Bridging effect of nuclear factor kappa B on upregulation of vascular endothelial growth factor expression in hypoxia-inducible factor-1 alpha gene-modified neural stem cells
10.3969/j.issn.1673-8225.2009.49.012
- VernacularTitle:核因子kB在低氧诱导因子1α基因修饰神经干细胞上调血管内皮生长因子表达中的桥接作用
- Author:
Yuling JIN
;
Jingjing LIU
;
Xiaofeng ZHU
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2009;13(49):9668-9672
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Numerous studies have shown that nuclear factor-KB (NF-kB) was positively correlated with vascular endothelial growth factor (VEGF) in the cerebral hypoxia region. Thus, we assumed whether NF-kB in the pathway of hypoxia-inducible factor-1 α (HIF-1α) upregulating VEGF and plays a bridge effect.OBJECTIVE: To study the effects of NF-kB in HIF-1α pathway of increasing expression of VEGF using HIF-1α-modified neural stem cells (NSCs) as vectors.DESIGN, TIME AND SETTING: In vitro observation of cytology was conducted at the Neuroscience Institute, Jiamusi University from March to December 2008.MATERIALS: Wistar rats aged less than 24 hours of both genders were used.METHODS: NSCs were transfected after amplification of adenovirus vector HIF-1α (AdHIF-ia)-green fluorescent protein (GFP).Fluorescence detection was used to determine HIF-1α-GFP and blank vector Ad-GFP expression in NSCs. Protein was extracted from transfected NSCs, blank vector NSCs and normal NSCs, separately. Subsequently, NF-kB specific inhibitor pyrrolidine dithiocarbamate (PDTC) (50,150, 300 μmol/L) was added in HIF-1α-modified NSCs. Western blot analysis was used to determine changes in VEGF expression.MAIN OUTCOME MEASURES: The following parameters were measured: expression of HIF-1α, VEGF and NF-kB in NSCs in each group; VEGF expression in NSCs following treatment of NF-kB specific inhibitor.RESULTS: Gene expression was associated with MOI and transfected time following AdHIF-1α-GFP transfected with NSCs. After transfected AdHIF-1α-GFP in NSCs, HIF-1α, VEGF and NF-kB expression was positively correlated. Expression of VEGF was reduced in AdHIF-1α-GFP-modified NSCs following treatment of NF-kB inhibitor PDTC in a concentration-dependent fashion.There were significant differences in VEGF expression between each concentration group (P < 0.05-0.01).CONCLUSION: NF-kB in signaling pathway of HIF-1α-upregulated VEGF expression and played a bridging effect.
