青蒿琥酯对急性单核细胞白血病SHI-1细胞株血管内皮生长因子及其受体表达的影响

目的 研究青蒿琥酯对急性单核细胞白血病SHI-1细胞株血管内皮生长因子(VEGF)及其受体( VEGFR)的影响。方法酶联免疫吸附法检测非细胞毒性浓度(5、10、20 ng/ml)青蒿琥酯作用SHI-1细胞后培养上清液VEGF浓度，流式细胞术检测有或无青蒿琥酯作用时，SHI-1细胞表面VEGFR-1及VEGFR-2阳性表达率。结果培养24、48 h后，无青蒿琥酯作用的SHI-1细胞培养上清液VEGF质量浓度分别为( 980.3±2.2)、(982.4±2.3) pg/ml，VEGFR-1表达率分别为(5.40±3.11)％和(4.45±2.85)％，VEGFR-2表达率分别为(13.90.± 2.26)％和（13.95±1.96）％。5、10、20 ng/ml青蒿琥酯作用24h后，SHI-1细胞培养上清液VEGF质量浓度分别为(234.6±1.8)、(114.9±1.6)、(108.8±1.5) pg/ml，作用48 h后分别为(62.3±1.7)、(60.9±1.6)、(32.7±1.7) pg/ml，与培养相同时间无青蒿琥酯组相比，VEGF浓度明显下降（均P＜ 0.05），且相同浓度青蒿琥酯作用24 h与48 h间差异亦有统计学意义(均P＜ 0.05)。5、10、20 ng/ml青蒿琥酯作用24 h，VEGFR-1阳性率分别为(4.30±2.21)％、(4.20±1.37)％和(3.90±1.86)％，作用48 h后分别为(3.80±2.87)％、(3.60±1.73)％和(3.00±1.82)％，相同作用时间不同浓度青蒿琥酯组间及相同浓度作用不同时间组间VEGFR-1阳性率差异均无统计学意义(均P＞ 0.05)；作用24h后，SHI-1细胞VEGFR-2阳性率分别为(4.40±1.15)％、(3.10±0.68)％和(1.10±0.72)％，作用48 h后分别为(3.00±1.68)％、(2.20±0.93)％和(0.60±0.92)％，3个不同浓度青蒿琥酯作用相同时间后VEGFR-2表达率降低（均P＜ 0.05），相同浓度作用24与48 h间差异均无统计学意义（均P＞ 0.05）。结论SHI-1细胞株高分泌VEGF，青蒿琥酯可下调VEGF分泌及VEGFR-2的表达，而对VEGFR-1表达的调节作用不显著。

Effect of artesunat on the expression of vascular endothelial growth factor and vascular endothelial growth factor receptors in SHI-1 cell line

Objective To investigate the effect of artesunate on the expression of vascular endothlial growth factors (VEGF) and VEGFR in SHI-1 cell line. Methods Enzyme-linked immunosorbent assay analysis was performed to detect the amount of VEGF in culture supernatants of SHI-1 cell in the condition of artesunate or not. The expression of VEGFR1 and VEGFR2 in SHI-1 cell in the condition of artesunate or not were detected by flow cytometry. Results Without artesunate, the concentration of VEGF in the culture supernatant of SHI-1 cell were (980.3±2.2) pg/ml in 24 h and (982.4±2.3) pg/ml in 48 h. The expression of VEGFRI in SHI-1 cell were (6.40±3.11) ％ in 24 h and (6.45±2.85) ％ in 48 h. The expression of VEGFR2 in SHI-1 cell were (13.90±2.26) ％ in 24 h and (13.95±1.96) ％ in 48 h. With artesunate at 5, 10, 20 ng/ml, the concentration of VEGF in culture supematant of SHI-1 cell were (234.6±1.8) pg/ml, (114.9±1.6) pg/ml, (108.8±1.5) pg/ml in 24 h and (62.3±1.7) pg/ml, (60.9±1.6) pg/ml, (32.7±1.7) pg/ml in 48 h, respectively. The levels of VEGF in SHI-1 cells treated with artesunate at different concentrations decreased significantly (P ＜0.05). There was significant difference between 24 hours group and 48 hours group (P ＜0.05). The expression of VEGFR1 in SHI-1 cell were (4.30±2.21) ％, (4.20±1.37) ％, (3.90±1.86) ％ in 24 h and (3.80±2.87) ％, (3.60±1.73) ％, (3.00±1.82) ％ in 48 h, respectively. The expression of VEGFR1 in SHI-1 cell treated with artesunate at different concentrations were not significantly different (P ＞0.05). No significant difference between 24 hours group and 48 hours group was observed (P ＞0.05). VEGFR2 expression of SHI-1 cell were (4.40±1.15) ％, (3.10±0.68) ％, (1.10±0.72) ％ in 24 h and (3.00±1.68) ％, (2.20±0.93) ％, (0.60±0.92) ％ in 48 h, respectively. The results indicated that the expression of VEGFR2 in SHI-1 cells treated with artesunate at different concentrations reduced significantly (P ＜0.05), but there was no significant difference between 24 h group and 48 h group (P ＞0.05). Conclusion The concentration of VEGF in SHI-1 cell was high, and artesunate can down-regulate the expression of VEGF and VEGFR2, but the effect of artesunate on the VEGFR1 was not significant.