三叉苦总黄酮对裸鼠大肠癌移植瘤的抑制作用及机制  

Inhibitory effect and mechanism of total flavonoids from Melicope pteleifolia on transplanted tumor of colorectal cancer in nude mice

作　　者：刘义 安祯祥 何远利 高英 何秀义 LIU Yi;AN Zhenxiang;HE Yuanli;GAO Ying;HE Xiuyi(First Clinical Medical College,Guizhou University of Traditional Chinese Medicine,Guiyang 550001,China;Bijie Medical College,Guizhou Bijie 551700,China;Dept.of Gastroenterology,the First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine,Guiyang 550001,China)

机构地区：[1]贵州中医药大学第一临床医学院,贵阳550001 [2]毕节医学高等专科学校,贵州毕节551700 [3]贵州中医药大学第一附属医院消化内科,贵阳550001

出　　处：《中国药房》2024年第5期542-547,共6页China Pharmacy

基　　金：国家中医药管理局第五批全国中医临床优秀人才研修项目(No.国中医药人教函[2022]1号);贵州中医药大学中医脾胃病科技创新人才团队建设项目(No.贵中医TD合字[2022]005号);贵州中医药大学国家自然科学基金后补助项目(No.2019YFL171250206)。

摘　　要：目的研究三叉苦总黄酮对裸鼠大肠癌移植瘤生长的抑制作用及机制。方法采用裸鼠右侧腋窝皮下注射0.2 mL大肠癌细胞LoVo的方法构建大肠癌移植瘤模型。将造模成功的裸鼠分为模型组、5-氟尿嘧啶组(阳性对照,10 mg/kg)和三叉苦总黄酮高、低剂量组(25、12.5 mg/kg),另设不造模的正常组(含0.3%羧甲基纤维素钠的生理盐水),每组6只。各组裸鼠腹腔注射相应药液/溶剂,连续给药21 d。末次给药后,检测裸鼠移植瘤抑制率、肝/脾脏指数及血清中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)水平;观察裸鼠肿瘤组织病理形态学变化;采用免疫组化法检测裸鼠肿瘤组织中Toll样受体4(TLR4)、核因子κB亚基p65(NF-κB p65)蛋白表达水平;采用Western blot法检测裸鼠肿瘤组织中TLR4、髓样分化因子88(MyD88)、TNF受体相关蛋白6(TRAF6)、白细胞介素1受体关联激酶1(IRAK-1)、NF-κB p65、胱天蛋白酶3(caspase-3)蛋白表达水平。结果与模型组比较,三叉苦总黄酮高剂量组裸鼠瘤重(抑瘤率为36.91%)、肝/脾脏指数和血清中TNF-α、IL-6水平以及肿瘤组织中TLR4、MyD88、TRAF6、IRAK-1、NF-κB p65蛋白表达水平均显著降低(P<0.05或P<0.01),肿瘤组织中caspase-3蛋白表达水平显著升高(P<0.01);肿瘤细胞皱缩变形,核固缩、碎裂现象增多。结论三叉苦总黄酮可抑制裸鼠大肠癌移植瘤生长,其作用机制可能与减轻炎症反应,抑制TLR4/MyD88/NF-κB信号通路活性,促进大肠癌细胞凋亡有关。OBJECTIVE To study the inhibitory effect and mechanism of total flavonoids from Melicope pteleifolia(TF-MPL)on transplanted tumor of colorectal cancer in nude mice.METHODS The transplanted tumor model of colorectal cancer was induced by injecting 0.2 mL colorectal cancer cell LoVo subcutaneously via the right armpit of nude mice.After successful modeling,nude mice were randomly divided into model group,5-fluorouracil group(positive control,10 mg/kg),TF-MPL highdose and low-dose groups(25,12.5 mg/kg);a normal group(normal saline containing 0.3%carboxymethyl cellulose sodium)without modeling was additionally set up,with 6 mice in each group.Each group was intraperitoneally injected with the corresponding drug solution/solvent for 21 consecutive days.The inhibitory rate of the transplanted tumor,liver and spleen index,and the levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in serum were detected after the last medication;the morphological changes of tumor tissue were observed;immunohistochemical staining was used to detect protein expressions of Tolllike receptor 4(TLR4)and nuclear factor-κB subunit p65(NF-κB p65)in tumor tissue of nude mice.Western blot assay was used to detect protein expressions of TLR4,myeloid differentiation factor 88(MyD88),TNF receptor-associated factor 6(TRAF6),interleukin-1 receptor-associated kinase 1(IRAK-1),NF-κB p65 and caspase-3 in tumor tissue of nude mice.RESULTS Compared with the model group,TF-MPL high-dose group showed a significant decrease in tumor weight(inhibitory rate of 36.91%),liver and spleen index,serum levels of TNF-αand IL-6 and protein expressions of TLR4,MyD88,TRAF6,IRAK-1 and NF-κB p65(P<0.05 or P<0.01);the expression of caspase-3 protein was increased significantly(P<0.05),and more tumor cell shrinkage and deformation,nuclear pyknosis and fragmentation were observed.CONCLUSIONS TF-MPL can significantly inhibit the growth of transplanted tumor of colorectal cancer in nude mice,the mechanism of which may be associated with reducing inflammatory r

关 键 词：三叉苦总黄酮 大肠癌 炎症 Toll样受体4/髓样分化因子88/核因子κB信号通路 细胞凋亡 

分 类 号：R965[医药卫生—药理学] R285.5[医药卫生—药学]