抗癌解毒开窍方抗脑胶质瘤的作用及机制研究

Study on the Effect and Mechanism of Kang-ai Jiedu Kaiqiao Decoction against Glioma

目的观察抗癌解毒开窍方抗脑胶质瘤的作用及其调控凋亡及耐药机制。方法通过构建裸鼠皮下胶质瘤模型,将24只SPF级BALB/c成瘤裸鼠按随机数字表法分为模型组(生理盐水0.3 ml,24 d)、替莫唑胺组[TMZ 50 mg/(kg·d),6 d]、抗癌解毒开窍方组[抗癌解毒开窍方517 mg/(20 g·d),24 d]和联合组[TMZ 50 mg/(kg·d),6 d+抗癌解毒开窍方517 mg/(20 g·d),24 d],每组各6只,每天灌胃给药。给药期间,每3 d记录1次小鼠体质量与肿瘤体积。采用Westernblot法进行凋亡相关蛋白(Bax、Bcl-2)和耐药性相关蛋白(MGMT)水平检测,HE染色法观察瘤组织病理形态。结果肿瘤体积测量结果提示,与对照组比较,替莫唑胺和抗癌解毒开窍方及联合用药对脑胶质瘤的生长增殖均具有抑制作用,差异有统计学意义(P<0.05)。WB检测结果显示,与对照组比较,抗癌解毒开窍方组和TMZ组的Bax/Bcl-2比值升高,差异有统计学意义(P<0.05),两药联合组的Bax/Bcl-2比值明显升高,差异有统计学意义(P<0.05);与TMZ组比较,抗癌解毒开窍方组和两药联合组的Bax/Bcl-2比值升高,差异有统计学意义(P<0.05)。与对照组比较,TMZ组MGMT蛋白表达增加,差异有统计学意义(P<0.05);与TMZ组比较,抗癌解毒开窍方组和两药联合组MGMT蛋白表达降低,差异有统计学意义(P<0.05)。HE染色结果显示联合用药组相较模型组和单独用药组,细胞排列较整齐,细胞形态较为均一。结论抗癌解毒开窍方具有抑制胶质瘤生长增殖作用,其作用与上调Bax/Bcl-2比值有关。同时可抑制MGMT表达,在一定程度上减缓替莫唑胺耐药。

Objective To investigate the effect of Kang-ai Jiedu Kaiqiao Decoction(KAJDKQF)on the apopto-sis of glioma and drug resistance mechanism.Method Twenty-four SPF-grade BALB/c mice were used to construct subcutaneous glioma model.The tumor bearing mice were randomly divided into the following groups:model group(saline 0.3 ml,24 d),temozolomide group(TMZ 50 mg/(kg·d),6 d),KAJDKQF group(KAJDKQF 517 mg/(20 g·d),24 d)and combination group(TMZ 50 mg/(kg·d),6 d+KAJDKQF 517 mg/(20 g·d),24 d),with 6 nude mice in each group.The drugs were administered by gastric gavage daily.Mouse body weight and tumor volume were measured every three days during the experiment.Western blot was conducted to detect the levels of apoptosis-associated proteins(Bax,Bcl-2)and drug resistance-associated protein(MGMT).Histopathological morphology of the tumor tissue were detected by HE staining.Results The results of tumor volume measurements indicated that compared with the control group,TMZ,KAJDKQF,and the combination therapy all exerted inhibitory effects on the growth and proliferation of gliomas,with statis-tically significant differences(P<0.05).Western blot results showed that compared with the control group,the Bax/Bcl-2 ratio was increased in the KAJDKQF group and the TMZ group(P<0.05).The combination group also showed a signifi-cantly increased Bax/Bcl-2 ratio compared to the control group(P<0.05).Compared with the TMZ group,the KAJD-KQF group and the combination group exhibited an elevated Bax/Bcl-2 ratio(P<0.05).The expression of MGMT pro-tein was significantly increased in the TMZ group as compared with the control group(P<0.05).Compared with the TMZ group,the KAJDKQF group and the combination group exhibited a significantly reduced expression of MGMT(P<0.05).The results of HE staining showed that compared with the model group and the monotherapy groups,the combination group had more orderly cell arrangement and more uniform cell morphology.Conclusion KAJDKQF has an inhibitory effect on the growth and proliferation of gliomas,which is related to the up-regulation of Bax/Bcl-2 ratio.Additionally,KAJDKQF inhibits the expression of MGMT and reduces temozolomide resistance to some extent.