive asthma was established in six-week-old female BALB/c mice that were sensitised and challenged with OVA, Al(OH)3 and LPS for 31 days. Mice in the treated groups were injected with DXM (5 mg/kg/d), with or without GSPE (100 mg/kg/d). Control group received PBS. We performed quantitative real-time PCR, western blot and luciferase reporter assay in animal and cell models.
RESULTS：SA group demonstrated significantly lower concentrations of Nrf2 protein, Nrf2 mRNA, and miR-29b than nSA group and control group. Conversely, higher levels of platelet derived growth factor C (PDGFC), phosphoinositide-3-kinase regulatory subunit 1 (PIK3R1), and collagen type III alpha 1 (COL3A1) were measured in SA than in the other two groups. PDGFC, PIK3R1, and COL3A1 were the target genes of miR-29b. GSPE + DXM significantly elevated the expression of Nrf2 (+188%), Nrf2 mRNA (+506%), and miR-29b (+201%), and significantly reduced the expression of PDGFC (−72%), PIK3R1 (−40%), and COL3A1 (−65%) compared with OVA + LPS.
CONCLUSIONS：Nrf2-miR-29b axis is involved in the pathogenesis of SA. GSPE, as an adjuvant drug, maybe a potential therapeutic agent for SA.
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