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目的:研究平阳霉素(PYM)作用于体外原代的培养人静脉畸形内皮细胞后转化生长因子β1的表达。方法:通过体外原代培养人静脉畸形内皮细胞,采用ELISA和RT-PCR技术检测不同浓度PYM作用人静脉畸形内皮细胞TGF-β1表达。结果:ELISA发现内皮细胞在自然状态下能分泌TGF-β1,体外培养12h后,随着培养时间的增加,TGF-β1的释放量也随之增加。对照组内皮细胞培养48h后,细胞上清中TGF-β1含量达到(537.56±2.61)pg/ml。500ng/ml和1μg/ml PYM作用内皮细胞12h后,内皮细胞表达分泌的TGF-β1较高,且差异有统计学意义。其他浓度PYM作用于静脉畸形内皮细胞24、48h后,TGF-β1表达与对照组细胞差异无统计学意义。PYM浓度≥10μg/ml,内皮细胞TGF-β1表达量较低。RT-PCR结果显示,对照组(未加药物)细胞TGF-β1mRNA阳性表达,PYM浓度为500ng/ml和1μg/ml时TGF-β1mRNA表达较高,PYM浓度在10～100ng/ml之间,TGF-β1mRNA表达量与对照组之间差异无统计学意义。结论:内皮细胞在PYM致静脉畸形组织纤维化过程中能分泌促组织纤维化细胞因子TGF-β1促进组织纤维化。
OBJECTIVE: To study the effect of PYM on the expression of transforming growth factor-β1 in cultured human venous malformation endothelial cells in vitro. Methods: Human primary venous malformation endothelial cells were cultured in vitro and the expression of TGF-β1 in human venous malformation endothelial cells treated with different concentrations of PYM was detected by ELISA and RT-PCR. RESULTS: The endothelial cells secreted TGF-β1 under natural conditions. After 12 hours of in vitro culture, the release of TGF-β1 increased with the increase of culture time. After cultured for 48h, the content of TGF-β1 in the supernatant of the control group reached (537.56 ± 2.61) pg / ml. After treated with 500ng / ml and 1μg / ml PYM for 12h, the expression of TGF-β1 in endothelial cells was higher and the difference was statistically significant. Other concentrations of PYM in venous malformation endothelial cells 24,48h, TGF-β1 expression and the control group, the difference was not statistically significant. PYM concentration ≥10μg / ml, the lower expression of TGF-β1 in endothelial cells. The results of RT-PCR showed that the expression of TGF-β1 mRNA in the control group (untreated) was higher than that in the untreated group. The mRNA expression of TGF-β1 was higher when PYM concentration was 500ng / ml and 1μg / ml, PYM concentration was between 10-100ng / There was no significant difference between the expression of -β1 mRNA and the control group. CONCLUSION: Endothelial cells can secrete promoting tissue fibrosis cytokine TGF-β1 in tissue fibrosis induced by PYM-induced venous malformations and promote tissue fibrosis.