益气解毒方调节线粒体途径阻断气虚染毒大鼠鼻咽上皮细胞癌变发生

Intervening Effect of Qi-Boosting Toxin-Resolving Formulae in the Developing Process of Nasopharyngeal Carcinogenesis among Rats by Regulating Mitochondrion Pathway

目的探讨益气解毒方通过调节线粒体途径阻断气虚染毒大鼠鼻咽癌变的发生。方法 72只SD大鼠随机分为模型组、益气解毒方组和维甲酸组。腹腔注射二亚硝基哌嗪(DNP)和佛波醇酯(TPA),同时进行力竭游泳,建立大鼠鼻咽癌变模型,益气解毒方组和维甲酸组分别给予相应药物进行干预治疗,定期分批处死大鼠,取鼻咽组织做病理检测,免疫组织化学SABC法检测蛋白表达,RT-PCR检测mRNA的表达。结果在诱发性鼻咽癌变进程的单纯性增生阶段,与模型组比较,益气解毒方组和维甲酸组大鼠鼻咽上皮细胞的Bcl-2、Bax、Cytochrome-C和Caspase-3蛋白表达差异均无统计学意义(P>0.05)。而在异型增生阶段,益气解毒方组鼻咽上皮细胞的Bcl-2蛋白表达活性显著低于模型组(P<0.05),Cytochrome-C和Caspase-3又分别显著高于模型组(P<0.05);Bax蛋白表达活性虽然高于模型组,但比较分析结果表明,组间差异却无统计学意义(P>0.05)。在同一病理阶段,各组鼻咽上皮细胞的Bax和Bcl-2 mRNA转录活性与各自蛋白的表达活性呈现平行趋势。结论益气解毒方能够有效阻逆大鼠鼻咽上皮细胞癌变进程,其作用与其有效调节线粒体途径关键基因的表达密切相关。

Objective To investigate the effects of Qi-Boosting Toxin-Resolving Formulae （QBTRF） by regulating mitochondrion pathway in the developing process of nasopharyngeal carcinogenesis among rats. Methods 72 SD rats were randomly divided into model group, Qi-Boosting Toxin-Resolving Formulae （QBTRF） group, vitamin A acid group. All these animals were subcutaneously injected in axillary fossa with N, N＇dinitrosopiperazine （DNP） and 12-O-tetradecanoylphorbol （TPA） in combination with exhausted swimming to establish the model of nasopharyngeal carcinomas. QBTRF was given to intervention. Samples were fixed in 4% neutral paraformaldehyde for pathohistological evaluation, and the protein expression were detected by SABC immunohistochemical and mRNA expression were detected by RT-PCR. Result In the simple hyperplasia stage of induced cancerous transforming lesion in the nasopharyngeal epithelia, no significant difference were determined in the expressing activities of Bcl-2, Bax, Cytochrome-C and Caspase-3 in QBTRF group and vitamin A acid group （P〉0.05）, compared with the model group respectively. At the pathological stage of atypical hyperplasia, the expressing activity of Bel-2 was significantly lower （P 〈0.05） and the activities of Cytochrome-C and Caspase-3 were significantly higher in QBTRF group than that of model group （P〈0.05）. Though the expressing activity of Bax was higher in QBTRF group than that of model group, no significant difference was determined between them （P 〉 0.05）. There was a parallel tendency of expressing activity in either Bax mRNA or Bel-2 mRNA with Bax or Bcl-2 itself respectively in any stage of pathology. Conclusion The anticancer effect of QBTRF was realized by regulating mitochondrion pathway in the developing process of carcinogenesis among rats.