Fig. 6

MiR-143-5p suppresses eEF2 and activates the AMPK signaling pathway in NP cells. a miR-143-5p expression and mRNA expression of cyclinD, eEF2, mTOR, Bcl-2, Bax, and AMPK in NP cells from degenerative IVD treated with miR-143-5p mimic, inhibitor, or AICAR determined by RT-qPCR. b, c Protein levels and bands of cyclinD, eEF2, mTOR, Bcl-2, Bax, and AMPK in NP cells treated with miR-143-5p mimic, inhibitor, or AICAR determined by Western blot analysis; AICAR was the activator of the AMPK signaling pathway; the experiment was repeated three times independently; data were expressed by mean ± standard deviation; comparison of data among multiple groups was analyzed using one-way ANOVA; ^*, p < 0.05 vs. the control group (NP cells from normal IVD); ^#, p < 0.05 vs. the blank group (NP cells from degenerative IVD without transfection) and the NC group (NP cells from degenerative IVD transfected with NC sequence); miR-143-5p mimic group refers to NP cells from degenerative IVD transfected with miR-143-5p mimic; miR-143-5p inhibitor group refers to NP cells from degenerative IVD transfected with miR-143-5p inhibitor; AICAR group refers to NP cells from degenerative IVD treated with AMPK signaling pathway activator, AICAR; miR-143-5p inhibitor + AICAR group refers to NP cells of degenerative IVD treated with AICAR and transfected with miR-143-5p inhibitor. NC, negative control; miR-143-5p, microRNA-143-5p; eEF2, eukaryotic translation elongation factor 2; AMPK, AMP activated protein kinase; Bax, Bcl-2 Associated X protein; Bcl-2, B cell lymphoma-2; mTOR, mammalian target of rapamycin; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; NP, nucleus pulposus; ANOVA, analysis of variance