鱼腥草注射液诱导人肺腺上皮细胞β防御素2 mRNA表达

Houttuynia Cordata induces expression of human beta-defensin-2 mRNA in pulmonary epithelial cells in vitro

目的:探讨鱼腥草注射液体外刺激人肺腺上皮细胞(SPC-A-1)后人β防御素2(human beta-defensin2,HBD-2)基因表达的情况,并研究HBD-2表达对鱼腥草注射液浓度和作用时间的依赖效应。方法:体外培养SPC-A-1细胞,设置12.5、25、50、100和200μg/ml5个鱼腥草注射液浓度梯度,刺激贴壁生长的SPC-A-1细胞8h,根据细胞HBD-2mRNA表达量明确作用最佳的浓度;最佳作用浓度鱼腥草注射液作用0、1、2、4、8、16、24和48h,根据细胞HBD-2mRNA表达量明确最佳作用时间。SPC-A-1细胞HBD-2mRNA表达的情况应用逆转录聚合酶链式反应技术检测。结果:不同浓度鱼腥草注射液刺激SPC-A-1细胞后,HBD-2表达与刺激剂量(rs=0.829,P=0.042)和刺激时间(rs=0.914,P=0.003)具有一定的相关性,当鱼腥草浓度为100μg/ml、刺激8h时,HBD-2mRNA表达水平达到最高。与空白对照组和白细胞介素1β对照组比较,100μg/ml鱼腥草注射液作用8h可以上调SPC-A-1细胞HBD-2mRNA的表达。结论:鱼腥草注射液可诱导HBD-2基因的表达增强,最佳浓度为100μg/ml,最佳刺激时间为8h。

Objective： To explore the effects of Houttuynia Cordata on expression of human β-defensin-2 （HBD-2） in pulmonary epithelial cells （SPC-A-1） in vitro, and to observe the correlationship between the level of HBD-2 mRNA and the concentrations or treatment times of Houttuynia Cordata. Methods： The SPC-A-1 cells were cultured with different concentrations of Houttuynia Cordata in vitro, including0, 12.5, 25, 50, 100 and 200 μg/ml. And then, the SPC-A-1 cells were cultured with the optimal concentration of Houttuynia Cordata in different lengths of time, including 1, 2, 4, 8,16 and 24 hours. After the treatment, the mRNA level of HBD-2 in pulmonary epithelial cells was detected by means of semiquantitative reverse transcription polymerase chain reaction （RT-PCR）. Results： After being cultured with Houttuynia Cordata, the expression of HBD-2 mRNA had positive correlation with the stimulus concentrations （rs=0.829, P=0.042） and stimulus time （rs=0.914, P= 0. 003）. The highest expression of HBD-2 mRNA was induced by 100 μg/ml Houttuynia Cordata after 8-hour treatment. In comparison with the normal control group and the interleukin-1β group, 100 μg/ml Houttuynia Cordata could significantly up-regulate the expression of HBD-2 mRNA in SPC-A-1 cells after 8-hour treatment （P〈0.01）. Conclusion： Houttuynia Cordata can up-regulate expression of HBD-2 mRNA in SPC-A-1 cells, and the highest expression level of HBD-2 mRNA can be obtained by culture with 100 μg/ml Houttuynia Cordata for 8 hours.