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| 稳心颗粒调控SIRT1/PGC-1α/PPARs改善H9c2细胞缺氧模型能量代谢及Cx43的机制研究 |
| Mechanism of Wenxin granule regulating SIRT1/PGC-1α/PPARs to improve energy metabolism and Cx43 in H9c2 cardiomyocytes under hypoxiaYang Ding, Cui Xiyuan, Wang Zhe, Lou Lixia, Nie Bo, Zhao Jiuli, Zhao Mingjing, Wu Aiming * |
| 投稿时间：2024-01-30 录用日期：2024-04-22 |
| DOI： |
| 中文关键词: 稳心颗粒 心肌细胞 能量代谢 缝隙连接 SIRT1/PGC-1α/PPARs |
| 英文关键词: Wenxin granule Cardiomyocytes Energy metabolism Gap junction SIRT1/PGC-1α/PPARs |
| 基金项目:国家自然科学基金面上项目(82274449);北京中医药大学东直门医院2022科技创新专项(DZMKJCX-2022-008) |
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| 中文摘要: |
| 摘要：目的：研究稳心颗粒调控SIRT1/PGC-1α/PPARs改善缺氧条件下H9c2细胞能量代谢以及对Cx43的影响。方法：使用1%O2、5%CO2、94%N2培养箱构建H9c2细胞缺氧损伤模型，随机分为Control组、Hypoxia组、WXKL组、TMZ组。采用CCK-8法检测细胞活力；Elisa法检测各组细胞ATP，ADP和AMP含量，并计算ADP/ATP、AMP/ATP的比值；免疫荧光检测Cx43蛋白表达和分布变化；蛋白印迹法检测Cx43、p-Cx43，信号通路SIRT1/PGC-1α/PPARs相关蛋白以及能量代谢相关蛋白CD36、CPT-1α、GLUT4。结果：与Control组相比，Hypoxia组细胞活力明显降低（P＜0.01），ATP含量下降（P＜0.01），ADP、AMP上升（P＜0.01），ADP/ATP、AMP/ATP比值升高（P＜0.01），Cx43阳性细胞表达减少，心肌细胞内Cx43、p-Cx43、SIRT1、PGC-1α、PPARα、PPARδ、CD36、CPT-1α、GLUT4蛋白表达下降（P＜0.05）；与Hypoxia组比较，WXKL组和TMZ组细胞活力明显上升（P＜0.01），ATP含量升高（P＜0.05）、ADP、AMP水平下降（P＜0.01）、ADP/ATP、AMP/ATP比值下降（P＜0.01），Cx43阳性细胞荧光表达增多，心肌细胞内Cx43、p-Cx43、SIRT1、PGC-1α、PPARα、PPARδ、CD36、CPT-1α、GLUT4蛋白表达升高（P＜0.05），TMZ组p-Cx43和CPT-1α蛋白表达无明显变化。结论：稳心颗粒可以改善缺氧条件下H9c2心肌细胞能量代谢障碍，保护Cx43，其机制与调节SIRT1/PGC-1α/PPARs信号通路有关。 |
| 英文摘要: |
| Abstract: Objective: To study the effects of Wenxin Granule on the regulation of SIRT1/PGC-1α/PPARs in improving the energy metabolism of H9c2 cells under hypoxic conditions and its influence on Cx43. Methods: A hypoxic injury model of H9c2 cardiomyocytes was constructed using a culture box with 1% O2, 5% CO2, and 94% N2. Cells were randomly divided into Control group, Hypoxia group, WXKL group, and TMZ group. Cell viability was measured by CCK-8 assay; ATP, ADP, and AMP content in cells of each group were detected by Elisa, and the ratios of ADP/ATP and AMP/ATP were calculated; Immunofluorescence staining was used to detect the expression and distribution changes of Cx43 protein; Western blot was employed to measure the expression of Cx43, p-Cx43, SIRT1/PGC-1α/PPARs signaling pathway-related proteins, and energy metabolism-related proteins such as CD36, CPT-1α, and GLUT4. Results: Compared with the Control group, the Hypoxia group showed a significant decrease in cell viability (P<0.01), a decrease in intracellular ATP (P<0.01), an increase in ADP and AMP (P<0.01), and elevated ratios of ADP/ATP and AMP/ATP (P<0.01). The expression of positive Cx43 cells was reduced, and the expression of intracellular Cx43, p-Cx43, SIRT1, PGC-1α, PPARα, PPARδ, CD36, CPT-1α, and GLUT4 proteins was downregulated (P<0.05). Compared with the Hypoxia group, the cell viability in the WXKL and TMZ groups significantly increased (P<0.01), ATP content rose (P<0.05), levels of ADP and AMP decreased (P<0.01), and the ratios of ADP/ATP and AMP/ATP decreased (P<0.01). The fluorescence expression of positive Cx43 cells increased, and the expression of intracellular Cx43, p-Cx43, SIRT1, PGC-1α, PPARα, PPARδ, CD36, CPT-1α, and GLUT4 proteins was upregulated (P<0.05). There was no significant change in the expression of p-Cx43 and CPT-1α proteins in the TMZ group. Conclusion: Wenxin Granule can improve the energy metabolism disorder of H9c2 cardiomyocytes under hypoxic conditions and protect Cx43. The mechanism is related to the regulation of the SIRT1/PGC-1α/PPARs signaling pathway. |
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