| 李 均,徐 艺,冉 阳,徐 刚,王均生.汉黄芩素调节AMPK/NLRP3信号通路对H/R诱导的心肌细胞凋亡的影响[J].,2024,(17):3211-3216 |
| 汉黄芩素调节AMPK/NLRP3信号通路对H/R诱导的心肌细胞凋亡的影响 |
| Effect of Wogonin on H/R Induced Cardiomyocyte Apoptosis by Regulating the AMPK/NLRP3 Signaling Pathway |
| 投稿时间：2024-02-28 修订日期：2024-03-23 |
| DOI：10.13241/j.cnki.pmb.2024.17.003 |
| 中文关键词: 黄芩素 AMPK/NLRP3信号通路 心肌细胞 细胞凋亡 |
| 英文关键词: Wogonin AMPK/NLRP3 signaling pathway Cardiomyocyte Apoptosis |
| 基金项目:重庆市自然科学基金面上项目（cstc2021jcyj-msxmX1061） |
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| 中文摘要: |
| 摘要 目的：探讨汉黄芩素（WOG）调节单磷酸腺苷活化蛋白激酶（AMPK）/NOD样受体蛋白3（NLRP3）信号通路对缺氧/再氧合（H/R）诱导的心肌细胞凋亡的影响。方法：将H9C2细胞分为Control组（正常培养）、H/R组（H/R诱导）、L（低剂量）-WOG组、M（中剂量）-WOG组、H（高剂量）-WOG组（在H/R诱导的基础上分别加入40、80、120 μmol/L的WOG）、WOG+Compound C组（在H-WOG组基础上加入10 μmol/L AMPK抑制剂Compound C）。噻唑蓝（MTT）法和5-乙炔基-2′脱氧尿嘧啶核苷（EdU）染色检测WOG对H9C2细胞增殖的影响；流式细胞术检测WOG对H9C2细胞凋亡的影响；酶联免疫吸附试验（ELISA）检测H9C2细胞血清氧化应激指标[丙二醛（MDA）、活性氧类物质（ROS）、超氧化物歧化酶（SOD）]和炎症因子[白介素（IL）-1β、IL-18]水平；蛋白免疫印迹（WB）法检测H9C2细胞AMPK、NLRP3、B细胞淋巴瘤-2（Bcl-2）、Bcl-2相关X蛋白（Bax）蛋白表达。结果：H/R组H9C2细胞的光密度值（OD490）、EdU阳性细胞率、SOD、AMPK、Bcl-2低于Control组，细胞凋亡率、IL-1β、IL-18、ROS、MDA、NLRP3、Bax高于Control组（P<0.05）；与H/R组比较，L-WOG组、M-WOG组、H-WOG组OD490、EdU阳性细胞率、SOD、AMPK、Bcl-2表达升高，细胞凋亡率、IL-1β、IL-18、ROS、MDA、NLRP3、Bax降低（P<0.05）；WOG+Compound C组OD490、EdU阳性细胞率、SOD、AMPK、Bcl-2低于H-WOG组，细胞凋亡率、IL-1β、IL-18、ROS、MDA、NLRP3、Bax表达高于H-WOG组（P<0.05）。结论：WOG可以抑制H/R诱导的心肌细胞凋亡，其机制可能是通过介导AMPK/NLRP3信号通路有关。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effect of wogonin (WOG) on hypoxia/reoxygenation (H/R)-induced cardiomyocyte apoptosis by regulating adenosine monophosphate-activated protein kinase (AMPK)/NOD-like receptor protein 3 (NLRP3) signaling pathway. Methods: H9C2 cells were divided into Control group (normal culture), H/R group (H/R induction), L (low dose)-WOG group, M(medium dose)-WOG group, H(high dose)-WOG group (40, 80, 120 μmol/L WOG were added on the basis of H/R induction), WOG+Compound C group (10 μmol/L AMPK inhibitor Compound C was added on the basis of H-WOG group). The effect of WOG on the proliferation of H9C2 cells was detected by methyl thiazolyl tetrazolium (MTT) assay and 5-ethynyl-2 ' -deoxyuridine (EdU) staining. The effect of WOG on apoptosis of H9C2 cells was detected by flow cytometry. The levels of serum oxidative stress indexes [malondialdehyde (MDA), reactive oxygen species (ROS), superoxide dismutase (SOD)] and inflammatory factors [interleukin (IL)-1β, IL-18] in H9C2 cells were detected by enzyme-linked immunosorbent assay (ELISA). The expression of AMPK, NLRP3, B cell lymphoma-2 (Bcl-2) and Bcl-2 associated X protein (Bax) in H9C2 cells was detected by Western blot (WB). Results: The optical density (OD490), EdU positive cell rate, SOD, AMPK and Bcl-2 of H9C2 cells in H/R group were lower than those in Control group, and the apoptosis rate, IL-1β, IL-18, ROS, MDA, NLRP3 and Bax were higher than those in Control group (P<0.05). Compared with H/R group, OD490, EdU positive cell rate, SOD, AMPK and Bcl-2 expression in L-WOG group, M-WOG group and H-WOG group increased, while apoptosis rate, IL-1β, IL-18, ROS, MDA, NLRP3 and Bax decreased (P<0.05). The OD490, EdU positive cell rate, SOD, AMPK and Bcl-2 in WOG+Compound C group were lower than those in H-WOG group, and the apoptosis rate, IL-1β, IL-18, ROS, MDA, NLRP3 and Bax expression were higher than those in H-WOG group (P<0.05). Conclusion: WOG can inhibit H/R-induced cardiomyocyte apoptosis, which may be relate to the AMPK/NLRP3 signaling pathway. |
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