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

Figure 4

Mangiferin alleviated the pathological reactions associated with IVDD caused by TNF-α in cultured mouse IVD tissues. The IVD tissues from mice were treated with PBS () or TNF-α (20 ng/ml, ) in the presence or absence of mangiferin (100 μM/ml, ; 500 μM/ml, ). (a–e) Total protein was extracted, and levels of the COX-2, iNOS, ADAMTS-5, and MMP-13 proteins were evaluated using Western blotting. (f) Safranin O staining was performed after treating the mouse IVD tissues with TNF-α and different concentrations of mangiferin. Scale bar: 100 μm. (g, i) The expression level of aggrecan in mouse IVD tissues was detected using IHC staining. Scale bar: 100 μm. (h, j) IHC staining showing the level of Col-2 in mouse IVD tissues. Scale bar: 100 μm. (k–n) We detected the expression levels of COX-2 and MMP-13 using IF staining. Scale bar: 100 μm. (o–r) We detected the expression levels of Bax, Bcl-2, and c-caspase-3 using Western blotting. (s, t) TUNEL staining of the IVD tissues from mice in each indicated group. Scale bar: 100 μm. All the experiments were repeated at least three times. Significant differences are indicated as follows: , , , and .

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