The present study aimed to investigate the correlation between apolipoprotein E (APOE) polymorphisms and the intracellular concentration of Ca2+ in astrocytes in the early stages after an injury. The chondroitin sulfate region of three APOE alleles (epsilon 2, epsilon 3 and epsilon 4) was obtained by reverse transcription-polymerase chain reaction (RT-PCR). A recombinant plasmid, pEGFP-N1-APOE, was constructed and identified by sequencing, while astrocytes were isolated from APOE gene-knockout mice and examined using immunocytochemistry. The recombinant plasmid was transfected into the astrocytes using the liposome-mediated method and cell injury models were constructed by a scratch assay. Laser confocal scanning microscopy (LCSM) was used to detect dynamic alterations in intracellular Ca2+ concentration at 12, 24, 48 and 72 h after injury. Compared with the control group, cells transfected with any of the three alleles demonstrated significant increases in the fluorescence intensity of Ca2+ (P<0.05). The fluorescence intensity of Ca2+ was weak at 12 h after injury, with no statistically significant difference detected between any two groups at this time point (P>0.05). However, the fluorescence intensity increased in a time-dependent manner and at 24, 48 and 72 h post injury, the fluorescence intensity of the epsilon 4 allele-containing cells was significantly higher when compared with that of cells harboring the other two alleles (P<0.05). These results indicate that intracellular Ca2+ overloading may contribute to the deterioration of brain cells and poor outcome subsequent to traumatic brain injury in APOE epsilon 4 carriers.
基金:
Guizhou Science and Technology Department [2013-2332]; National Natural Science Foundation of China [81560227]
第一作者机构:[1]Zunyi Med Coll, Affiliated Hosp, Dept Neurosurg, Zunyi 563000, Guizhou, Peoples R China
通讯作者:
通讯机构:[3]Chongqing Med Univ, Affiliated Hosp 1, Dept Neurosurg, 1 Youyi Rd, Chongqing 400000, Peoples R China[*1]Department of Neurosurgery, The First Affiliated Hospital of Chongqing Medical University, 1 Youyi Road, Yuzhong, Chongqing 400000, P.R. China
推荐引用方式(GB/T 7714):
Wu Haitao,Zhou Shuai,Zhao Hongxin,et al.Effects of apolipoprotein E gene polymorphism on the intracellular Ca2+ concentration of astrocytes in the early stages post injury[J].EXPERIMENTAL AND THERAPEUTIC MEDICINE.2018,15(2):1417-1423.doi:10.3892/etm.2017.5555.
APA:
Wu, Haitao,Zhou, Shuai,Zhao, Hongxin,Wang, Yuyu,Chen, Xiaozhong&Sun, Xiaochuan.(2018).Effects of apolipoprotein E gene polymorphism on the intracellular Ca2+ concentration of astrocytes in the early stages post injury.EXPERIMENTAL AND THERAPEUTIC MEDICINE,15,(2)
MLA:
Wu, Haitao,et al."Effects of apolipoprotein E gene polymorphism on the intracellular Ca2+ concentration of astrocytes in the early stages post injury".EXPERIMENTAL AND THERAPEUTIC MEDICINE 15..2(2018):1417-1423