hest value and was in a similar range as the curcuma DMSO e tract. Changes in gene e pression with curcumin Based on the above mentioned findings, curcumin was investigated at different concentrations in more detail at the 6 hour time point. Treat ment with curcumin caused a significant reduction of MMP1 and MMP3 at 10 uM and 20 uM. For MMP13, all concentrations of curcumin caused a significant reduction. E pression of IL 1B and IL 6 was significantly inhibited at both, 10 uM and 20 uM, while the lowest Analysis of NF ��B Immunoblotting of p65 in nuclear e tracts of untreated, IL 1B treated and IL 1B curcumin treated cells revealed that IL 1B treatment caused nuclear translocation of p65 after 60 min. However, compared to IL 1B stimulated samples, curcumin treatment did not reduce levels of the target protein in nuclear e tracts.
Using the NF ��B p65 transcription factor assay, we provide further evidence that IL 1B strongly induced NF ��B DNA bind ing, while cur cumin was not able to reduce levels after IL 1B stimulation. Internal assay controls ensured valid ity of the test. concentration caused a slight increase of IL 6. IL 8 e pression was Batimastat also decreased at 20 uM. In con trast, TNF e pression was significantly increased at all three curcumin concentrations, with the most prominent effects at 20 uM. Furthermore, TNF e pression was also increased upon curcumin treatment alone, while all other target genes remained unaltered under these conditions. TLR2 e pression was sig nificantly reduced with each concentration. For summarized values see Additional file 4 Table S4.
Analysis of MAP kinases Effects of curcumin on MAP kinase activity were investi gated by detection of levels of phosphorylated and unphosporylated p38, ERK and JNK using immunoblotting technique of whole cell e tracts. Results demonstrate that IL 1B treat ment increased levels of phosporylated p38, ERK and JNK after 15 min, which is indicative of activation of these MAP kinases. Treatment with curcumin reduced activity of JNK compared to IL 1B treatment, but further increased levels of p ERK and p p38 compared to IL 1B treatment. Levels of unphosporylated p38, ERK and JNK were similar in all groups. Equal protein loading was confirmed by tubulin detection. Discussion Changes in gene e pression Curcuma is not only an ancient spice, but also a trad itional remedy that has been used in Indian and Chinese medicine to treat indigestion and many other medical issues.
Since the 1970s, the anti inflammatory com pounds called curcuminoids were discovered in the spice, with one being curucmin. Because of its anti inflammatory properties, curcuma and its components have been investigated in osteoarthritis and rheumatoid arthritis during the past one to two decades, while only one paper has been published on the effects of curcumin on intervertebral disc cells so far. Our results clearly show that the different curcuma e tracts influenced cellular behavior in a different manner. While the curcuma