丹参酮ⅡA对膀胱癌动物模型丝裂原活化蛋白激酶/细胞外调节蛋白激酶途径的影响

Effect of tanshinone ⅡA on p38mitogen-activated protein kinase/extracellular signal-regulated kinase pathway in bladder cancer animal model

目的观察丹参酮ⅡA(TanⅡA)通过调节丝裂原活化蛋白激酶(p38MAPK)/细胞外调节蛋白激酶(ERK)途径对膀胱癌动物模型肿瘤增殖及肿瘤组织内凋亡相关因子的影响。方法选择30只4~5周龄的雄性BALB/-nu裸鼠,购自北京维通利华科技服务有限公司,其中20只注射1×10^7/ml的T24细胞悬液,待肿瘤体积大于0.1 cm^3,根据随机数表法对其进行分组干预,共分为模型组、治疗组,每组10只,另外10只裸鼠作为对照组;模型组及对照组给予生理盐水腹腔内注射,治疗组给予15 mg/kg的TanⅡA腹腔注射,均干预2周;开始干预后每天测量瘤体积1次,绘制肿瘤增殖曲线并在干预结束后将裸鼠处死后取0.1 g肿瘤组织检测半胱氨酸蛋白酶3(Caspase-3)、脱氧核糖核酸修复酶降解产物(Cleaved PARP)、细胞凋亡调控因子(bax)、B淋巴细胞瘤-2基因蛋白(bcl-2)、丝裂原活化蛋白激酶(p38 MAPK)、磷酸化丝裂原活化蛋白激酶(p-p38 MAPK)、细胞外调节蛋白激酶(ERK)、磷酸化细胞外调节蛋白激酶(p-ERK)水平,采用卡方分析计数资料差异,LSD-t检验及方差检验分析组间或多组间数据差异。结果给药7 d后治疗组裸鼠瘤体积明显低于模型组,且差异有统计学意义(t=4.445,P<0.01);3组小鼠组织中Caspase-3、Cleaved PARP、bax、bcl-2水平差异有统计学意义(F=5.937、4.264、6.135、5.364,P<0.01);相较于模型组,治疗组小鼠组织中Caspase-3、bcl-2明显降低(0.54±0.10、0.56±0.12比0.87±0.12、0.88±0.19),Cleaved PARP、bax明显升高(0.94±0.21、0.79±0.13比0.65±0.12、0.32±0.08),且差异存统计学意义(t=6.681、4.503、3.392、9.731,P<0.01);3组小鼠组织中p-p38 MAPK、p-ERK水平差异,有统计学意义(F=3.046、3.216,P<0.05),相较于模型组,治疗组下小鼠组织中p-p38 MARK及p-ERK水平明显降低(0.61±0.11、0.54±0.11比0.87±0.13、0.69±0.13),且差异存统计学意义(t=4.828、4.503,P<0.01)。结论TanⅡA通过有效调节p38MAPK/ERK途径实现抑制膀胱癌动物模型肿瘤增殖并调控肿瘤组织内凋亡相关因子。

Objective To analyze the effect of tanshinoneⅡA(TanⅡA)on tumor proliferation and apoptosis-related factors in tumor tissues by regulating p38 mitogen-activated protein kinase(MAPK)/extracellular signal-regulated kinase(ERK)pathway.Methods Thirty male BALB/-Nu nude mice aged 4-5 weeks were selected and purchased from Beijing Weitong Lihua Technology Service Co.,Ltd.20 of them were injected with 10^7/ml T24 cell suspension until the tumor volume was greater than 0.1 According to the random number table method,the rats were divided into model group and treatment group with 10 rats in each group,and the other 10 nude mice were used as control group;the model group and control group were given intraperitoneal injection of normal saline,and the treatment group were given 15 mg/kg TanⅡa intraperitoneal injection for 2 weeks.After the intervention,the tumor volume was measured once a day,and the tumor proliferation curve was drawn.After the intervention,0.1 g tumor tissue was taken to detect Caspase-3,cleaved PARP,bax,bcl-2 and p38 The levels of MAPK,p-p38 MAPK,ERK and p-ERK were analyzed by chi square analysis,LSD-t test and variance test.Results After 7 days of administration,the tumor volume of the treatment group was significantly lower than that of the model group,and the difference was statistically significant(t=4.445,P<0.01)The levels of PARP,bax and bcl-2 in the treatment group were significantly lower than those in the model group(0.54±0.10,0.56±0.12 vs.0.87±0.12,0.88±0.19),respectively PARP and bax were significantly increased(0.94±0.21,0.79±0.13 vs.0.65±0.12,0.32±0.08),and the difference was statistically significant(t=6.681,P<0.01;t=4.503,P<0.01;t=3.392,P<0.01;t=9.731,P<0.01);p-p38 in the tissues of the three groups was significantly higher than that in the control group Compared with the model group,the levels of p-p38 mark and p-ERK in the treatment group were significantly lower than those in the model group(0.61±0.11,0.54±0.11 vs.0.87±0.13,0.69±0.13),and the difference was statistically significant(t=4.828,P<0.01;t=4.503,P<0.01).Conclusion TanⅡA can effectively inhibit the proliferation of bladder cancer animal models and regulate apoptosis-related factors in tumor tissues by effectively regulating the p38MAPK/ERK pathway.