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0.05). The expression of GFP was located around the ventricle and persisted for a long time. Conclusion The fluorescence of GFP is stable and persistent and the GFP has no negative effects on the NSCs-GFP. It is an ideal maker of neural stem cells and could be used for the study of NSCs transplantation.
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Objective To observe DAT1 expression in neural stem cells (NSCs) and the effects of DAT1 on the differentiation of NSCs. Methods Eukaryotic expression vectors pEGFP-C1-DAT1 and pcDNA4/hisA-DAT1 were constructed and transfected into NSCs of rat cerebellum by lipofectamine2000. The transfected NSCs were observed by immunohistochemistry under fluorescent microscope. Results The overexpression of DAT1 could increase the number of Mash-1 staining cells and promote the NSCs to differentiate into neuron progenitors, and the high levels of DAT1 in NSCs facilitated the differentiation of neurons. The localizations of DAT1 protein in Mash-1 staining cells and NF 200 staining cells changed. This shift may result from the two distinct inducing factors, FBS and nature differentiation, or distinct stages in differentiation of NSCs. Conclusion DAT1 functions in differentiation of NSCs as a multiprotein combined with distinct transcription factors by virtue of different inducer or varied stage of differentiation.
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AIM: The aim of this study was to determine the relationship between PAI-1,TIMP-1 gene expression and renal tubulointerstitial fibrosis(TIF) of ureteral obstruction ,and the interfering effects of hepatocyte growth factor (HGF) treatment. METHODS: Sixty rats were divided into normal control, sham operation,unilateral ureteral obstruction(UUO),and rhHGF treated groups (received 0 5 mg?kg -1 ?d -1 HGF for twenty-one days), and were sacrificed at postoperative day 3, 7, 14, 21. The levels of PAI-1, TIMP-1 mRNA were measured by RT-PCR. MMP2 and MMP9 activities were detected by substrate zymography, and renal fibrosis was assessed by measuring tissue hydroxyproline. RESULTS: Compared to controls, expression levels of PAI-1,TIMP-1 mRNA were significantly increased in UUO rats, and this was accompanied by decreased activities of MMP2 and MMP9 and increase in tissue hydroxyproline content. HGF treatment significantly decreased expressions of PAI-1, TIMP-1 mRNA, increased MMP2 and MMP9 activities,and decreased tissue hydroxyproline content in the obstructive kidney. CONCLUSIONS: These results indicate that the increases in PAI-1, TIMP-1 mRNA expression may be the major cause of sustained decreased matrix degradation during the development of tubulointerstitial fibrosis after unilateral ureteral obstruction. rhHGF efficiently ameliorates renal tubulointerstitial injury by the reduction of PAI-1,TIMP-1 mRNA expression, and increasing MMP2, MMP9 activities. [