Antiaging Factor Klotho Retards the Progress of Intervertebral Disc Degeneration through the Toll-Like Receptor 4-NF-κB Pathway

Figure 2

Decreased Klotho expression in NP cells treated with H₂O₂. (a) Primary NP cells were stained by SOX9, collagen II, and Aggrecan. To confirm the purity of in vitro primary NP cells culture, one representative experiment of six was shown. Scale bar, 20 μm (original magnification, ×40). (b) Klotho protein expression was tested from human renal tubule epithelium HK2 cells, mouse macrophage RAW264.7 (RAW), and NP cells by Western blotting. (c) NP cells were treated with increasing amounts of H₂O₂ (0, 50, 100, 200, 400, and 800 μM) for 12 h, and then Klotho and TLR4 protein levels were assayed by Western blotting. (d) NP cells were treated with increasing amounts of H₂O₂ (0, 50, 100, 200, 400, and 800 μM) for 6 h, and then IL-1β, NOS2, and IL-18 mRNAs were assayed by qRT-PCR. GAPDH served as an internal control. The results were presented as of three independently performed experiments. , vs. control.

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