GBLP在人脐静脉内皮细胞中差异表达的验证

Verification of differential expression of GBLP in HUVEC treated by R/S-propranolol

目的验证经手性药物比较蛋白质组学筛选鉴定的差异表达蛋白G蛋白β亚单位2样1蛋白(guanine nu-cleotide-binding protein subunit beta2-like1,GBLP)。方法分别用80μmol/L的R/S-普萘洛尔(R/S-propranolol,R/S-PRO)处理人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)3h。采用比较蛋白质组学方法分离鉴定差异表达蛋白-GBLP,Western blot法和激光共聚焦扫描显微镜(LSCM)分别验证经R/S-PRO作用后,HUVEC细胞中GBLP的表达丰度差异和细胞内定位。结果Western blot和LSCM都证实,GBLP在S-PRO作用后的HUVEC中的表达丰度明显低于经R-PRO作用后的表达(P=0.011);GBLP定位于HUVEC的细胞膜和细胞质中,在细胞质中呈均匀分布,在膜上有局部聚集分布现象。结论Western blot和LSCM验证结果与蛋白质组学鉴定结果一致。

Objective To verify the differential expression of the protein - guanine nucleotide-binding protein subunit beta 2-like 1 （GBLP） that has been identified by comparative proteomics methods. Methods Human umbilical vein endothelial cells （HUVEC） were treated with R/S-propranolol （R/S-PRO） respectively for 3 h, then the differentially expressed GBLP protein was isolated and identified by comparative proteomics methods. Western blot analysis and the confocal laser scanning microscopy （LSCM） were used to further verify the expression abundance and display location of GBLP in two enantiomers treated HUVEC. Results Both resuits of Western blot and LSCM exhibited that the expression abundance of GBLP in S-PRO treated HUVEC was lower than that in R-PRO treated cells. GBLP located in cytoplasm and plasma membrane in HUVEC, homogeneously distributed in cytoplasm and aggregatively distributed in plasma membrane. Conclusion The differential expression of GBLP proven by Western blot and LSCM is consistent with that identified by proteomics methods.