Effects of B lymphocytes Invitro Treatment with Rose Extract on Transmembrane Ligands

Abstract

AimsThe study was performed to evaluate the role of red rose extract (Pierre de Ronsard) on B lymphocytes. Specifically the gene expression of CD20, CD30, CD40, and CCR5 in human B cells were studied after treatment with rose extract.MethodsRed rose extract was prepared at the dilution of 0.0075% (v/v) and stored until use at -20°C. Cell treatment was performed at 37°C on B cells. The cells were plated in 6 well plates at 1.5×106cells per well and stored at -80°C. Total RNA extraction and quality control were performed. RTq-PCR was performed according to Genecopoeia’s instructions. The cycle threshold method (ΔΔCt) was used for data analysis.ResultsThe comparative Ct method quantification (2^-ΔCt) and fold change for CD20, CD 30, CD 40 and CCR5 were - 5.65E+01, 4.80E-01, N/A, 2.47E-01; and 0.954,0.377, N/A and 0.577, respectively. The amount of total RNA extracted from about 4.5×106cells was low and did not allow us to measure the RNA profile. The A260/A230 ratios were very low due to the low amount of RNA. The analysis of gene expression by qRT-PCR showed that CD40 was not expressed in untreated cells and cells treated with rose extract with the Ct values over 32. All other genes were expressed and well-measured in both B cell samples.ConclusionThe treatment with rose extract at 0.0075% (v/v) did not modify the expression of CD20. However, the expression of CD30 and CCR5 decreased with the rose extract treatment. The range of the fold change showed that the result of CD30 expression was more accurate than for CCR5.