基于PI3K/AKT/mTOR信号通路探讨和枢消积丸对H22肝癌荷瘤小鼠肿瘤生成的影响及作用机制  

Exploring the effects and mechanism of action of HXP on H22hepatocellular tumor-bearingmice tumorigenesis based on PI3K/AKT/mTOR signaling pathway

作　　者：谈雅芝 尹玥 彭孟云 汪静 TAN Yazhi;YIN Yue;PENG Mengyun(College of integrated traditional Chinese and Western Medicine of Southwest Medical University/Affiliated Hospital of Traditional Chinese Medicine of Southwest Medical University,Luzhou Sichuan 646000,China)

机构地区：[1]西南医科大学中西医结合学院,四川泸州646000 [2]西南医科大学附属中医医院,四川泸州646000

出　　处：《四川中医》2024年第8期52-57,共6页Journal of Sichuan of Traditional Chinese Medicine

基　　金：四川省科技厅项目(编号:2022YFS0619);四川省中医药管理局中医药科研专项(川中医药办发[2021]13号);西南医科大学附属中医医院国家中医临床研究基地建设单位科研项目(西南医大中医院[2020]33号);四川省中医药管理局项目(编号:2020JC0145)。

摘　　要：目的:通过体内实验研究和枢消积丸(Heshu Xiaoji prescription,HXP)对H22肝癌荷瘤小鼠肿瘤生成的影响以及对PI3K/AKT/mTOR信号通路相关蛋白的影响,探究其抑制肿瘤生长的作用机制。方法:制备H22肝癌荷瘤小鼠模型,随机分为空白对照组、模型组,和枢消积丸低、中、高剂量组(1.37、2.73、5.46g/kg),索拉非尼组(0.03g/kg),联合组(5.46g/kg和枢消积丸和0.03g/kg索拉非尼),每组5只,接种第6天开始给药,连续14d,记录小鼠体质量并观察一般情况;末次给药后次日处死小鼠,剥取肿瘤称质量,计算抑瘤率。苏木素-伊红(HE)染色检测观察肿瘤组织形态变化,酶联免疫吸附测定法(ELISA)检测肿瘤组织匀浆液IL-1β,TNF-α含量,免疫组化法以及Western-bloting法测定PI3K/AKT/mTOR信号通路中相关蛋白表达。结果:和枢消积丸能明显改善H22肝癌荷瘤小鼠的精神、活动状态,和枢消积丸低、中、高剂量组,索拉菲尼组,联合组的抑瘤率分别为23.9%、38.6%、64.5%、53.1%、76.6%(P<0.05);与模型组相比,各治疗组能明显降低肿瘤细胞密度,引起肿瘤细胞坏死;与模型组比较,各给药组肿瘤组织中IL-1β含量不同程度升高(P<0.05),与模型组比较,和枢消积丸高剂量组、索拉菲尼组以及联合组肿瘤组织中TNF-α含量不同程度升高(P<0.05);免疫组化结果显示,与模型组比较,联合组、和枢消积丸高剂量组PI3K,AKT,mTOR蛋白表达均明显下降;Western-bloting结果提示,与模型对照组相比较,各治疗组的P-PI3K、P-AKT、P-mTOR蛋白表达明显减少,和枢消积丸中、高剂量组、索拉菲尼组、联合组肿瘤组织的PPI3K/PI3K、P-AKT/AKT、P-mTOR/mTOR蛋白表达显著下调(P<0.05)。结论:和枢消积丸对H22肝癌荷瘤小鼠的肿瘤生成具有较为显著的抑制作用,其作用机制可能与下调PI3K/AKT/mTOR信号通路中关键蛋白表达有关。Purpose:To investigate the effect of Heshu Xiaoji prescription(HXP)on tumorigenesis in H22hepatocellular tumor-bearing mice as well as on PI3K/AKT/mTOR signaling pathway-related proteins through in vivo experiments and to explore the inhibitory tumor growth mechanism of action.Methods:H22hepatocellular tumor-bearing mice model was prepared,and the mice were randomly divided into blank control group,model group,HXP low,medium and high dose groups(1.37,2.73,5.46g/kg),sorafenib group(0.03g/kg),and combined group(5.46g/kg of HXP combined with 0.03g/kg of sorafenib).From the 6thday of inoculation,the mice were administered for 14consecutive days.The mice were weighed and the general conditions were observed.On the next day of the last administration,the mice were sacrificed,and the tumor was removed and weighed to calculate the anti-tumor rate.Hematoxylin-eosin(HE)staining was used to observe the morphological changes of the tumor tissues,enzyme-linked immunosorbent assay(ELISA)was used to detect the content of IL-1βand TNF-αin the homogenate of the tumor tissues,and immunohistochemistry and Western-blotting were used to detect the expression of proteins related to the PI3K/Akt/mTOR pathway.Results:HXP significantly improved the mental and activity status of H22hepatocellular tumor-bearing mice.The tumor inhibition rates of HXP low,medium and high dose groups,sorafenib group,and the combination group were 23.9%,38.6%,64.5%,53.1%,and 76.6%,respectively(P<0.05);compared with the model group,the therapeutic groups could significantly reduce the density of the tumor cells,cause tumor Compared with the model group,the content of IL-1βin the tumor tissues of each administration group was elevated to different degrees(P<0.05),and the content of TNF-αin HXP highdose group,the sorafenib group,and the combination group was elevated to different degrees(P<0.05).PI3K,Akt,mTOR protein expression were significantly decreased in the combined group and the HXP high-dose group;Western-blotting results suggested that P-PI3K,P-Akt,P

关 键 词：和枢消积丸 肝癌 中医药 PI3K/AKT/MTOR 

分 类 号：R285.5[医药卫生—中药学]