VitaePro抑制线粒体通路对心肌细胞缺氧损伤的保护作用

目的：探讨VitaePro对心肌细胞缺氧损伤的保护作用及其机制。方法将大鼠心肌细胞H9c2分为四组：正常对照组(H9c2)、缺氧损伤模型组(Hypoxia+H9c2)、红花油组(Safflower oil+hypoxia+H9c2)和VitaePro组(VitaePro+hypoxia+H9c2)。MTT和流式细胞术分别检测心肌细胞增殖和凋亡情况；Western blot检测线粒体凋亡通路相关蛋白的表达，包括B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关蛋白X (BAX)、凋亡蛋白-3(Caspase-3)和裂解后的Caspase-3(Cleaved caspase-3)；试剂盒检测各组心肌细胞硫代巴比妥酸反应物(TBARS)水平、丙二醛（MDA）含量和超氧化物歧化酶(SOD)的活性。最后，Western blot检测心血管损伤标志蛋白的表达，包括心肌肌钙蛋白T (cTnT)、肌红蛋白(Mb)、肌酸激酶同工酶MB (CK-MB)和乳酸脱氢酶(LDH)。结果缺氧损伤模型组与对照组比较，细胞增殖显著下降，凋亡率显著增高(4.2%~14.6%，P<0.05)，Bcl-2蛋白的表达明显降低，Bax和Cleaved caspase-3的表达显著升高(P<0.05)，TBARS水平和MDA含量升高(P<0.05)，SOD活性从44.9 U/mg降低到13.2 U/mg，心血管损伤标志蛋白cTnT、Mb、CK-MB和LDH的表达显著升高(P<0.05)。VitaePro组与缺氧组相比，细胞的增殖升高(P<0.05)，凋亡率降低到6.4%，Bcl-2的表达升高，Bax和Cleaved caspase-3的表达明显降低(P<0.05)，TBARS水平和MDA含量明显降低(P<0.05)，SOD活性增大到41.0 U/mg，cTnT、Mb、CK-MB和LDH的表达显著降低(P<0.05)。结论 VitaePro可以通过抑制线粒体凋亡通路和通过抗氧化作用减轻缺氧诱导的大鼠心肌细胞损伤。

Protective effects of VitaePro inhibiting the mitochondrial pathways on hypoxia-induced cardiomyocytes injury

Objective To study the protective effect and mechanism of VitaePro on hypoxia-induced cardiomy-ocytes injury. Methods Rat myocardial cells H9c2 were randomly divided into four groups:H9c2 group (the control group), Hypoxia+H9c2 group, Safflower oil+hypoxia+H9c2 group and VitaePro+hypoxia+H9c2 group. Cell prolifera-tion and cell apoptosis were detected by MTT and flow cytometry, respectively. The expression levels of mitochondrial apoptotic pathway related proteins were measured by Western blot, including B cell lymphoma 2 (Bcl-2), Bcl-2-associat-ed X (Bax), Caspase-3 and Cleaved caspase-3. The thiobarbituric acid reactive substance (TBARS) level, malondialde-hyde (MDA) content and superoxide dismutase (SOD) activity were measured by corresponding kits. Finally, the expres-sion of cardiovascular injury markers protein were measured by Western blot, including cardiac troponin T (cTnT), myo-hemoglobin (Mb), creatine kinase isozyme MB (CK-MB) and lactic dehydrogenase (LDH). Results Compared with control group, the cell proliferation in Hypoxia+H9c2 group was significantly decreased (P<0.05), and the cell apoptosis was significantly increased (4.2%~14.6%, P<0.05). The expression level of Bcl-2 was significantly decreased, while Bax and Cleaved caspase-3 were significantly increased (P<0.05). The level of TBARS and the content of MDA were in-creased, and the activity of SOD was decreased from 44.9 U/mg to 13.2 U/mg. The expressions of cardiovascular injury markers cTnT, Mb, CK-MB and LDH were significantly increased (P<0.05). Compared with Hypoxia+H9c2 group, the cell proliferation in VitaePro+hypoxia+H9c2 group was increased (P<0.05), and the cell apoptosis was decreased to 6.4%. The expression level of Bcl-2 was increased, while Bax and Cleaved caspase-3 were significantly decreased (P<0.05). The level of TBARS and the content of MDA was significantly decreased (P<0.05), and the activity of SOD was increased to 41.0 U/mg. The expressions of cardiovascular injury markers cTnT, Mb, CK-MB and LDH were significant-ly decreased (P<0.05). Conclusion VitaePro can reduce hypoxia-induced myocardial cell injury in rats via inhibiting mitochondrial apoptosis pathway and through antioxidant effect.