[关键词]
[摘要]
目的 研究丹红注射液对缺氧致原代培养的乳鼠脑微血管内皮细胞(rBMECs)损伤的保护作用。方法 原代培养rBMECs,并对其进行兔抗鼠VIII因子鉴定。实验设对照组,缺氧模型组,丹红注射液低、中、高剂量(25、50、100 μL/mL)组,给药后,rBMECs缺氧4 h。显微镜下观察细胞形态;流式细胞仪检测细胞凋亡率及DNA的量,按试剂盒方法检测细胞上清液中乳酸脱氢酶(LDH)水平、细胞中超氧化物歧化酶(SOD)活性。结果 丹红注射液50、100 μL/mL可显著对抗缺氧造成的损伤,明显改善rBMECs形态,使缺氧所致细胞凋亡数明显减少,有效抑制缺氧诱导的rBMECs发生G1/S期阻滞,抑制LDH释放,增强SOD活性。结论 丹红注射液对缺氧所致原代培养的rBMECs损伤具有明显的保护作用,其机制与增强细胞抗氧化能力、抑制细胞凋亡有关。
[Key word]
[Abstract]
Objective To study the protective effect of Danhong Injection (DI) on primary cultured neonate rat brain microvascular endothelial cells (rBMECs) injury. Methods The primary cultured rBMEC model was established and the identification of rabbit anti rat VIII factor was carried out. The cells were divided into control, model, low-, mid-, and high-dose (25, 50, and 100 μL/mL) DI groups in hypoxic condition for 4 h after administration. The cell morphology was observed under microscope, the apoptosis rate and DNA content were determined by flow cytometry, and the lactate dehydrogenase (LDH) level in cultural supernatants and cell superoxide dismutase (SOD) activity were detected according to the kit methods. Results DI (50 and 100 μL/mL) could alleviate the rBMEC damage induced by hypoxia remarkably, improve the cell morphology of rBMECs, decrease the apoptosis significantly, inhibit the blockage of rBMECs in G1/S phase and the leakage of LDH, and increase the SOD activity. Conclusion DI plays a significant role in the protection on injured primary cultured rBMECs induced by hypoxia, and the mechanism may be related to the enhancement of cellular anti-oxidative capacity and the inhibition of apoptosis.
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[基金项目]
国家自然科学基金资助项目(1173647,81373898,81274176);浙江省自然科学基金资助项目(LR12H27001);浙江省科技厅项目(2013C33244);浙江省中医药(中西医结合)重点学科项目(2012-XK-A06)