Transcriptomic Analysis of Vascular Endothelium

In order to explore the mechanism of the protective effect of the combination of Uncaria and Semen Raphani on vascular endothelium, RT-PCR chip for biological function of endothelium was performed to observe mRNA expression of the thoracic aorta [10 (link)]. The total RNA of the thoracic aorta was isolated using the TRIzol method and then reverse-transcribed into cDNA with the PrimeScript RT reagent kit (RR047A, TaKaRa, Dalian, China). Following quantitative detection, the cDNA was hybridized in an Affymetrix Hybridization Oven 640, with subsequent elution inside a clean workstation (GeneChip Fluidics Station 450, Affymetrix). The chips were then scanned with a Gene Array Scanner 3000 7G (Affymetrix) to trace the detection signal. A GeneSpring GX 7.3.1 was used for the gene expression analysis and transcripts with low expression levels (<25% of the median gene expression value) and frequently missed hybridizations (>2 absent flags among the samples) were not analyzed further. Transcripts that showed a >2-fold expression reduction in tissue from drug-treatment groups relative to SHR groups were extracted. The transcript list contained 84 transcripts. Both the test group and control group included three biological replicates [11 (link)].