摘要
目的探讨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）损伤发挥保护作用的可能机制。方法采用胰酶消化法原代培养大鼠RMECs，用Ⅷ因子相关抗原免疫荧光法鉴定RMECs。取生长良好的第3、4代细胞，分为空白对照组、LPS（10mg／L）刺激组和血必净（25mg／ml预处理）治疗组。透射电镜下观察各组细胞形态变化；流式细胞仪检测内皮细胞活性氧（ROS）的表达；免疫荧光法检测核转录因子-κB（NF-κ）活化。结果与空白对照组比较，电镜下LPS刺激组细胞线粒体和粗面内质网空泡变性明显，刺激5min时ROS明显增加[（9．33±0．76）％比（O．03±0．00）％，P〈0．053，刺激30minROS表达至峰值（69．71±4．25）％，LPS刺激30min时NF—κB即完全移位至细胞核内，NF—κB荧光强度在120min内呈平稳上升趋势；与LPS刺激组比较，血必净组细胞形态明显改善，ROS表达量降低[30min：（23．86±2．04）％比（69．71±4．25）％]，NF-KB活化减弱（30min：38．59±2．62比58．71±4．33），差异有统计学意义（均P〈0．05）。结论LPS可刺激细胞产生大量ROS，从而损伤内皮细胞形态，同时通过活化NF-κB影响其功能。血必净注射液可以通过降低ROS表达、减弱NF-κB活化起到保护内皮细胞的作用。
Objective To investigate the protective effects of Xuebijing injection on injury of renal microvascular endothelial cells （RMECs） induced by lipopolysaccharide （LPS） and its possible mechanisms. Methods The rat RMECs were isolated by means of the trypsin digestion technique, and the cultured 3 to 4 well grown passages were identified by FVm-related antigen immunofluorescence. Then they were divided into the blank control group, LPS （ 10 mg/L ） stimulation group and Xuebijing injection （25 mg/ml pretreatment ） treatment group. The transmission electron microscope was applied to observe the changes in cell morphology, flow cytometry, to detect the endothelial cell reactive oxygen species （ROS） expression and immunofluorescence assay, to detect the level of nuclear transcription factor-κB （NF-κB） activation. Results Compared with the blank control group, under the electron microscope, in the LPS stimulation group, the vacuolar degeneration in mitochondria and rough endoplasmic reticulum was significant, ROS were increased obviously [ （9.33 ± 0.76 ）% vs. （0.03 ± 0.00 ）%, P〈0.05 ） in 5 minutes, the expression reached to its peak （69.71 ±4.25 ）% in 30 minutes, and simultaneously, NF-KB completely transferred into the nucleus in LPS stimulation 30 minutes, and the NF-κB fluorescence intensity was in a steady upward trend in 120 minutes. Compared with LPS stimulation group, the cell morphology was improved obviously, ROS expression was decreased [ 30 minutes： （23.86 ± 2.04 ）% vs. （69.71 ± 4.25 ）% 3, NF-κB activation was weakened （30 minutes ： 38.59 ± 2.62 vs. 58.71 ± 4.33 ）in Xuebijing treatment group, the differences being statistically significant （both P〈 0.05 ）. Conclusions LPS can stimulate the RMECs of rat to produce a large amount of ROS, resulting in injury to the cell morphology and affecting its function through the activation of NF-κB. Xuebijing injection can reduce the expression of ROS and weaken the NF-κB activation, thus it can play a protective role for the endothelial cells.
出处
《中国中西医结合急救杂志》
CAS
北大核心
2012年第2期94-97,共4页
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care
基金
基金项目：辽宁省教育厅科研项目（[2008]84）
辽宁省沈阳市科技计划项目（F10-205-1-01）
关键词
血必净注射液
内皮细胞
脂多糖
活性氧
核转录因子
肾脏
中药注射液
Xuebijing injection
Endothelial cell
Lipopolysaccharide
Reactive oxygen species
Nuclear transcription factor
Renal
Chinese traditional medicine injection
