摘要
目的观察血必净注射液对脂多糖（LPS）诱导大鼠肾脏微血管内皮细胞（RMECs）组织因子（TF）表达的影响，并探讨其可能机制。方法采用胰酶消化法原代培养大鼠RMECs，用Ⅶ因子相关抗原免疫荧光法鉴定。取生长良好的3、4代细胞，分为对照组、LPS刺激组和血必净治疗组；采用透射电镜观察各组细胞形态，应用逆转录一聚合酶链反应（RT—PCR）检测细胞内TFmRNA和内皮素（ET）mRNA表达，用免疫荧光法检测核转录因子-κB（NF—κB）活化程度。结果与对照组比较，10mg／LLPS刺激组电镜下可见细胞粗面内质网和高尔基体空泡变性明显；LPS（1～100mg／L）刺激组TF mRNA和ET mRNA表达均随浓度增加显著上调（P〈0．05或P〈0．01），NF—κB核移位明显；血必净（12．5、25．0、50．0g／L）治疗组较LPS刺激组细胞形态明显改善，TFmRNA和ETmRNA表达均明显减少，差异有统计学意义（P〈0．05或P〈0．01），未观察到NF—κB核移位。结论LPS可呈剂量依赖性诱导TFmRNA和ETmRNA表达；血必净注射液可通过降低TFmRNA表达来抑制LPS诱导的微血管内皮细胞凝血系统的启动，其保护作用的机制可能与降低ETmRNA表达并抑制NF—κB活化有关。
Objective To investigate the effect of Xuebijing injection （血必净注射液） on the expression of tissue factor （TF） in renal micro-vascular endothelial cells （RMECs） induced by lipopolysaccharide （LPS） and to explore the possible mechanisms. Methods RMECs were isolated by means of the trypsin digestion technique, and cultured 2 to 4 passages after identification with FⅧ-related antigen immunofluorescence. The healthy cells of the third and fourth passages were collected, and they were divided into three groups： the control group, the LPS stimulating groups and the Xuebijing treated groups in various dosages. Reverse transcription-polymerase chain reaction （RT-PCR） was adopted to measure TF mRNA and endothelin （ET） mRNA expression in the cells. Immunofluorescence analysis was performed to evaluate the activation of nuclear factor-κB （NF-κB）. Transmission electron microscope was applied to observe the morphology of the cells. Results Compared with the control group, obvious degeneration of rough endoplasmic reticulum and golgiosome was observed in the cells that were cultured with 10 mg/L LPS by electron microscope. LPS （1 - 100 mg/L） increased TF mRNA and ET mRNA expression significantly （P〈0. 05 or P〈0.01）. NF-κB apparently translocated from cytoplasm to the nucleus in the LPS stimulated groups. Xuebijing injection （12.5, 25.0, 50.0 g/L） significantly decreased TF mRNA and ET mRNA expression （P〈0.05 or P〈0. 01）, and activation of NF-κB was not observed in the Xuebijing treated groups. Conclusion LPS increases TF mRNA and ET mRNA expression in a dose-dependent manner. Xuebijing injection inhibits the initial process of coagulation by decreasing the LPS-indueed TF mRNA expression in the micro-vascular endothelial cells. The possible mechanisms may be related to the reduction of ET mRNA expression and the inhibition of the activation of NF-κB.
出处
《中国中西医结合急救杂志》
CAS
北大核心
2009年第4期218-222,F0003,共6页
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care
基金
辽宁省教育厅基金项目（2008836）
