There are several major pathological changes in Alzheimer＇s disease, including apoptosis of cho- linergic neurons, overactivity or overexpression of 13-site amyloid precursor protein cleaving enzyme 1 （BACE1） and i...There are several major pathological changes in Alzheimer＇s disease, including apoptosis of cho- linergic neurons, overactivity or overexpression of 13-site amyloid precursor protein cleaving enzyme 1 （BACE1） and inflammation. In this study, we synthesized a 19-nt oligonucleotide targeting BACE1, the key enzyme in amyloid beta protein （AI3） production, and introduced it into the pSilenCircle vector to construct a short hairpin （shRNA） expression plasmid against the BACE1 gene. We transfected this vector into C17.2 neural stem cells and primary neural stem cells, resulting in downregulation of the BACE1 gene, which in turn induced a considerable reduction in reducing AI3 protein production. We anticipate that this technique combining cell transplantation and gene ther- apy will open up novel therapeutic avenues for Alzheimer＇s disease, particularly because it can be used to simultaneously target several pathogenetic changes in the disease.展开更多
Histone deacetylation is a key modulator involved in cell proliferation,apoptosis,and mRNA transcription.However,the effects of histone deacetylation on C17.2 neural stem cells(NSCs)remain unclear.Here,the histone dea...Histone deacetylation is a key modulator involved in cell proliferation,apoptosis,and mRNA transcription.However,the effects of histone deacetylation on C17.2 neural stem cells(NSCs)remain unclear.Here,the histone deacetylase inhibitors nicotinamide and trichostatin A(TSA)were used to determine the role of histone deacetylation on gene transcription in NSCs.The results showed that the mRNA expression of p53,Sox1,Sox2,and Bax were significantly higher in E14.5 NSCs than in C17.2 NSCs.Nestin,a marker gene of neuronal differentiation,did not differ significantly between E14.5 NSCs and C17.2 NSCs.The transcription levels of p53 and Nestin were significantly higher in C17.2 NSCs than in differentiated brain tissues,and the expression of Bax,Sox1,and Sox2 was higher in the olfactory bulb than in other brain tissues.Nicotinamide and TSA treatment decreased the transcription of Sox2,p53,Nestin,and Bax in C17.2 NSCs,although the difference was statistically significant only for Sox2 and Nestin,Sox1 transcription was not detected.These results demonstrated that mRNA expression profiles differ between C17.2 NSCs,E14.5 NSCs,and adult mouse brain tissues,and HDAC inhibitors regulate gene expression by modulating histone acetylation.展开更多
基金supported by grants from the National Natural Science Foundation of China,No.81271476the Natural Science Foundation of Guangdong Province,No.S2011010004366
文摘There are several major pathological changes in Alzheimer＇s disease, including apoptosis of cho- linergic neurons, overactivity or overexpression of 13-site amyloid precursor protein cleaving enzyme 1 （BACE1） and inflammation. In this study, we synthesized a 19-nt oligonucleotide targeting BACE1, the key enzyme in amyloid beta protein （AI3） production, and introduced it into the pSilenCircle vector to construct a short hairpin （shRNA） expression plasmid against the BACE1 gene. We transfected this vector into C17.2 neural stem cells and primary neural stem cells, resulting in downregulation of the BACE1 gene, which in turn induced a considerable reduction in reducing AI3 protein production. We anticipate that this technique combining cell transplantation and gene ther- apy will open up novel therapeutic avenues for Alzheimer＇s disease, particularly because it can be used to simultaneously target several pathogenetic changes in the disease.
基金supported by the Key Scientific Research Projects of Higher Education Institutions in Henan Province under grant number 19A180021the Young Key Teachers Training Program of Yellow River Conservancy Technical Institute+2 种基金the Campus Scientific and Research Fund Project of Yellow River Conservancy Technical Institute under grant number 2017QNKY012the Research Projects of Employment and Entrepreneurship of Secondary and Higher Education Institutions in Henan Province under grant number JYB2018534National Natural Sciences Foundation of China under grant number 31070954.
文摘Histone deacetylation is a key modulator involved in cell proliferation,apoptosis,and mRNA transcription.However,the effects of histone deacetylation on C17.2 neural stem cells(NSCs)remain unclear.Here,the histone deacetylase inhibitors nicotinamide and trichostatin A(TSA)were used to determine the role of histone deacetylation on gene transcription in NSCs.The results showed that the mRNA expression of p53,Sox1,Sox2,and Bax were significantly higher in E14.5 NSCs than in C17.2 NSCs.Nestin,a marker gene of neuronal differentiation,did not differ significantly between E14.5 NSCs and C17.2 NSCs.The transcription levels of p53 and Nestin were significantly higher in C17.2 NSCs than in differentiated brain tissues,and the expression of Bax,Sox1,and Sox2 was higher in the olfactory bulb than in other brain tissues.Nicotinamide and TSA treatment decreased the transcription of Sox2,p53,Nestin,and Bax in C17.2 NSCs,although the difference was statistically significant only for Sox2 and Nestin,Sox1 transcription was not detected.These results demonstrated that mRNA expression profiles differ between C17.2 NSCs,E14.5 NSCs,and adult mouse brain tissues,and HDAC inhibitors regulate gene expression by modulating histone acetylation.
