Experimental protocol were performed by instruction manual of lightcycler-rna amplification kit sybr green i . human named genes hgec-40s were used for differential expression screening 以未作任何處理的內(nèi)皮細胞作為對照。內(nèi)皮細胞切應力加載完成后,提取細胞總rna。
Objective to investigate the effects of fluid shear stress on il-8 gene in human umbilical vein endothelial cells ( huvecs ) and the roles of time course of low shear stress and intensity of fluid shear stress using lightcycler ? system; to investigate the gene expression profiles in huvecs exposed on low shear stress ( 4.20 dyne / cm2, 2 h ) and incubated by 17 p-estradiol ( 10-7 m ) + low shear stress ( 4.20 dyne / cm2, 2 h ) using the cdna microarray approach . methods endothelial cells were isolated from human umbilical cord veins by collagenase treatment as described by jaffe and modified 目的探討人臍靜脈內(nèi)皮細胞(humanumbilicalveinendothelialcells,huvecs)在流體切應力作用下il-8基因的誘導表達以及切應力的作用時間和作用強度對il-8基因表達影響的變化規(guī)律;利用表達譜基因芯片技術研究內(nèi)皮細胞在低切應力(4.20dynecm~2,2h)作用下其切應力相關基因的表達情況以及生理濃度(10~(-7)的17-雌二醇孵育內(nèi)皮細胞48h,再經(jīng)同樣條件的切應力處理后對內(nèi)皮細胞切應力相關基因表達的可能影響。