摘要
目的:构建体外小鼠成肌细胞间歇性低氧/复氧模型并验证。方法:设定三气培养箱的混合气中1%或4%O2含量为低氧培养条件,二氧化碳培养箱中的条件为复氧条件,将小鼠成肌细胞(C2C12)分别按不同氧浓度(1%和4%)及复氧循环周期(6 h和8 h)处理,结束后观察细胞增殖情况,测定细胞培养液中葡萄糖、乳酸脱氢酶(lactic dehydrogenase, LDH)及分泌性蛋白质分子--含Ⅲ型纤连蛋白结构域蛋白5(fibronectin type Ⅲ domain containing 5, FNDC5)浓度,测定细胞低氧诱导因子1α(hypoxia-inducible factor-1α, HIF-1α)蛋白的表达水平。结果:(1) 1%低氧浓度组较4%低氧浓度组细胞增殖慢、FNDC5含量低,培养液中葡萄糖浓度高、LDH活性及细胞中HIF-1α蛋白表达较高(P<0.05)。(2)1%低氧浓度条件下,与循环周期6 h组相比,循环周期8 h组细胞活力显著下降(P<0.05)。结论:本研究模拟了体外小鼠成肌细胞在不同低氧/复氧条件下的损伤状况及FNDC5的差异表达,1%低氧浓度和复氧循环周期6 h可作为较佳的体外C2C12细胞实验模型建立条件。
AIM:To establish an in vitro mouse myoblast model of intermittent hypoxia/reoxygenation.METHODS:An O2 content of 1%or 4%in the mixture in a three-gas incubator was established as the hypoxic culture condition,and the environment in a carbon dioxide incubator was established as the reoxygenated culture condition.Mouse myoblasts(C2C12)were exposed to different oxygen concentrations(1%and 4%)and reoxygenation cycles(6 h and 8 h).Then,cell proliferation was detected,and concentrations of glucose,lactate dehydrogenase(LDH)and fibronectin type Ⅲ domain containing 5(FNDC5)in C2C12 cell culture medium were measured.In addition,the expression level of the hypoxia-inducible factor-1α(HIF-1α)protein of C2C12 cells under different experimental conditions was detected by Western blot.RESULTS:(1)The 1%oxygen concentration group showed slower cell proliferation,lower FNDC5 content,and higher glucose levels in culture medium,LDH activity,as well as higher HIF-1αprotein expression compared to the 4%oxygen concentration group(P<0.05).(2)Under 1%oxygen concentration,the 8 h cycle group showed significantly decreased cell viability compared to the 6 h cycle group(P<0.05).CONCLUSION:This study simulates the differences in injury status and FNDC5 expression in mouse myoblasts under different hypoxia/reoxygenation conditions in vitro.A 1%oxygen concentration and reoxygenation cycle of 6 hours can be used as a better experimental model for C2C12 cells in vitro.
作者
席婷
姚晓光
胡君丽
XI Ting;YAO Xiaoguang;HU Junli(Graduate School of Xinjiang Medical University,Urumqi 830001,China;Hypertension Center of Xinjiang Uygur Autono-mous Region People's Hospital,Xinjiang Hypertension Institute,NHC Key Laboratory of Hypertension Clinical Research,Key Laboratory of Xinjiang Uygur Autonomous Region"Hypertension Research Laboratory",Xinjiang Clinical Medical Re-search Center for Hypertension(Cardio-Cerebrovascular)Diseases,Urumqi 830001,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2023年第7期1339-1344,共6页
Chinese Journal of Pathophysiology
基金
新疆维吾尔自治区自然科学基金资助项目(No.2021D01C173)。
