Effect of miR-340-5p targeting at BMP4 on differentiation of neural stem cells in rats

Objective To explore regulation effect of miR-340-5p on regulating bone morphogenetic protein 4(BMP4) expression and the differentiation of rat’s NSCs. Methods NSCs of rats were divided randomly into three groups: normal group (Mock), group with nonsense oligonucleotide (Anti-Con) and group with antisense oligonucleotide of miR-340-5p (Anti-miR-340-5p). The qRT-PCR was used to detect the relative expression of miR-340-5p. The expression of NF200 and MAP-2 was detected by immunocytochemical staining and immunofluorescence, respectively. Western blot was used to detect the expression of BMP4 protein. Results The relative levels of miR-340-5p expression were significantly decreased in Anti-miR-340-5p group (0.14±0.01) compared with that of Mock group(1.01±0.17) and Anti-Con group(1.07±0.13) (P<0.01). Immunocytochemical staining indicated that NF200 was positive in cells of Anti-miR-340-5p group. The proportion of MAP2 positive cells was increased in Anti-miR-340-5p group compared with other groups (P<0.05). Western blot showed the increased expression of BMP4 protein in Anti-miR-340-5p group (0.84±0.09) compared with Mock group(0.53±0.04) and Anti-Con group (0.63±0.09) (P<0.05). Conclusion The miR-340-5p may exert a potential function in regulating differentiation of NSCs into neurons through a negative regulation of BMP4. Key words: miR-340-5p; Bone morphogenetic protein 4; Neural stem cells; Differentiation