摘要
黄芪-莪术配伍是临床常用治疗肿瘤的组合,基于Th17/Treg细胞平衡探究黄芪-莪术配伍联合5-氟尿嘧啶(5-fluo-rouracil,5-FU)对结肠癌原位移植瘤模型小鼠肿瘤生长的可能作用机制。该研究将90只BALB/c雄性小鼠随机分为9组:空白组,模型组,5-氟尿嘧啶(5-FU)组,黄芪-莪术(2∶1)配伍高、中、低剂量组,联合5-FU高、中、低剂量组,每组10只。除空白组外其余各组小鼠建立CT26.WT结肠癌原位移植瘤模型。术后24 h空白组与模型组给予生理盐水灌胃(10 mL·kg^(-1),每日1次);5-FU组用5-FU腹腔注射(25 mg·kg^(-1),隔日1次);黄芪-莪术高、中、低剂量组分别用黄芪-莪术药液灌胃(12、6、3 g·kg^(-1),每日1次);联合高、中、低剂量组分别用黄芪-莪术相应剂量灌胃和5-FU腹腔注射,用药剂量和方法同上。各组均干预3周后处死小鼠取肿瘤组织,称瘤体质量并计算平均瘤重;检测各组小鼠脾组织CD4^(+)T淋巴细胞中辅助性T细胞17(Th17)/调节性T细胞(Treg)百分比变化趋势;检测转化生长因子-β(transforming growth factor-β,TGF-β)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、γ-干扰素(IFN-γ)、Smad4、N-钙粘蛋白(N-cadherin)、基质金属蛋白酶-7(matrix metalloproteinase-7,MMP-7)在血液中mRNA表达水平及肿瘤组织中蛋白表达水平。结果表明,与模型组比较,各给药组小鼠瘤体质量均降低(P<0.01);各给药组小鼠脾组织中CD4^(+)IL-17;均有不同程度降低(P<0.001),CD4^(+)Foxp^(3+)细胞比例均升高(P<0.001或P<0.05),表明Th17/Treg始终保持动态平衡,且联合用药组的效果优于其他组别;各给药组血液中TGF-β、TNF-α、IFN-γ、Smad4、N-cadherin、MMP-7 mRNA及原位移植瘤中TGF-β、TNF-α、IFN-γ、Smad4、N-cadherin、MMP-7蛋白的表达均不同程度降低(P<0.01或P<0.001),并且呈一定剂量依赖性。黄芪-莪术配伍联合5-FU可抑制CT26.WT结肠癌小鼠原位移植瘤的生长,其机制可能与维持体内Th17/Treg的动态平衡,下调TGF-β、TNF-α、IFN-γ、Smad4、N-cadherin、MMP-7肿瘤相关因子的表达水平有关。
Astragali Radix-Curcumae Rhizoma(AR-CR)is a combination commonly used in the clinical treatment of tumors.Based on the T helper 17(Th17)/regulatory T cell(Treg)balance,the present study explored the possible mechanism of AR-CR combined with 5-fluorouracil(5-FU)on the tumor growth of orthotopic xenograft model mice of colorectal carcinoma.Ninety male BALB/c mice were randomly divided into nine groups,i.e.,a blank group,a model group,a 5-FU group,high-,medium-,and low-dose AR-CR(2∶1)groups,and high-,medium-,and low-dose AR-CR+5-FU groups,with 10 mice in each group.The orthotopic xenograft model of CT26.WT colorectal carcinoma was induced in mice except those in the blank group.Twenty-four hours after the ope-ration,mice in the blank group and the model group received normal saline by gavage(10 mL·kg^(-1),once per day),and those in the 5-FU group received 5-FU by intraperitoneal injection(25 mg·kg^(-1),once every other day).Mice in the AR-CR groups received AR and CR decoctions by gavage(12,6,and 3 g·kg^(-1),once a day)and those in the combination groups received AR and CR decoctions and 5-FU(doses and administration methods were the same as above).After intervention for three weeks,all mice were sacrificed and tumor tissues were collected.The tumor mass was weighed and the average tumor weight was calculated.The changing trend of Th17/Treg(%)in the CD4^(+)T lymphocytes of the spleen tissues of the mice in each group was detected.The mRNA expression in the blood and protein expression in the tumor tissues of transforming growth factor-β(TGF-β),tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ),Smad4,N-cadherin,matrix metalloproteinase-7(MMP-7)were detected.The experimental results revealed that compared with the model group,the groups with drug intervention showed reduced tumor mass(P<0.01),decreased CD4^(+)IL-17;in the spleen tissues to varying degrees(P<0.001),and increased proportion of CD4^(+)Foxp^(3+)(P<0.001 or P<0.05),indicating that Th17/Treg maintained dynamic balance,and the effect of the combination groups was predominant.Additionally,the mRNA expression in the blood and protein expression in the tumor tissues of TGF-β,TNF-α,IFN-γ,Smad4,N-cadherin,and MMP-7 declined to varying degrees in a dose-dependent manner(P<0.01 or P<0.001).The AR-CR combined with 5-FU can inhibit the tumor growth of orthotopic xenograft model mice of CT26.WT colorectal carcinoma.The mechanism may be related to maintenance of Th17/Treg dynamic balance in the body and down-regulation of TGF-β,TNF-α,IFN-γ,Smad4,N-cadherin,and MMP-7 expression.
作者
郭文晖
齐卓操
关汉卿
刘甜甜
梁立
余倩慧
梁研
唐德才
GUO Wen-hui;QI Zhuo-cao;GUAN Han-qing;LIU Tian-tian;LIANG Li;YU Qian-hui;LIANG Yan;TANG De-cai(Shool of Traditional Chinese Medicine-School of Integrated Chinese and Western Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,China;Basic Medical College,Shanghai University of Chinese Medicine,Shanghai 201203,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2022年第1期167-175,共9页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(81873021)
江苏省中医药科技发展专项(2020ZX01)
江苏省研究生科研创新计划项目(KYCX20_1576)。
