Bushen Huoxue Formula Modulates Autophagic Flux and Inhibits Apoptosis to Protect Nucleus Pulposus Cells by Restoring the AMPK/SIRT1 Pathway

Figure 3

BSHXF treatment antagonizes cell apoptosis, decreases ROS production, and alleviates mitochondrial dysfunction induced by TNF-α. (a–b) Typical images and expression of ROS levels. The phalloidin (green) was used to counterstain the cells. Scale bar: 100 μm. (c–e) Typical fluorescence images of NP cells mitochondria. Phalloidin (green) was used to counterstain cells. Scale bar: 100 μm. (f–g) The determination of the ATP and SOD levels. (h–k) Western blotting was applied to detect the expression of c-caspase-3, mito-cyt, and cyto-cyt.(l and m) The NP cell image of TUNEL staining in each group. The DAPI was used to stain nuclei. Scale bar: 100 μm. (n) Measurement of NP cell apoptosis by flow cytometry. The group treated with serum containing BSHXF, and ; the TNF-α group, ^# and ^##.

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)

(i)

(j)

(k)

(l)

(m)

(n)