论文部分内容阅读
目的:观察低氧预适应(HPC)对氧糖剥夺(OGD)损伤人神经母细胞瘤细胞(SH-SY5Y)的保护作用,并探讨其可能机制。方法:SH-SY5Y细胞随机分为4组:正常对照组:常规培养,不进行OGD处理;HPC处理组:将神经元放入低氧培养箱内(2%O_2),30 min后立即取出,再恢复常氧培养,反复5次;OGD组:无糖培养基、低氧培养箱内(1%O_2)处理细胞10 h,然后复氧复糖培养24 h;HPC+OGD处理组:细胞HPC后,行OGD处理。通过形态学观察,MTT比色法检测细胞存活率,乳酸脱氢酶(LDH)漏出量判断细胞损伤的程度,原位末端标记(TUNEL)法检测凋亡水平,Western blot检测Caspase 3、低氧诱导因子1α(HIF-1α)的蛋白表达。结果:HPC可减轻OGD引起的SH-SY5Y细胞凋亡,降低LDH漏出量,明显增加OGD组SH-SY5Y细胞的活力(P<0.05)。Western blot显示HPC+OGD组Caspase 3蛋白的表达明显低于OGD组(P<0.05);HIF-1α蛋白的表达明显高于OGD组(P<0.05)。结论:HPC对体外培养的SH-SY5Y细胞OGD损伤具有保护作用,其机制可能与上调HIF-1α蛋白有关。
Objective: To observe the protective effect of hypoxic preconditioning (HPC) on human neuroblastoma cells (SH-SY5Y) induced by oxygen glucose deprivation (OGD) and to explore its possible mechanism. Methods: SH-SY5Y cells were randomly divided into 4 groups: normal control group: normal culture without OGD treatment; HPC-treated group: neurons were placed in hypoxic incubator (2% O 2) OGD group: glucose-free medium, hypoxia incubator (1% O 2) for 10 h, then reoxygenated complex glucose culture 24 h; HPC + OGD treatment group: cell HPC After OGD line processing. Morphological observation, MTT colorimetric assay cell viability, lactate dehydrogenase (LDH) leakage to determine the degree of cell damage, in situ TUNEL assay apoptosis levels, Western blot detection of Caspase 3, hypoxia Inducible factor 1α (HIF-1α) protein expression. Results: HPC could reduce the apoptosis of SH-SY5Y cells induced by OGD, reduce the leakage of LDH, and significantly increase the viability of SH-SY5Y cells in OGD group (P <0.05). Western blot showed that the expression of Caspase 3 protein in HPC + OGD group was significantly lower than that in OGD group (P <0.05). The expression of HIF-1α protein in HPC + OGD group was significantly higher than that in OGD group (P <0.05). CONCLUSION: HPC can protect OGD cells from injury in SH-SY5Y cells in vitro. The mechanism may be related to up-regulation of HIF-1α protein.