Effect of Rabbit Epididymal Antimicrobial Peptide, REHbP, on LPS-Induced Proinflammatory Cytokine Responses in Human Vaginal Cells In Vitro
Figure 6
RT-PCR analysis of hBD-1 gene expression in hVECs. Cells were seeded at a density of 106/well in a 24-well plates and treated with LPS (10 g/mL for 6 h) or LPS-induced (10 g/mL for 6 h) cells were treated with REHbP (60.61 M for 1 h) or scrambled peptide (60.61 M for 1 h). At the end of treatment, cells were collected and lysates were prepared and analyzed for hBD1 and GAPDH mRNAs by RT-PCR as detailed in Section 2. (a) Expression of hBD1 mRNA was upregulated in LPS-induced cells and attenuated following the treatment of LPS-induced cells with REHbP with respect to the scrambled peptide. Values are calculated as the mean ± SD of triplicate determinations. Representative image of RT-PCR analysis of hBD-1 mRNAs expression is shown. GAPDH blots confirmed roughly equivalent loading of RNA samples. (b) A quantitative assessment of the intensity of the each band was determined by densitometry. Levels of significance ( compared with LPS-induced groups, compared with medium control) were calculated by ANOVA test followed by a Bonferroni analysis.
