| 谢孝平,沈小艳,李博文,刘天宇,王志维.芦丁通过激活NRF2/HO-1通路抑制Ang II诱导的血管平滑肌细胞表型转化和炎症反应[J].,2023,(22):4201-4206 |
| 芦丁通过激活NRF2/HO-1通路抑制Ang II诱导的血管平滑肌细胞表型转化和炎症反应 |
| Rutin Inhibits Ang II-induced Vascular Smooth Muscle Cell Phenotypic Switching and Inflammatory Response through Activating NRF2/HO-1 Pathway |
| 投稿时间：2023-05-25 修订日期：2023-06-11 |
| DOI：10.13241/j.cnki.pmb.2023.22.001 |
| 中文关键词: 芦丁 血管平滑肌细胞 表型转化 炎症反应 氧化应激 |
| 英文关键词: Rutin Vascular Smooth Muscle Cells Phenotypic Switching Inflammatory Response Oxidative Stress |
| 基金项目:国家自然科学基金项目（82070481；82200522） |
|
| 摘要点击次数: 669 |
| 全文下载次数: 406 |
| 中文摘要: |
| 摘要 目的：探讨芦丁（Rutin）对血管紧张素II（Angiotensin II，Ang II）诱导的小鼠主动脉血管平滑肌细胞表型转化和炎症反应的影响及机制。方法：采用Ang II处理小鼠主动脉平滑肌细胞构建表型转化和炎症反应模型。将对数生长期的小鼠主动脉血管平滑肌细胞分为以下4组：正常对照组（Control组），Rutin组（100 μM），Ang II组（1μM），Rutin+ Ang II组（100 μM，1 μM）。采用细胞增殖实验（Cell Counting Kit-8，CCK8）检测芦丁对小鼠主动脉平滑肌细胞活力的影响，采用蛋白免疫印迹实验检测α平滑肌肌动蛋白（α-Smooth muscle actin，α-SMA）、平滑肌蛋白22α（Smooth muscle protein 22-alpha，SM22α）、骨桥蛋白（Osteopontin，OPN）、基质金属蛋白酶2（Matrix metalloproteinase 2，MMP2）、基质金属蛋白酶9（Matrix metalloproteinase 9，MMP9）、白细胞介素6（Interleukin 6，IL6）、肿瘤坏死因子-α（Tumor necrosis factor-alpha，TNF-α）、核因子E2相关因子2（nuclear factor erythroid 2-related factor 2，Nrf2）、血红素氧合酶-1（Heme oxygenase-1，HO-1）、核因子活化B细胞κ轻链增强子（nuclear factor kappa-light-chain-enhancer of activated B cells，NF-κB）的蛋白表达和磷酸化水平，采用双抗体一步夹心法酶联免疫吸附试验（Enzyme-linked immunosorbent assay，ELISA）检测细胞上清中TNF-α和IL6细胞因子水平，采用荧光显微镜和流式细胞术检测小鼠主动脉血管平滑肌细胞活性氧水平。结果：与对照组相比，Ang II组收缩型标志物α-SMA和SM22α表达水平降低、合成型标志物OPN表达水平升高，炎症相关因子MMP2、MMP9、IL6和TNF-α表达水平升高，NRF2和HO-1表达水平降低，NRF2及NF-κB的磷酸化增加。此外，相较于Ang II组，Rutin+ Ang II组收缩型标志物α-SMA和SM22α表达水平升高、合成型标志物OPN表达水平降低，炎症相关因子MMP2、MMP9、IL6和TNF-α表达水平降低，NRF2和HO-1表达水平升高，NRF2及NF-κB的磷酸化水平降低。结论：芦丁可以抑制Ang II诱导的小鼠主动脉血管平滑肌细胞表型转化和炎症反应，可能与其激活NRF2/HO-1通路和抑制活性氧的产生有关。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effects and mechanisms of rutin on angiotensin II (Ang II)-induced phenotype transition and inflammatory response in mouse aortic vascular smooth muscle cells. Methods: Mouse aortic vascular smooth muscle cells in logarithmic growth phase were divided into four groups: Control group (Control), Rutin group (100 μM), Ang II group (1 μM), and Rutin+Ang II group (100 μM, 1 μM). Cell viability was assessed using the Cell Counting Kit-8 (CCK-8) assay to evaluate the effect of rutin on mouse aortic vascular smooth muscle cell viability. Protein immunoblotting was performed to examine the expression and phosphorylation levels of α-smooth muscle actin (α-SMA), Smooth muscle protein 22-alpha (SM22α), Osteopontin (OPN), Matrix metalloproteinase 2 (MMP2), Matrix metalloproteinase 9 (MMP9), Interleukin 6 (IL6), Tumor necrosis factor-alpha (TNF-α), Nuclear factor erythroid 2-related factor 2 (NRf2), Heme oxygenase-1 (HO-1), and Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB). Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of TNF-α and IL6 in the cell supernatant. Fluorescence microscopy and flow cytometry were employed to detect the levels of reactive oxygen species (ROS) in mouse aortic vascular smooth muscle cells. Results: Compared to the control group, the Ang II group exhibited decreased expression levels of contractile markers α-SMA and SM22α, increased expression level of synthetic marker OPN, elevated expression levels of inflammatory factors MMP2, MMP9, IL6, and TNF-α, decreased expression levels of NRf2 and HO-1, and increased phosphorylation levels of NRf2 and NF-κB. Additionally, compared to the Ang II group, the Rutin+Ang II group showed increased expression levels of contractile markers α-SMA and SM22α, decreased expression level of synthetic marker OPN, reduced expression levels of inflammatory factors MMP2, MMP9, IL6, and TNF-α, increased expression levels of NRf2 and HO-1, and decreased phosphorylation levels of NRf2 and NF-κB. Conclusion: Rutin can inhibit Ang II-induced phenotype transition and inflammatory response in mouse aortic vascular smooth muscle cells, possibly through activation of the NRF2/HO-1 pathway and inhibition of ROS production. |
|
查看全文
查看/发表评论 下载PDF阅读器 |
| 关闭 |
|
|
|
