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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 effects of Jinlongshe Granule drug-containing serum (JG-DS) on the formation, migration and apoptosis of human lymphatic endothelial cells (HLECs) in vitro. Methods The third generation HLECs were divided into two groups: control group (cultured with saline serum) and experimental group (cultured with JG-DS). After cultured for 12 h in both groups, the ability of tube formation of HLECs was tested by Matrigel tube formation assay; the migration ability of HLECs was detected by Transwell method; the apoptosis rate of HLECs was detected by flow cytometry and Annexin-Ⅴ-FITC / PI double staining. Results The total length of tubules of HLECs after treated with 10% JG-DS for 12 h was (3 084.49 ± 326.27) μm, which was significantly lower than that of the control group (7 058.93 ± 4 567.39) μm, the difference was statistically significant (P <0.01). The migration index of HLECs was (99 ± 26), which was significantly lower than that of the control group (160 ± 32), the difference was statistically significant (P <0.05). The apoptotic rates of HLECs between the two groups showed no significant difference (P> 0.05). Conclusion Jinlongshe granule can inhibit the formation and migration of HLECs in vitro, which may be one of the mechanisms of its inhibition of lymphangiogenesis.