Mangiferin Alleviates Mitochondrial ROS in Nucleus Pulposus Cells and Protects against Intervertebral Disc Degeneration via Suppression of NF-κB Signaling Pathway

Figure 3

Mangiferin alleviated TNF-α-induced mitochondrial dysfunction, oxidative stress, and apoptosis in HNPCs. HNPCs were treated with TNF-α (20 ng/ml) and different concentrations of mangiferin (100 μM/ml and 500 μM/ml). (a) The mitochondrial morphology in HNPCs was observed using TEM. Scale bars: 5 μm and 1 μm. (b–e) The levels of OPA1, TFAM, and Drp1 were assayed using Western blotting. (f, g) JC-1 was used to detect the mitochondrial membrane potential in HNPCs. Scale bar: 100 μm. (h, i) ROS levels were detected in HNPCs from each treatment group. Scale bar: 100 μm. (j, k) MitoTracker was used to detect the mitochondrial membrane potential of HNPCs. Scale bar: 25 μm. The expression of (l) Bax, (m) Bcl-2, and (n) c-caspase-3 in each indicated group was assayed using real-time PCR. (o–r) The levels of Bax, Bcl-2, and c-caspase-3 were assayed using Western blotting. (s, u, v) Hoechst 33342 and (t, w) TUNEL staining of the HNPCs in each indicated group. Scale bar: 100 μm. All the experiments were repeated at least three times. Significant differences are indicated as follows: ns , , , , and .

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)

(i)

(j)

(k)

(l)

(m)

(n)

(o)

(p)

(q)

(r)

(s)

(t)

(u)

(v)

(w)