Specific PFKFB3 Inhibitor Memorably Ameliorates Intervertebral Disc Degeneration via Inhibiting NF-κB and MAPK Signaling Pathway and Reprogramming of Energy Metabolism of Nucleus Pulposus Cells

Figure 2

KAN0438757 (KAN) alleviated tumor necrosis factor-α- (TNF-α-) induced extracellular matrix (ECM) degradation of nucleus pulposus primary (NPP) cells both in vitro and in vivo. (a) PFKFB3 expression levels after negative control, TNF-α, TNF-α with 2.5 μM KAN treatment for 24 hours in NPPs. (b) The gray levels of PFKFB3 were quantified and normalized to β-actin using ImageJ. . (c) Expression of the anabolism and catabolism gene matrix metalloproteins (MMPs) 3, 7, 9, and 13 and aggrecan after negative control, TNF-α, and TNF-α with 2.5 μM KAN treatment for 24 hours in NPPs. . (d) MMP3 and MMP9 expression levels after negative control, TNF-α, and TNF-α with 2.5 μM KAN treatment for 24 hours in NPPs. (e) The gray levels of MMP3 and MMP9 were quantified and normalized to β-actin using ImageJ. . (f) Immunohistochemical staining of MMP3 and MMP 9 expressions. Scale  μm. (g) Alcian blue staining of NP primary cells (P2 generation) on high-density culture after negative control, TNF-α, and TNF-α with 2.5 μM KAN treatment for 7 days. Scale  mm, 400 μm. (h) The integrated optical density (IOD) value was calculated to evaluate the extracellular matrix (ECM) of NP primary cells after the indicated treatment. . Data are expressed as . ns: no significance; ; ; ; .

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