小柴胡汤对C6胶质瘤大鼠ESM-1、VEGF表达的实验研究

The experimental research of ESM-1 and VEGF expression in Xiaochaihutang on C6 rat glioma

目的探讨小柴胡汤对C6胶质瘤大鼠ESM-1和VEGF蛋白表达水平的影响。方法清洁级Wistar老年雄性大鼠18月龄,n=60只,右前肢腋窝皮下接种2×106/20μl的细胞悬液;然后随机分为6组:肿瘤对照组、阳性药A组(替莫唑胺;50mg/Kg)、阳性药B组(替莫唑胺;25mg/Kg)、小柴胡汤C组(50g/Kg)、小柴胡汤D组(20g/Kg)、小柴胡汤E组(10g/Kg);每组均分别灌胃给予生理盐水、阳性药A、阳性药B、小柴胡汤C、小柴胡汤D和小柴胡汤E,连续服药5d,休息4d,共给药3次,于最后一次给药后2d处死动物,眼球取血及取肿瘤组织。采用ELISA酶联免疫法检测大鼠血清及肿瘤组织中ESM-1和VEGF的蛋白含量;采用免疫组织化学方法检测大鼠肿瘤组织中ESM-1和VEGF的蛋白表达水平。结果与肿瘤对照组比较,小柴胡汤各剂量组大鼠血清及肿瘤组织中ESM-1、VEGF蛋白含量和蛋白表达水平均降低,蛋白含量随剂量增长降低趋势明显差异有统计学意义(P<0.01),蛋白表达水平C组、D组降低较为显著但二者之间差异无统计学意义(P>0.05);阳性药各剂量组大鼠血清及肿瘤组织中ESM-1、VEGF蛋白含量和蛋白水平亦均降低,且随剂量增长降低趋势明显差异亦有统计学意义(P<0.01);阳性药A组、小柴胡汤C组和D组这三组之间在肿瘤组织蛋白表达水平上差异无统计学意义(P>0.05)。结论与阳性药各剂量组的治疗机制相一致,小柴胡汤各剂量组均能通过下调ESM-1和VEGF的水平治疗C6胶质瘤,高剂量组作用最为显著,其作用机制有待于进一步探讨。

Objective To investigate Xiaochaihutang impact on ESM-1 and VEGF protein expression in C6 glioma in rats.Methods Wistar male rats aged 18 months,n=60,with the right forelimb axillary subcutaneous inoculation of cell suspension 2＆#215;106/20 μl;then randomly divided into six groups ：control group tumors ,positive drug group A（temo-zolomide;50 mg/Kg）,positive drug group B（temozolomide;50 mg/Kg）,Xiaochaihutang group C（50 mg/Kg）,Xi-aochaihutang group d（20 mg/Kg）,Xiaochaihutang group E（10 mg/Kg）;each group were orally administered saline, positive drug A,positive drug B,Xiaochaihutang C,Xiaochaihutang D and E,continuous medication 5 d,rest 4 d,were to drug three times in the last 2 d animals were sacrificed after administration ,blood and tumor tissue.Using ELISA to detect the protein content of ESM-1 and VEGF in serum and tumor tissue,Immunohistochemistry was used to detect the protein expression of ESM-1 and VEGF in rat tumor tissue.Results Compared with the control group tumors,the protein content and protein expression of ESM-1 and VEGF in Xiaochaihutang each dose group on serum and tumor tis-sue were decreased ,protein content reduced the growth trend with dose significant difference was statistically（P 〈0.01）;protein expression levels group C,D group decreased more significant but the difference between them was not statistically sifnificant（P〉0.05）;positive serum and tumor tissue of rats in each dose group in protein content and protein levels of ESM-1 and VEGF were lower,and with lower doses of growth trend evident difference was also statis-tically significant（P 〈0.01）;positive drug group A,Xiaochaihutang group C and D between the three groups ,the difference was not statistically significant（P〉0.05）.Conclusion Consistent with mechanism of positive drug treat-ment each dose group ,Xiaochaihutang each dose group could cut through the level of ESM-1 and VEGF treatment C6 glioma high-dose group the most significant effect,the mechanism needs further explored.