靶向表皮生长因子受体的百秋李醇纳米颗粒抑制非小细胞肺癌裸鼠皮下移植瘤生长的作用及机制  

Inhibitory effect and mechanism of epidermal growth factor receptor⁃targeted nanoparticles loaded with patchouli alcohol on growth of subcutaneous transplantation tumor of non⁃small cell lung carcinoma growth in nude mice

作　　者：段振东 张欣欣 陈红宇[2] 杨柳[2] DUAN Zhendong;ZHANG Xinxin;CHEN Hongyu;YANG Liu(Zhejiang Zhoushan Hospital of Traditional Chinese Medicine,Zhoushan,Zhejiang 316000,China;Baoshan Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201900,ChinaAbstract)

机构地区：[1]浙江省舟山市中医院,浙江舟山316000 [2]上海中医药大学附属宝山医院,上海201900

出　　处：《上海中医药杂志》2024年第6期65-72,共8页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(82274338);上海市科委项目(21Y11922200)。

摘　　要：目的探讨靶向表皮生长因子受体(EGFR)的百秋李醇(PA)纳米颗粒对非小细胞肺癌(NSCLC)肿瘤生长的抑制作用及其机制。方法①利用二硬脂酰基磷脂酰乙醇胺-聚乙二醇-EGFR靶向肽AA1(DSPE-PEG-AA1)脂质体负载PA,构建靶向EGFR的PA脂质体纳米颗粒(EGFR-PA-NP)。②将人NSCLC A549细胞(2×106个/只)接种于裸鼠腋下,建立肺癌异种皮下移植瘤裸鼠模型。10只模型裸鼠随机分为PA脂质体纳米颗粒(PA-NP)组和EGFR-PA-NP组(n=5),分别尾静脉注射给予20 mg·kg^(-1)相应药物,24 h后剥取肿瘤、癌旁、肝、肺、心、脑、肾、脾、胃、小肠等组织,液相色谱-质谱(LC-MS)法检测药物在体内的组织分布情况。③20只模型裸鼠随机分为模型组(100μL)、PA组(15 mg·kg^(-1))、PA-NP组(15 mg·kg^(-1))和EGFR-PA-NP组(15 mg·kg^(-1))(n=5),分别尾静脉注射给予相应干预,隔日1次,共28 d。28 d后观察药物对裸鼠皮下移植瘤生长的影响。通过原位末端转移酶标记(TUNEL)检测法检测皮下移植瘤中肿瘤细胞凋亡情况,免疫荧光染色法检测皮下移植瘤中血小板-内皮细胞黏附分子(CD31)表达情况,免疫组化染色法和Western blot法检测皮下移植瘤中蛋白激酶B(Akt)、磷酸化(p)-Akt、哺乳动物雷帕霉素靶蛋白(mTOR)和p-mTOR表达水平。结果①制备的EGFR-PA-NP粒径为(122±6.90)nm、Zeta电位为(-24.28±3.76)mV、包封率为85.24%、载药量为8.09%,符合纳米颗粒特征。②LC-MS法检测药物组织分布结果显示,EGFR-PA-NP组药物在裸鼠瘤体内的富集量是PA-NP组的3倍,显著高于PA-NP组(P<0.05)。③在裸鼠A549皮下移植瘤模型中,PA组、PA-NP组和EGFR-PA-NP组裸鼠瘤体体积明显小于模型组(P<0.05),而EGFR-PA-NP组瘤体体积明显小于PA-NP组(P<0.05)。与PA-NP组相比,EGFR-PA-NP组肿瘤中凋亡细胞数量显著增加(P<0.05),CD31蛋白表达水平显著降低(P<0.05),并且p-Akt、p-mTOR表达水平显著降低(P<0.05)。结论EGFR-PA-NP能够通过促进肿瘤细�Objective To investigate the inhibitory effect of epidermal growth factor receptor(EGFR)-targeted nanoparticles loaded with patchouli alcohol(PA)on the growth of non-small cell lung carcinoma(NSCLC)and its mechanism.Methods①Distearylphosphatidylethanolamine-polyethylene glycol-EGFR targeting peptide AA1(DSPE-PEG-AA1)liposomes were used to load PA,to construct EGFR-targeted PA liposome nanoparticles(EGFR-PA-NP).②A subcutaneous xenograft model of lung cancer in nude mice was established by inoculating human NSCLC A549 cells(2×106 cells/mouse)into the armpit of nude mice.Ten model nude mice were randomly divided into PA liposome nanoparticles(PA-NP)group and EGFR-PA-NP group(n=5),and the corresponding drugs were given by tail vein injection at a dose of 20 mg·kg^(-1).After 24 h,the tumor,adjacent tissues,liver,lung,heart,brain,kidney,spleen,stomach,small intestine were colleted.Liquid chromatography-mass spectrometry(LC-MS)method was used to detect the drugs distribution in vivo.③Twenty model nude mice were randomly divided into model group(100μL),PA group(15 mg·kg^(-1)),PA-NP group(15 mg·kg^(-1))and EGFR-PA-NP group(15 mg·kg^(-1)),and corresponding interventions were given by tail vein injection,once every other day,for 28 d.After 28 d,the effect of drugs on the growth of subcutaneous transplanted tumor in nude mice was observed.The apoptosis of tumor cells in subcutaneously transplanted tumors was detected by terminal-deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling(TUNEL)assay.The expression of platelet endothelial cell adhesion molecule-1(CD31)was detected by immunofluorescence staining.The expression levels of protein kinase B(Akt),phosphorylated(p)-Akt,mammalian target of rapamycin(mTOR)and p-mTOR were detected by immunohistochemical staining and Western blot.Results①The prepared EGFR-PA-NP had a particle size of(122±6.90)nm,Zeta potential of(-24.28±3.76)mV,encapsulation efficiency of 85.24%,and drug loading capacity of 8.09%,which were consistent with the characteristics of

关 键 词：肺癌 百秋李醇 木防己 纳米颗粒 药物载体 抗肿瘤 中药研究 

分 类 号：R285.5[医药卫生—中药学]