基于Nrf2信号通路探讨艾迪注射液联合GP化疗干预Lewis肺癌小鼠的作用研究

The effect of Aidi injection combined with GP chemotherapy on Lewis lung cancer mice based on Nrf2 signaling pathway

目的探讨艾迪注射液联合吉西他滨+顺铂化疗干预Nrf2信号通路对Lewis肺癌小鼠的作用。方法建立Lewis肺癌小鼠模型40只,随机分为5组,模型组每天给予生理盐水0.1 mL/10 g;GP化疗组给予顺铂5 mg/kg+吉西他滨50 mg/kg;低、中、高剂量组在GP化疗组的基础上分别给与艾迪注射液0.05 mL/10 g、0.1 mL/10 g、0.2 mL/10 g。顺铂每5天给予一次,吉西他滨每7天给予一次,给药共14 d。实验期间观察小鼠的一般活动状况,监测小鼠体质量。实验结束后检测小鼠瘤体积;ELISA法检测IL-6表达水平;免疫组化法检测Nrf2信号通路关键蛋白的表达;HE染色分析小鼠各主要脏器及肿瘤组织的病理变化。结果(1)治疗14 d后,与模型组比较,0~5 d各治疗组小鼠体质量均出现不同程度下降,而随着实验时间的延长,低、中和高剂量组小鼠体质量开始增长,而GP化疗组小鼠后期体质量呈下降趋势。与模型组比较,各治疗组小鼠瘤体积显著降低(P<0.01),并且各组瘤重显著下降(P<0.001),其中中剂量组的抑瘤率最高,高剂量组及低剂量组次之,GP化疗组抑瘤率最低。(2)与模型组比较,GP化疗组脾与胸腺指数显著降低(P<0.05),而与GP化疗组比较,低、中和高剂量组脾与胸腺指数显著增加。(3)与模型组比较,各组IL-6表达均明显下降(P<0.05);与GP化疗组相比,中剂量组IL-6表达明显下降(P<0.05)。(4)与模型组比较,GP化疗导致的肝、肾脏器损伤可被艾迪注射液改善,并能协同GP化疗杀伤肿瘤细胞。(5)与模型组相比,GP化疗组Nrf2、NQO1、GSTA1和HO-1蛋白表达量均上升(P<0.05);与GP化疗组相比,低、中和高剂量组Nrf2、NQO1、GSTA1和HO-1蛋白表达量均降低(P<0.05)。结论艾迪注射液联合GP化疗可干预Nrf2信号通路发挥抗小鼠Lewis肺癌作用,且艾迪注射液可对GP化疗起到减毒增效作用。

Objective To investigate the effect of Aidi injection combined with gemcitabine and cisplatin chemotherapy on Nrf2 signaling pathway in Lewis lung cancer mice.Methods Forty Lewis lung cancer model in mice were established,which were randomly divided into five groups:model group was given normal saline(0.1 mL/10 g)every day.GP chemotherapy group was given cisplatin 5 mg/kg+gemcitabine 50 mg/kg.On the basis of GP chemotherapy group,the low-dose,medium-dose and high-dose groups were given 0.05 mL/10 g,0.1 mL/10 g,0.2 mL/10 g Aidi injection respectively.Cisplatin was given every 5 days,and gemcitabine was given every 7 days for 14 days.The general activity of the mice was observed and the weight and the change of the tumors volume in mice were recorded.The expression level of IL-6 was detected by ELISA,and expression of key proteins in Nrf2 signaling pathway were detected by immunohistochemical staining,the pathological changes of the major organs and the tumors in mice were tested by HE staining.Results(1)After 14 days of treatment,the body weight of mice in each treatment group was decreased in different degrees compared with model group in 0-5 days.With the extension of experimental time,the body weight of mice in low-dose,medium-dose and high-dose groups began to increase,while the body weight of mice in GP chemotherapy group showed a downward trend in the later stage.Compared with the model group,tumor volume of each treatment group was significantly reduced(P<0.01),and tumor weight of each treatment group was significantly reduced(P<0.001),the medium-dose group had the highest tumor inhibition rate,followed by the high-dose and low-dose groups,and the GP chemotherapy group showed the lowest tumor inhibition rate.(2)Compared with the model group,the index of spleen and thymus in GP chemotherapy group decreased significantly(P<0.05),compared with GP chemotherapy group,the index of spleen and thymus in low-dose,medium-dose and high-dose groups increased significantly(P<0.05).(3)Compared with model group,the expression of IL-6 in each group decreased significantly(P<0.05).Compared with GP chemotherapy group,the expression of IL-6 in the medium-dose group decreased significantly(P<0.05).(4)Compared with model group,the damage of liver and kidney organs caused by GP chemotherapy could be improved by Aidi injection,and tumor cells could be killed with GP chemotherapy.(5)Compared with model group,the expression levels of Nrf2,NQO1,GSTA1 and HO-1 protein in GP chemotherapy group were higher than those in model group(P<0.05).Compared with GP chemotherapy group,the expression levels of Nrf2,NQO1,GSTA1 and HO-1 protein in low-dose,medium-dose and high-dose groups decreased(P<0.05).Conclusion Aidi injection combined GP chemotherapy could play the role on reducing toxicity and increasing efficacy through intervening Nrf2 signaling in anti-Lewis lung cancer.