清开灵对小鼠脑微血管内皮细胞缺氧损伤MLCK基因转录的影响

Effect of Qingkailing on Transcription of MLCK Gene of Brain Microvascular Endothelial Cells Caused by Hypoxia

目的:研究清开灵对小鼠脑微血管内皮细胞缺氧损伤MLCK基因转录的影响。方法:细胞分为正常对照组(有糖Earle液)、模型组(缺氧缺糖模拟缺血:无糖Earle液,37℃,5%CO2,1%O2培养24 h)、清开灵低、中、高浓度组(缺氧缺塘+清开灵1%、2%、4%),在37℃、5%CO2培养箱中培养24 h后经MTT比色法检测细胞活力、荧光定时定量PCR检测MLCK基因转录。结果:缺氧缺糖24 h后微血管内皮细胞活性降低(P<0.05)、MLCK基因转录被抑制(P<0.05);清开灵1%、2%、4%组均抑制了缺糖缺氧条件下微血管内皮细胞活性的降低(P<0.05),清开灵4%组降低MLCK的表达(P<0.05)。结论:缺糖缺氧条件下微血管内皮细胞活性降低。清开灵通过抑制MLCK的表达,恢复BBB的部分屏障功能。

Objective： To study the effect of Qingkailing on the damaged expression of MLCK gene transcription of brain microvascular endothelial cells caused by hypoxia injury. Methods： The cells were divided to normal control group（ earle medium with sugar）,model group（ earle medium without sugar,37 ℃,5% CO2,1% O2 cultured 24 h）,Qingkailing low,medium and high concentrations groups（ hypoxia and lack of pond ＋ Qingkailing 1%,2%,4%）,incubated for 24 h in5% CO2 incubator at 37 ℃,and then we detected cell vitality by MTT colorimetric method and tested MLCK gene transcription by PCR. Results： Due to hypoxia,the activity of microvascular endothelial cells was reduced（ P〈0. 05） and the expression of MLCK gene transcription was suppressed（ P〈0. 05）. The groups Qingkailing of 1%,2%,4% inhibited the decrease of the activity of microvascular endothelial cells due to hypoxia（ P〈0. 05） and Qingkailing of 1% inhibited the MLCK gene transcription. Conclusion： The activity of microvascular endothelial cells can be reduced due to hypoxia. Qingkailing inhibits the decrease of the activity of microvascular endothelial cells and inhibit the MLCK gene transcription and partly recovers the barrier function of BBB.