芒柄花素对胰腺癌荷瘤小鼠局部免疫调节的作用及机制  

Effect and mechanism of formononetin mediated TLR4/iNOS pathway on local immune regu⁃lation in tumor bearing mouse model of pancreatic cancer

作　　者：常城 胡起茂 严红梅[1] 许丹[1] CHANG Cheng;HU Qi-mao;YAN Hong-mei;XU Dan(Wuhan Hospital of Traditional Chinese medicine,Wuhan 430071,China;Huangshi Hospital of Traditional Chinese medicine,Huangshi 435000,China)

机构地区：[1]武汉市中医医院,湖北武汉430071 [2]黄石市中医医院,湖北黄石435000

出　　处：《时珍国医国药》2024年第15期3377-3382,共6页Lishizhen Medicine and Materia Medica Research

基　　金：国家中医药管理局资助项目(国中医药人教函[2022]75号;国中医药人教函[2018]134号)。

摘　　要：目的探讨芒柄花素介导Toll样受体4(TLR4)/诱导型一氧化氮合酶(iNOS)通路对胰腺癌荷瘤小鼠局部免疫调节的作用。方法取50只BALB/c小鼠,采用腋下注射小鼠胰腺癌细胞MPC-83方式构建胰腺癌荷瘤小鼠模型。将建模成功小鼠随机分为模型组、吉西他滨组(0.5 mg/g)、芒柄花素低、中、高剂量组(25,50,100 g/kg),每组10只。给药结束3d后,计算抑瘤率;苏木素伊红染色观察肿瘤组织病理情况;Griess法检测一氧化氮(NO)含量;实时-定量聚合酶链反应检测TLR4、核转录因子-κB p65(NF-κB p65)、iNOS信使核糖核酸(mRNA)表达;蛋白质免疫印迹法检测TLR4、NF-κB p65、iNOS蛋白表达及p-NF-κB p65水平。结果与模型组比,吉西他滨组和芒柄花素3剂量组在第7、14、21、28天肿瘤体积均减少,肿瘤质量、NO含量、TLR4、NF-κB p65、iNOS mRNA和蛋白表达及p-NF-κB p65水平显著下降,差异有统计学意义(P<0.05或P<0.01)。结论芒柄花素可改善胰腺癌荷瘤小鼠局部免疫功能,可能与抑制肿瘤微环境TLR4/iNOS通路,下调NO含量有关。Objective To investigate the effect of formononetin mediated Toll-like receptor 4(TLR4)/inducible nitric oxide synthase(iNOS)pathway on local immune regulation in tumor bearing mouse model of pancreatic cancer.Methods 50 BALB/c mice were injected with mouse pancreatic cancer cells MPC-83 under the armpit to establish a tumor bearing mouse model of pancreatic cancer.The successfully modeled mice were randomly divided into model group,gemcitabine group(0.5 mg/g),low,medium and high dose formononetin groups(25,50 and 100 g/kg),with 10 in each group.3 days after the end of administration,the tumor inhibition rate was calculated,and hematoxylin eosin(HE)staining was used to observe the micropathology of tumor tissue,and the content of NO was determined by Griess method,and the expressions of TLR4,nuclear transcription factor-κB p65(NF-κB p65)and iNOS mRNA were detected by real-time quantitative polymerase chain reaction,and the expressions of TLR4,NF-κB p65,iNOS protein and the level of p-NF-κB p65 were detected by western blotting.Results Compared with model group,tumor volume in gemcitabine group and formononetin 3 dose groups were decreased on day 7,14,21 and 28,and tumor mass,the content of NO,and the mRNA and protein expressions of TLR4,NF-κB p65,iNOS and the level of p-NF-κB p65 in tumor tissue were significantly decreased,with statistically significant(P<0.05 or P<0.01).Conclusion Formononetin can improve the local immune function of pancreatic cancer bearing mice,which may be related to the inhibition of TLR4/iNOS pathway in tumor microenvironment,the down-regulation of NO content.

关 键 词：芒柄花素 TOLL样受体4 诱导型一氧化氮合酶 胰腺癌 局部免疫调节 

分 类 号：R285.5[医药卫生—中药学]