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.
Objective To investigate the effect of different time of hypoxia reoxygenation(HR)on cellautophagy and cell viability.Methods According to the different reoxygenation time,H9C2cells were dividedinto five groups:normal control group(group A),hypoxia group(group B),reoxygen2h group(group C),reoxygen12h group(group D)and reoxygen24h group(group E).The expression of LC3II/LC3I in each groupswas detected by Western Blot,and the activity of myocardial cells was detected by MTT.Results MTT resultsshowed that the viability of the myocardial cells after hypoxia2h/reoxygenation2h was significantly lower than that inthe normal control group(P<0.05).The ratio of LC3II/LC3I of the cells in group B(hypoxia group),group C(reoxygen2h group),group D(reoxygen12h group),group E(reoxygen24h group)was significantly higherthan that of group A(normal control group)(P<0.05).Among them,the ratio of LC3II/LC3I of the cells ingroup 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 inthe three gas incubator can obviously decrease the viability of H9C2cells.Hypoxia can activate autophagy,andautophagy increases significantly after reoxygenation.Autophagy may be activated most obviously at12hours and decrease to the level of hypoxia group at24hours.Cells can be damaged by hypoxia,and further increase of cellulardamage may occur after oxygen inhalation.At12hours,the cell viability is the lowest,and the cell viability may beimproved at24h.
作者
罗云燕
胡钊
肖践明
蔡红雁
李琳
郭涛
LUO Yun-yan;HU Zhao;XIAO Jian-ming;CAI Hong-yan;LI Lin;GUO Tao(Dept. of Cardiology,The 1st Affiliated Hospital of Kunming Medical University,Kunming Yunnan 650032,China)
出处
《昆明医科大学学报》
CAS
2016年第10期5-9,共5页
Journal of Kunming Medical University
基金
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)
关键词
自噬
大鼠心肌细胞
缺氧复氧
Autophagy
Rat cardiomyocytes
Hypoxia/Reoxygenation
