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.