金龙蛇颗粒含药血清对人淋巴管内皮细胞体外成管、迁移及凋亡的影响

Effect of Jinlongshe Granule Drug-containing Serum on Tube Formation,Migration and Apoptosis of Human Lymphatic Endothelial Cellsin vitro

目的观察金龙蛇颗粒含药血清(Jinlongshe Granule drug-containing serum,JG-DS)对人淋巴管内皮细胞(human lymphatic endothelial cells,HLECs)体外成管、迁移及凋亡的影响。方法制备JG-DS;将第3代HLECs分为2组:对照组(采用生理盐水血清进行培养)和实验组(采用JG-DS培养)。两组细胞培养12 h后,采用Matrigel基质胶成管实验检测HLECs成管能力;Transwell法检测HLECs迁移能力;采用流式细胞术、Annexin-Ⅴ-FITC/PI双染法检测HLECs凋亡率。结果 10%JG-DS作用12 h后,HLECs小管总长度为(3 084.49±326.27)μm,明显低于对照组的(7 058.93±4 567.39)μm,差异有统计学意义(P<0.01);HLECs迁移指数为(99±26)个,明显低于对照组的(160±32)个,差异有统计学意义(P<0.05)。两组HLECs凋亡率比较,差异无统计学意义(P>0.05)。结论金龙蛇颗粒对HLECs体外成管及迁移有抑制作用,可能是其抑制肿瘤微淋巴管生成的机制之一。

Objective To observe the inhibitory effect of Jinlongshe Granule drug-containing serum（ JGDS） on tube formation,migration,and apoptosis of human lymphatic endothelial cells（ HLECs）in vitro. Methods JG-DS was prepared. The 3rd-passage HLECs were divided into the control group（ cultured with normal saline containing serum） and the experimental group（ cultured with JG-DS）. After cultured for 12 h,the tube formation ability was detected by Matrigel assay,and the migration ability was determined by Transwell assay in the two groups. Cell apoptosis rate was detected by flow cytometry and Annexin-Ⅴ-FITC / PI staining method. Results The total length of tube was（ 3 084. 49 ± 326. 27） μm after acted by 10% JG-DS for 12 h,significantly shorter than that of the control group（ 7 058. 93 ± 4 567. 39） μm（P〈0. 01）. The migration number of HLECs was（ 99 ± 26）,obviously lower than that of the control group（ 160 ± 32; P〈0. 05）. The apoptosis rate of the two groups was not statistically significant（P〉0. 05）. Conclusion JG could inhibit the tube formation and migration of HLECsin vitro,which might be one of mechanisms for inhibiting tumor micro-lymphatics.