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. 2022 Jun 20;13:905966. doi: 10.3389/fphar.2022.905966

FIGURE 4.

FIGURE 4

Curcumenol mitigated inflammatory reactions in rat’s primary NP cells by inhibiting the NFκB pathway in vitro. (A) Successful isolation of primary NP cells in rats. (B) Western blot analysis of p-P65, P65, p-IκBα, and IκBα expression in primary NP cells pretreated with 50 μM Curcumenol and then stimulated with TNFα (10 ng/ml) for 10 min. (C) Semi-quantification of grey scale value in p-P65/P65 and p-IκBα/IκBα was conducted in panel B. (D) RT-qPCR analysis of the relative mRNA expression levels of Col2a1, MMP3, MMP9, and MMP13 in primary NP cells stimulated with TNFα (10 ng/ml) and 50 μM Curcumenol for 24 h. (E) Western blot analysis of MMP3, MMP9, and MMP13 expression in primary NP cells stimulated with TNFα (10 ng/ml) or/and 50 μM Curcumenol for 24 h. (F) Semi-quantification of grey scale value in MMP3, MMP9, and MMP13 was conducted using β-actin as the reference in panel E. All data are presented as mean ± SD from three replicates. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. Col2a1, collagen type II α 1 chain; IOD, integrated optical density.