低(无)糖、缺氧和缺氧/复氧对培养鼠脑胶质细胞活力及致损的影响

Effects of glucose,oxygen deprivation,and hypoxia/reoxygenation on the viability and injury of rat astrocytes

目的 :研究氧、糖剥夺和缺氧 /复氧单一或联合因素对培养鼠脑星形胶质细胞活力和引起损伤的影响及其时效反应。方法 :培养的鼠脑星形胶质细胞分为 3组 :无糖组 (D Hanks液 )、低糖组 (DMEM中含 2 .75mmol葡萄糖 )和正常糖组 (DMEM中含 5 .4 9mmol葡萄糖 ) ,各组分别进行不同时间的缺氧和 /或缺氧 /复氧。通以 5 %CO2 +95 %N2 混合气造成缺氧 ,以乳酸脱氢酶 (LDH)漏出率作为细胞受损指标 ,噻唑蓝 (MTT)降解率评测细胞活力。结果 :①在正常氧培养 2 4h ,低糖和无糖培养的细胞LDH漏出率和MTT降解率并不出现明显变化 ;② 3组二项指标在缺氧培养的各时间点依次出现明显变化 ,无糖组为 8h(P <0 .0 5 ) ,低糖组和正常糖组为 12h(P <0 .0 5 ) ,2 4h(P <0 .0 1) ;③缺氧 /复氧引起 3组二项指标出现明显变化 ,其时间点分别为无糖组缺氧 4h +复氧 12h(P <0 .0 5 ) ;低糖组缺氧 8h +复氧 12h (P <0 .0 5 ) ;正常糖组缺氧 8h +复氧 2 4h (P <0 .0 5 ) ;④在同一缺氧和缺氧 /复氧时间点 ,指标值变化程度为无糖组 >低糖组 >正常糖组 ;且两项指标的变化具有时间依赖性 ,呈反变关系。结论 :①氧、糖剥夺和缺氧复氧能引起培养的鼠脑星形胶质细胞的损伤 ;②氧、糖剥夺和缺氧

Objective To explore the single and combined effect of oxygen, glucose deprivation,and hypoxia/reoxygenation on the viability and injury of rat astrocytes and its time relation. Methods Rat cerebral astrocytes were cultured and were divided into 3 groups: normal glucose group, which contained 5.49 mmol glucose DMEM; low glucose group, which contained 2.75 mmol glucose DMEM; and no glucose buffer slat solution group ( D hanks solution). Astrocytes of the 3 groups were exposed to different periods of hypoxia (H) and/or hypoxia/reoxygenation (H/R). The anaerobic mixed gas consisted of 5% CO 2 and 95% N 2. The indexes of cell viability and cell damage or lysis were evaluated by MTT reduction and lactate dehydrogenase (LDH) efflux assay. Results ① Using the no glucose and low glucose culture media, MTT reduction and LDH efflux did not change much during the 24 h normoxia culture. ② Significant changes of the 2 indexes in the 3 groups occurred following the hypoxia time point: no glucose group was at 8 h ( P <0.05),both the low glucose and normal glucose groups were at 12 h ( P <0.05) and 24 h after the hypoxia ( P <0.01);③ Obvious changes of the 2 indexes in H/R were found in the 3 groups at the following time:at 4 h after the hypoxia and 12 h after the reoxygenation ( P <0.05) in the no glucose group, at 8 h after the hypoxia and 12 h after reoxygenation ( P <0.01) in the low glucose group, and at 8 h after the hypoxia and 24 h after the reoxygenation ( P <0.05) in the normal glucose group. ④ The change altitudes of LDH efflux and MTT reduction at the same point after the hypoxia and H/R were in the following order: glucose free group>low glucose group>normal glucose group, which were time dependent. There was a reverse relationship between the changes of LDH and MTT. Conclusion ① Oxygen,glucose deprivation, and H/R can cause rat cerebral astrocyte injury; ② The combined glucose oxygenation deprivation and H/R can increase the cell damage and decrease the cell viability, which are time dependent.