Sulforaphane Delays Intervertebral Disc Degeneration by Alleviating Endoplasmic Reticulum Stress in Nucleus Pulposus Cells via Activating Nrf-2/HO-1

Figure 4

After inhibiting HO-1, the protective effect of SFN on NPC was eliminated. NPCs were cultured under 1% hypoxia to simulate the hypoxia environment of intervertebral disc in vivo. (a, b) The inhibitory effect of si-RNA on HO-1 was detected by WB. ^@@@. (c–f) WB was used to detect the expression of (c, d) ERS-related proteins and (e, f) apoptosis-related proteins, and the relative quantitative data was calculated accordingly. (g) ROS in the NPCs was labeled with CellROX Deep Red Reagent, and the nucleus was dyed blue by DAPI. Representative images were taken by fluorescence microscope. Scale  μm. (h) Quantitative analysis of ROS level. (i, j) TUNEL staining was used to detect the apoptosis of NPCs, and the relative quantitative data was calculated accordingly. Scale  μm. (k, l) The apoptosis of NPCs was detected by flow cytometry. (Error bars showed ; ; , , , vs. control group; ^#, ^##, ^###, vs. AGEs group; ^$, ^$$, ^$$$, vs. AGEs+SFN + si NC group).

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