摘要
目的 探讨失血性休克大鼠血清及参附注射液对人脐静脉内皮细胞（HUVEC）内皮细胞蛋白C受体（EPCR）表达的影响.方法 采用酶联免疫吸附试验（ELISA）、逆转录聚合酶链反应（RTPCR）、蛋白免疫印迹法（WB）技术,分别对正常对照组、失血性休克（3,12,24,72 h）组、参附（3,12,24,72 h）组大鼠血清培养的人脐静脉内皮细胞可溶性EPCR（sEPCR）浓度和EPCRmRNA及蛋白表达水平进行检测.结果与正常对照组比较,失血性休克（12,24 h）组、参附（24 h）组的sEPCR及EPCRmRNA含量均显著增高（P＜0.01）,参附（12 h）组的sEPCR及EPCRmRNA含量亦增高（P＜0.05）;失血性休克（12,24 h）组、参附（24 h）组EPCR蛋白含量均显著下降（P＜0.01）,参附（12 h）组EPCR蛋白含量亦下降（P＜0.05）.与失血性休克组（相同时间点）比较,参附（12 h、24 h）组的sEPCR及EPCRmRNA含量明显下降（P＜0.05）,参附（12 h、24 h）组EPCR蛋白含量明显增高（P＜0.05）.结论失血性休克大鼠血清和参附注射液可以从基因、蛋白水平影响人脐静脉内皮细胞EPCR的表达,参附注射液可有效减轻休克内皮细胞损伤.
Objective To investigate the effects of the serum from rats with hemorrhagic shock and Shenfu injection, on the expression of endothelial cell protein C receptor （EPCR） in human umbilical vein endothelial cells （HUVECs） cultured with rat serum. Method The soluble endothelial protein C receptor （sEPCR） in supernatant, the expression of EPCR mRNA and protein level of EPCR in HUVECs were detected by using enzyme linked immunosorbent assay （ ELISA）, reverse transcription polymerase chain reaction （RT-PCR）, and western bloting （WB） in normal control group, hemorrhagic shock serum （3 h, 12 h, 24h, 72 h） group, and Shenfu-treated （3 h, 12 h, 24 h, 72 h） group, respectively. Results The mean levels of sEPCR and the expression of EPCR mRNA were significantly higher in hemorrhagic shock serum （12 h, 24 h） group, and Shenfu -treated（24 h）group than those in normal control group （all P 〈0.01 ）,the mean levels of sEPCR and the expressions of EPCR mRNA were significantly higher in Shenfu-treated （ 12 h） group than those in normal control group （ all P 〈0. 05 ）, while the levels of protein were lower in hemorrhagic shock serum （ 12 h, 24 h） group and in Shenfu-treated（24 h）group than those in normal control group （ both P 〈0.01 ）, and the level of EPCR protein was lower in Shenfu-treated（ 12 h） group than that in normal control group （ P 〈 0. 05） . The mean levels of sEPCR and the expressions of EPCR mRNA were significantly lower in Shenfu-treated （ 12 h, 24 h） group than those in hemorrhagic shock serum （ 12 h,24 h） group （all P 〈0.05）, while the levels of EPCR protein were higher in Shenfu-treated （ 12 h, 24 h）group than those in hemorrhagic shock serum （ 12 h, 24 h） group （ P 〈 0.05 ）. Conclusions These data suggest that Shenfu injectio could affect the expression of EPCR mRNA and the level of EPCR protein, thereby it might be effective in prevention of development of hemorrhagic shock.
出处
《中华急诊医学杂志》
CAS
CSCD
北大核心
2011年第2期168-172,共5页
Chinese Journal of Emergency Medicine
基金
浙江省自然科学基金（Y207635）
