丹参川芎嗪联合生脉注射液对肝星形细胞LX-2增殖和凋亡的影响

Effect of salvia miltiorrhizae and ligustrazine combined with Shengmai injection on proliferation and apoptosis of LX-2 cells

目的观察丹参川芎嗪注射液联合生脉注射液对肝星形细胞LX-2增殖和凋亡的影响。方法取对数生长期的LX-2细胞,接种于96孔板(1. 0×10~5/ml),分为实验组和空白对照组。实验组分别设置5个药物浓度梯度:丹参川芎嗪注射液(浓度为1,2,4,8,16μl/ml)和生脉注射液(浓度为0. 6,1. 2,2. 4,4. 8,6μl/ml)。药物作用24 h和48 h,用CCK-8法检测细胞增殖情况。选择丹参川芎嗪注射液1μl/ml和生脉注射液4. 8μl/ml联合作用于LX-2细胞24 h和48 h,用CCK-8法检测细胞增殖情况,用流式细胞仪检测用药24 h的细胞凋亡情况。结果丹参川芎嗪注射液作用24 h,除1,2μl/ml外,其余浓度对LX-2细胞的抑制率高于对照组(P <0. 05),抑制率分别为13. 21%,25. 74%,31. 86%。丹参川芎嗪作用48 h,除1μl/ml,其余浓度对LX-2细胞的抑制率高于对照组(P <0. 05),抑制率分别为16. 40%,28. 50%,31. 82%,37. 20%。生脉注射液作用LX-2细胞24 h,除0. 6,1. 2μl/ml外,其余浓度对LX-2细胞的抑制率高于对照组(P <0. 05),抑制率分别为14. 87%,20. 30%,31. 16%;药物作用48 h,除0. 6,1. 2μl/ml外,其余浓度对LX-2细胞的抑制率高于对照组(P <0. 05),抑制率分别为14. 08%,34. 13%,77. 82%。联合用药24 h,联合用药组和生脉注射液(SMI)组对LX-2细胞的抑制率高于空白对照组(P <0. 05),且联合用药的效果要好于丹参川芎嗪和生脉注射液单独用药(P <0. 05);联合用药48 h,联合用药组,生脉注射液(SMI)组和丹参川芎嗪(SML)组对LX-2细胞的抑制率高于空白对照组(P <0. 05),且联合用药的抑制率要高于丹参川芎嗪单独用药(P <0. 05)。结论一定浓度的丹参川芎嗪注射液和生脉注射液均能抑制LX-2细胞的增殖,联合用药24 h,其对LX-2细胞的抑制率高于丹参川芎嗪注射液和生脉注射液单独用药。

Objective To investigate the effect of salvia miltiorrhizae and ligustrazine(SML) combined with Shengmai injection(SMI) on the proliferation and apoptosis of LX-2 cells. Methods LX-2 cells in logarithmic growth phase were seeded in 96- well plates(1.0×10 5/ml). LX-2 cells were divided into experimental groups and blank control group. LX-2 cells were treated with SML(1, 2, 4, 8, 16 μl/ml) and SMI(0.6, 1.2, 2.4, 4.8, 6 μl/ml) to screen the best concentration,respectively. The proliferation of LX-2 cells was detected by CCK-8 method at 24 h and 48 h. Then, SML(1 μl/ml) combined with SMI(4.8 μl/ml) was applied to treat LX-2 cells, and then cell proliferation was measured by CCK-8 at 24 h and 48 h. Flow cytometry was used to detect cell apoptosis at 24 h. Results Cell proliferation was significantly suppressed by SML(4,8,16 μl/ml) at 24 h( P <0.05),and the inhibition rates were 13.21%, 25.74% and 31.86%,respectively. After 48 h treatment, except 1 μl/ml, the other concentrations of SML inhibited the proliferation of LX-2 cells( P <0.05), and the inhibition rates were 16.40%, 28.50%, 31.82% and 37.20%,respectively. Except 0.6 and 1.2 μl/ml SML, the inhibition rate of the other concentrations of SMI was higher than that of blank control at 24 h( P <0.05), and the inhibition rates were 14.87%, 20.30% and 31.16%,respectively. After 48 h treatment, except 0.6 and 1.2 μl/ml, the other concentrations of SMI inhibited the proliferation of LX-2 cells( P <0.05), and the inhibition rates were 14.08%, 34.13% and 77.82%,respectively. Combined therapy for 24 h, the inhibitory rates were significantly higher in combination group and Shengmai injection(SMI) group than in control group( P <0.05), and the effect of combination therapy was better than that of Danshen ligustrazine and Shengmai injection alone( P <0.05). After treatment for 48 h, the inhibitory rates were significantly higher in combination group, SML group and SMI group than in control group( P <0.05), and the inhibitory effect of drug combination was better than that of salvia miltiorrhizae and ligustrazine alone( P <0.05). Conclusion An effective concentration of Salvia miltiorrhizae and ligustrazine injection and Shengmai injection can inhibit the proliferation of LX-2 cells. The inhibitory effect of their combination is better than that of single drug at 24 h.