文章摘要
王硕,廖文宇,程权,等.香草扶正合剂对Lewis肺癌模型小鼠癌细胞自噬的影响[J].浙江中医药大学学报,2020,44(12):1152-1159.
香草扶正合剂对Lewis肺癌模型小鼠癌细胞自噬的影响
Effect of Xiangcao Fuzheng Mixture on Autophagy of Lewis Lung Cancer Mice
DOI:10.16466/j.issn1005-5509.2020.12.003
中文关键词: 细梗香草  香草扶正合剂  顺铂  肺癌  自噬  自噬相关蛋白  凋亡
英文关键词: Lysimachia capillipes Hemsl  Xiangcao Fuzheng Mixture  cisplatin  lung cancer  autophagy  autophagy-related protein  apoptosis
基金项目:杭州市卫生科技计划(重点)项目(2018Z02);浙江省中医药防治重大疾病攻关计划项目(2018ZY009)
作者单位E-mail
王硕 浙江中医药大学第四临床医学院 杭州 310053  
廖文宇 浙江中医药大学第四临床医学院 杭州 310053  
程权 浙江大学医学院附属杭州市第一人民医院  
丁志山 浙江中医药大学医学技术学院  
傅华洲 浙江大学医学院附属杭州市第一人民医院 fuhuazhou@126.com 
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中文摘要:
      [目的]探讨香草扶正合剂(Xiangcao Fuzheng Mixture,XCFZ)的抑瘤作用及其机制。[方法]建立Lewis肺癌小鼠移植瘤模型,随机分为模型组,XCFZ低、中、高剂量组,顺铂组(阳性对照组),并设置空白组。模型组以双蒸水0.2mL·d-1灌胃,XCFZ低、中、高剂量组分别以XCFZ低、中、高剂量0.2mL·d-1灌胃,XCFZ低、中、高剂量浓度分别为0.93g·mL-1、1.86g·mL-1、3.72g·mL-1;顺铂组以0.64mg·mL-1的顺铂溶液0.1mL·(2d)-1腹腔注射,同时以双蒸水0.2mL·d-1灌胃。每天固定时间给药,给药14d。称取小鼠体质量、瘤质量和去瘤后体质量,并计算抑瘤率,常规石蜡切片观察肿瘤组织病理改变,透射电镜观察瘤组织自噬小体,Western blot检测瘤组织自噬相关基因Beclin1、自噬相关蛋白5(autophagy related 5,Atg5)、微管相关蛋白1轻链3-β(microtubule associated protein 1 light chain 3-β,LC3b)蛋白表达水平,实时荧光定量PCR技术(Real-time quantitative PCR,RT-qPCR)检测瘤组织Beclin1、Atg5、LC3b、Unc-51样自噬激活激酶1(Unc-51 like autophagy activating kinase 1,ULK1)、B细胞淋巴瘤-2(B-cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)mRNA表达水平。[结果]与模型组比较,各给药组瘤质量降低(P<0.01)。顺铂组抑瘤率最高,其次为XCFZ中剂量组。与模型组比较,XCFZ中剂量可升高小鼠去瘤后体质量(P<0.05),增加肿瘤组织Atg5、Beclin1、LC3b蛋白及Atg5、Beclin1、LC3b、ULK1 mRNA表达水平,并升高Bax/Bcl-2 mRNA比值(P<0.05,P<0.01)。与顺铂组比较,XCFZ中剂量可升高小鼠去瘤后体质量(P<0.01),增加肿瘤组织Atg5、LC3b蛋白及Beclin1 mRNA表达水平(P<0.05,P<0.01,P<0.05)。[结论]XCFZ对肺癌小鼠模型肿瘤生长具有抑制作用,其抑瘤作用可能与自噬诱导相关;或与增加自噬相关蛋白Beclin1、Atg5表达,引发凋亡相关。
英文摘要:
      [Objective] To explore the anti-tumor effect and mechanism of Xiangcao Fuzheng Mixture(XCFZ). [Methods] Lewis lung cancer mouse transplanted tumor model was successfully established and the mice were randomly divided into model group, XCFZ low, medium and high dose groups, cisplatin group(positive control group) and blank group. Mice in model group were given double distilled water 0.2mL·d-1; XCFZ low, medium and high dose groups were given 0.2mL·d-1 of XCFZ low, medium and high dose, and the low, medium and high dose concentrations of XCFZ were 0.93g·mL-1, 1.86g·mL-1 and 3.72g·mL-1, respectively; while cisplatin group was given 0.64mg·mL-1 cisplatin solution 0.1mL·(2d-1) of intraperitoneal injection, and double distilled water 0.2mL·d-1. It was administered at a fixed time every day for 14 days. The body mass, tumor mass and body mass were weighed after tumor removal of mice, and tumor inhibition rate was calculated. Conventional paraffin sections were used to observe pathological changes of tumor tissues. The autophagosome of tumor was observed by transmission electron microscope. The protein expression levels of Beclin1, autophagy related 5(Atg5) and microtubule associated protein 1 light chain 3-β(LC3b) in tumor tissues were detected with Western blot. Real-time quantitative PCR was used to detect the mRNA expression levels of Beclin1, Atg5, LC3b, Unc-51 like autophagy activating kinase 1(ULK1), B-cell lymphoma-2(Bcl-2) and Bcl-2 associated X protein(Bax) in tumor tissues. [Results] Compared with model group, the tumors mass was decreased in each administration group(P<0.01). Cisplatin group had the highest tumor inhibition rate, followed by XCFZ medium dose group. Compared with model group, XCFZ medium dose group could increase the weight of mice after tumor removal(P<0.05), and increase the protein expression levels of Atg5, Beclin1, LC3b, and mRNA expression levels of Atg5, Beclin1, LC3b, ULK1 in tumor tissues, and increase the ratio of Bax/Bcl-2 mRNA(P<0.05,P<0.01). Compared with cisplatin group, XCFZ medium dose group can increase the weight of mice after tumor removal(P<0.01), and increase the expression levels of Atg5, LC3b protein and Beclin1 mRNA in tumor tissues(P<0.05, P<0.01, P<0.05). [Conclusion] XCFZ can inhibit tumor growth in lung cancer mouse, which may be related to autophagy induction, or to increasing the expression of autophagy-related proteins Beclin1 and Atg5 and inducing apoptosis.
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