as constricted at concentrations of 500 ug ml and 1000 ug ml. A Ponatinib dna slight proliferative effect was observed for 100 ug ml and 250 ug ml. For the curcuma ethanol e tract, no cytoto ic effect could be observed at any time point up to a concentration of 1000 ug ml. For curcumin, cytoto ic effects could be observed at concentrations of 50 uM and 100 uM. Changes in gene e pression with IL 1B prestimulation With IL 1B treatment, we could observe a significant in crease in the mRNA levels of all genes of interest at the time of analysis. Data for all genes is shown in Table 3 as mean, SEM and p value. Changes in gene e pression with curcuma DMSO and ethanol e tracts As shown in the Supplementary Material, neither DMSO nor EtOH at the used concentration influenced the e pression of the inflammatory and catabolic target genes.
Treatment with the curcuma DMSO e tract resulted in a significant inhibition of MMP1, MMP3 and MMP13 after 6 hours, relative to IL 1B prestimulated cells. While no changes occurred in the e pression of IL 1B and IL 8, a significant inhibition of IL 6 was observed. However, we noticed a strong induction of TNF e pression at this early time point. E pression of TLR2 was significantly reduced. For all results see Figure 2a as well as Additional file 3 Table S3 for sum marized values. Compared to IL 1B prestimulated cells, treatment with the curcuma EtOH e tract did not cause any changes in gene e pression after 6 hours for MMP1 and MMP3 while slightly decreasing MMP13 e pression. E pression of IL 1B, IL 6 and IL 8 also remained unchanged, but TNF e pression was increased.
TLR2 e pression was not influenced. For all results see Figure 2b as well as Additional file 3 Table S3 for summarized values. Analysis of the curcuma DMSO and EtOH e tracts Based on the above shown results, the DMSO fraction seemed to contain one or more anti catabolic Drug_discovery and anti inflammatory substances. Taking the solubility of the various components of curcuma as well as the literature based preselection of anti inflammatory components of curcuma into account, the curcuminoid curcumin was chosen to be the most promising candidate substance with biological activity. In order to proof that curcumin was indeed present in the DMSO e tract, HPLC MS ana lysis was performed on the stock e tracts. The results showed that predominantly curcumin was present in the e tract at m z 369.
1 followed by its precursors demetho ycurcumin at m z 339. 1 and bisdemetho ycurcumin at m z 309. 1 and other unidentified compounds with little absorbance. As curcumin is also soluble in EtOH, we performed a sequential e traction process described under Materials ARQ197 purchase and Methods in order to aggregate curcumin in the DMSO e tract. Both, the sequential EtOH e tract as well as the pure curcu min stock solution in DMSO were also mea sured by HPLC MS. While the curcuma DMSO e tract contained 6. 32 mg ml of curcumin, the sequential curcuma EtOH e tract contained only 1. 2 mg ml. Curcumin itself showed the hig