异氟醚调节AMPK/mTOR信号通路减轻大鼠缺氧/复氧所致的心肌细胞损伤OA
Isoflurane alleviates hypoxia/reoxygenation-induced cardiomyocyte injury in rats via regulating AMPK/mTOR signaling pathway
目的 探讨异氟醚调节AMPK/mTOR信号通路减轻大鼠缺氧/复氧(H/R)所致的心肌细胞损伤的作用机制.方法 以不同浓度异氟醚处理H/R诱导的大鼠心肌H9c2细胞,采用MTT法筛选异氟醚的最佳作用浓度.将大鼠心肌H9c2细胞随机分为空白组、H/R组、异氟醚组、化合物C(CC)组、异氟醚+CC组,除空白组外的其余各组进行H/R处理,用异氟醚和AMPK抑制剂CC分组干预后,采用CCK-8法、JC-1荧光染色、流式细胞术分别检测各组细胞活力、线粒体膜电位(∆Ψm)及凋亡率;用透射电镜观察各组H9c2细胞超微结构;用流式细胞术检测各组细胞活性氧(ROS)水平;测定各组H9c2细胞抗氧化酶活性与炎性细胞因子水平;用Western blotting检测各组H9c2细胞AMPK/mTOR信号通路蛋白表达.结果 与空白组相比,H/R组细胞活力、∆Ψm、过氧化氢酶(CAT)与超氧化物歧化酶(SOD)活性水平、p-AMPK/AMPK降低,凋亡率、ROS水平、白细胞介素-6(IL-6)与肿瘤坏死因子α(TNF-α)水平、p-mTOR/mTOR升高(P<0.05).与H/R组相比,异氟醚组细胞活力、∆Ψm升高,凋亡率、ROS水平、IL-6与TNF-α水平、p-mTOR/mTOR降低;CC组细胞各指标变化与异氟醚组相反(P均<0.05).与异氟醚组相比,异氟醚+CC组细胞活力、∆Ψm降低,凋亡率、ROS水平、IL-6与TNF-α水平、p-mTOR/mTOR升高(P均<0.05).结论 异氟醚可通过激活AMPK/mTOR信号通路减轻H/R心肌细胞损伤.
Objective To investigate the mechanisms through which isoflurane alleviates hypoxia/reoxygenation(H/R)injury to cardio-myocytes via regulating the AMPK/mTOR signaling pathway.Methods Using H/R-induced rat cardiomyocytes,H9c2 cells were treated with different isoflurane concentrations,and the optimal isoflurane concentration was determined using an MTT assay.H9c2 cells were sto-chastically assigned to control,H/R,isoflurane,compound C(CC),and isoflurane+CC groups.All groups other than the control group were subjected to H/R treatment.After treatment with isoflurane and the AMPK inhibitor CC,the cell viability,mitochondrial membrane poten-tial(∆Ψm),and apoptosis rate were detected in each group using the CCK-8 method,JC-1 fluorescence staining,and flow cytometry,respectively.Transmission electron microscopy and flow cytometry were used to observe the ultrastructure of the H9c2 cells and detect the levels of reactive oxygen species(ROS)in the cells in each group,respectively.The antioxidant enzyme activity and levels of inflammatory factors released in the H9c2 cells were measured in all groups.Western blotting was used to detect the expression levels of AMPK/mTOR pathway proteins in the H9c2 cells in each group.Results The cell viability,∆Ψm,catalase(CAT)and superoxide dismutase(SOD)activities,and p-AMPK/AMPK were lower in H/R group than in the control group,whereas the apoptosis rate;ROS,interleukin-6(IL-6),and tumor necrosis factor α(TNF-α)levels;and p-mTOR/mTOR were higher in the H/R group than in the control group(all P<0.05).The cell viability and ∆Ψm were higher in the isoflurane group than in the H/R group.The apoptosis rate;ROS,IL-6,and TNF-α levels;and p-mTOR/mTOR were lower the isoflurane group than in the H/R group.The changes in the various parameters in the cells in the CC group were the opposite to those in isoflurane group(all P<0.05).The cell viability and ∆Ψm were lower in the isoflurane+CC group compared with those in the isoflurane group,whereas the apoptosis rate;ROS,IL-6,and TNF-α levels;and p-mTOR/mTOR were higher in the isoflurane+CC group(all P<0.05).Conclusion Isoflurane alleviates H/R-induced cardiomyocyte injury via activating the AMPK/mTOR signaling pathway.
陈艳林;张晓敏;刘娜;袁慧敏;邢珍;姚杰
河北北方学院附属第一医院麻醉科,河北 张家口 075061河北北方学院附属第一医院麻醉科,河北 张家口 075061河北北方学院附属第一医院麻醉科,河北 张家口 075061河北北方学院附属第一医院麻醉科,河北 张家口 075061河北北方学院附属第一医院麻醉科,河北 张家口 075061河北北方学院附属第一医院麻醉科,河北 张家口 075061
医药卫生
异氟醚AMPK/mTOR缺氧/复氧心肌细胞保护作用
isofluraneAMPK/mTORhypoxia/reoxygenationcardiomyocytesprotective function
《中国医科大学学报》 2026 (3)
259-264,271,7
河北省医学科学研究课题(20241120)
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