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目的:检测平阳霉素(pingyangmycin PYM)作用于人静脉畸形内皮细胞(Human venous malformation en-dothelial cells HVMECs)后E-selectin表达。方法:通过体外培养HVMECs,采用细胞ELISA和RT-PCR技术检测不同浓度PYM作用HVMECs后E-selectin表达。结果:PYM作用HVMECs后E-selectin在细胞表面表达,在1μg/mL浓度以下具有时间剂量效应。对照组细胞E-selectin表达阴性。PYM作用于HVMECs 6 h后,E-selectin表达达到最高,8 h后表达逐渐下降,24 h后E-selectin在细胞表面无明显表达。1μg/mL PYM作用于HVMECs后6 h,E-selectin表达最高,在10 ng/mL PYM作用于内皮细胞后E-selectin表达明显下降。RT-PCR检测E-selectin mRNA表达,E-selectin在对照组中无明显表达,10 ng/mL以上浓度PYM作用于HVMECs 2 h后E-selectin mRNA表达阳性,表达明显增加。不同浓度PYM作用于HVMECs 6 h后,mRNA表达量最高,12 h后表达逐渐降低,24 h后接近对照组,呈阴性表达。1μg/mLPYM作用于HVMECs后在不同时间段E-selectin表达较高。结论:低浓度PYM能诱导或促进HVMECs E-selectin表达,E-selectin表达与PYM作用时间和药物浓度呈正相关。
Objective: To detect the expression of E-selectin after pingyangmycin PYM treatment on human venous malformation en-dothelial cells (HVMECs). Methods: HVMECs were cultured in vitro. The expression of E-selectin in HVMECs treated with different concentrations of PYM was detected by ELISA and RT-PCR. Results: E-selectin was expressed on the cell surface after HVMECs treated with PYM, with a time-dose effect below 1 μg / mL. The control group cells E-selectin expression was negative. After 6 h of PYM treatment, the expression of E-selectin reached the peak at 6 h, and gradually decreased at 8 h. After 24 h, E-selectin had no obvious expression on the cell surface. The expression of E-selectin was the highest at 6 h after treated with 1 μg / mL PYM, and the expression of E-selectin was significantly decreased after 10 ng / mL PYM treatment. The expression of E-selectin mRNA was detected by RT-PCR. E-selectin was not expressed in the control group. The expression of E-selectin mRNA was significantly increased after treated with PYM at 10 ng / mL for 2 h. After treated with different concentrations of PYM for 6 h, the mRNA level of PYM was the highest at 12 h, and gradually decreased at 12 h. After 24 h, the expression of PYM was nearly negative. E-selectin expression was higher at different time after 1μg / mL PyYM treatment on HVMECs. Conclusion: Low concentrations of PYM can induce or promote E-selectin expression in HVMECs. E-selectin expression is positively correlated with the duration of PYM and drug concentration.