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缺氧后不同复氧时间对心肌细胞自噬的影响
罗云燕
(昆明医科大学第一附属医院心内科,云南 昆明 650032)
摘要:
[摘要]目的 探讨大鼠心肌细胞(H9C2)不同缺氧复氧(hypoxia reoxygenation,HR)时间对自噬(Autophagy)及细胞活力的影响.方法 体外培养的H9C2细胞按照不同复氧时间随机分为5组,正常对照组(A 组)、缺氧组(B 组)、复氧2 h组(C组)、复氧12 h组(D组)、复氧24 h组(E组).Western Blot检测各组细胞中LC3 II/LC3 I的表达量,MTT法检测各组心肌细胞的活力.结果 HR模型组心肌细胞活力明显低于正常对照组(P<0.05);缺氧组及各复氧组细胞的LC3 II/LC3 I的比值明显高于对照组(P<0.05),其中以12 h组比值最高(P<0.05);缺氧组及各复氧组的细胞活力明显低于对照组(P<0.05),其中以12 h组细胞活力最低(P<0.05).结论 利用三气培养箱可以造成H9C2细胞HR损伤;细胞缺氧可以激活细胞自噬,复氧后自噬进一步激活,以12 h自噬激活最明显,24 h下降至缺氧组水平,同时细胞缺氧造成细胞损伤,复氧后细胞损伤进一步加重,在复氧12 h时细胞活力最低,复氧24 h细胞活力得到改善.
关键词:  [关键词]自噬  大鼠心肌细胞  缺氧复氧
DOI:
分类号:
基金项目:[基金项目]国家自然科学基金资助项目(81560072);云南省科技厅-昆明医科大学联合专项基金资助项目(2013FB128,2014FB037);昆明医科大学2015年硕士研究生创新项目(2015S83)
Effect of Different Reoxygenation Time on the Autophagy of Myocardial Cells
LUO Yun-yan
( Dept. of Cardiology,The 1st Affiliated Hospital of Kunming Medical University,Kunming Yunnan 650032,China )
Abstract:
[Abstract]Objective To investigate the effect of different time of hypoxia reoxygenation (HR) on cell autophagy and cell viability.Methods According to the different reoxygenation time, H9C2 cells were divided into five groups: normal control group(group A), hypoxia group (group B), reoxygen 2h group (group C), reoxygen 12h group(group D)and reoxygen 24h group (group E). The expression of LC3 II/LC3 I in each groups was detected by Western Blot,and the activity of myocardial cells was detected by MTT. Results MTT results showed that the viability of the myocardial cells after hypoxia 2h/reoxygenation 2h was significantly lower than that in the normal control group(P<0.05). The ratio of LC3 II/LC3 I of the cells in group B (hypoxia group), group C (reoxygen 2h group), group D (reoxygen 12h group), group E (reoxygen 24h group) was significantly higher than that of group A (normal control group) (P<0.05). Among them, the ratio of LC3 II/LC3 I of the cells in group C and D were significantly higher than that of group B(P<0.05), paticlularly the group D was the highest (P<0.05) .The activity of myocardial cells in group D was the lowest among all groups. Conclusion Culturing in the three gas incubator can obviously decrease the viability of H9C2cells. Hypoxia can activate autophagy, and autophagy increases significantly after reoxygenation. Autophagy may be activated most obviously at 12 hours and decrease to the level of hypoxia group at 24 hours. Cells can be damaged by hypoxia, and further increase of cellular damage may occur after oxygen inhalation. At 12 hours,the cell viability is the lowest, and the cell viability may be improved at 24h.
Key words:  [Key words]Autophagy  Rat cardiomyocytes  Hypoxia/Reoxygenation