丹参注射液对自杀基因tk/GCV系统的协同增效作用

目的探讨丹参注射液联合自杀基因系统对大鼠肝癌细胞和小鼠移植瘤的作用。方法①丹参注射液以不同浓度与GCV分别或共同作用于大鼠肝癌CBRH7919的tk-细胞,以及含5%tk+的tk+、tk-混合细胞,MTT法检测各组存活率,用Q值(实测药效与理论药效的比值)分析中药与自杀基因系统联合的相互作用是否有协同性,0.85≤Q<1.15为相加;Q≥1.15为协同。②把小鼠肝癌细胞H22/tk(tk+)与H22(tk-)细胞按1∶4混合,再按2×106细胞/只接种到Balb/c纯系小鼠腋下皮下组织内,随机分为模型组、丹参(注射液)组、tk/GCV组、tk/GCV联合丹参注射液治疗的联合组,n=9~11;中药治疗从d 2起共15 d,自杀基因系统治疗从长出肿瘤后开始共10 d,进行疗效观察。结果①各组存活率tk/GCV组为63.10%±2.17%,GCV组为67.03%±2.81%,5 ml.L-1丹参联合tk/GCV组为26.67%±4.23%(Q=1.22),5 ml.L-1丹参+GCV组为42.17%±5.36%(Q=1.04);10 ml.L-1丹参联合tk/GCV组为18.10%±5.56%(Q=1.26),10 ml.L-1丹参+GCV组为33.84%±9.84%(Q=1.06)。②接种后各组均在d 7触摸到肿瘤,成瘤率100%。联合组在治疗后肿瘤生长呈现明显抑制,最终肿瘤体积(1.53±0.88)cm3,比模型组(3.19±1.76)cm3减少52.01%(P<0.05);肿瘤质量(1.32±0.80)g,比模型组(2.28±1.20)g减少42.11%(P<0.05),差异有统计学意义。丹参组、tk/GCV组肿瘤体积、瘤块质量与模型组比较差异均无统计学意义。结论丹参注射液能提高自杀基因系统对大鼠肝癌细胞的杀伤力和对小鼠肝癌细胞移植瘤的抑制作用,两者联合作用的协同性提示丹参注射液能增强自杀基因旁观者效应。

The synergistic effect of Danshen injection combining with suicide gene tk/GCV system

Aim To investigate the curative effect of Danshen injection combining with HSV-tk/GCV system on rats′ hepatocarcinoma cells and murine transplanted hepatocarcinoma.Methods ① Rats′ hepatocarcinoma cell line CBRH7919(tk~-),CBRH7919/tk(tk~+) and the 5% tk~+ mixed cells were treated with diverse concentrations of Danshen injection,GCV separately,and Danshen injection plus GCV(n=3).The survival rate of each groups was examined using MTT Assay and was analyzed using paired comparison.Q-value analysis method was used to estimate the synergistic effect of Chinese herbal on the suicide gene system.Q-value is a ratio of the actural effect of combination treatment to its theoretical effect.It is thought to be an additive effect when 0.85≤Q<1.15 and a synergistic effect when Q≥1.15.②Four experimental groups were model control group,Danshen injection group;tk/GCV group and the combining group(tk/GCV+Danshen injection).The cells tk~+ and tk~-were mixed in the proportion of 1 to 4 and then were transplanted subcutaneously into murine oxter.The injection amount was 2×10~6 cells per mouse.The Danshen injection group and the combining group had been treated with Danshen injection from d 2 on for 15 d,while the tk/GCV group and the combining group had been treated with GCV when the tumor had been detectable.The curative effect was observed.Results ① When the concentration of GCV was 0.039 mmol·L~(-1),the survival rate was(63.10±2.17)% in tk/GCV group and(67.03±2.81)% in GCV group.While the survival rate was (26.67±4.23)%(Q=1.22)in the group of 5 ml·L~(-1) Danshen injection combining with tk/GCV,(42.17±5.36)%(Q=1.04)in the group of 5 ml·L~(-1) Danshen injection combining with GCV,and(18.10±5.56)%(Q=1.26)in 10 ml·L~(-1) Danshen injection combining with 5% tk~+/GCV,(33.84±9.84)%(Q=1.06)in the group of 10 ml·L~(-1) Danshen injection combining with GCV. ②The tumor bumps could be touched in mice of every group 7 d after inculation.With the treating of tk/GCV system combined with Danshen injection,the tumor growth speeds stepped down,and the average dimension of tumor was(1.53±0.88) cm~3,decreased 52.01% in comparison with the control group′s(3.19±1.76) cm~3,P<0.05),and the average weight of tumor was(1.32±0.80) g,42.11% less than the control groups(2.28±1.20) g(P<0.05),significantly different in statistics.Although the average tumor size and the tumor weight in the Danshen injection remedy group and in the tk/GCV group were respectively less than that in the control group,there was no significant difference in statistics.Conclusion Danshen injection is capable of increasing the effects cooperating with the tk suicide gene system.Its synergistic effect alluded Danshen injection can improve the bystander effect of suicide gene therapy.