益肾祛痛颗粒灌服对小鼠乳腺癌骨转移灶的改善作用及其机制  

Inhibitory effect and mechanism of Yishen Qutong granules on bone metastasis of breast cancer in mice

作　　者：殷玉琨 陈佳阳 常金圆 刘丽星 江正龙 冯利 YIN Yukun;CHEN Jiayang;CHANG Jinyuan;LIU Lixing;JIANG Zhenglong;FENG Li(Traditional Chinese Medicine Department,National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100021,China)

机构地区：[1]国家癌症中心,国家肿瘤临床医学研究中心,中国医学科学院北京协和医学院肿瘤医院中医科,北京100021

出　　处：《山东医药》2024年第23期52-56,共5页Shandong Medical Journal

基　　金：国家自然科学基金委员会项目(81904215);中国医学科学院“中国健康长寿创新大赛”项目(2020-JKCS-001);中国医学科学院中央级公益性科研院所基本科研业务费专项(2019XK320072);北京市中医药科技发展资金项目(69JJ-2023-82)。

摘　　要：目的观察益肾祛痛颗粒灌服对小鼠乳腺癌骨转移灶的改善作用,并探讨其可能的机制。方法将84只雌性BABL/c小鼠随机分为7组,每组12只。其中模型组、ML-385组及低、中、高剂量组均采用左侧胫骨种植小鼠亲骨性乳腺癌高转移细胞系4T1.2细胞悬液的方法建立乳腺癌骨转移模型,种植后第1天低、中、高剂量组分别给予益肾祛痛颗粒水溶液2.275、4.55、9.1 g/kg灌服并腹腔注射生理盐水,ML-385组给予蒸馏水灌服并腹腔注射溶于生理盐水的细胞核因子E2相关因子2(Nrf2)抑制剂ML-38520 mg/kg,空白组、假手术组、模型组给予等体积蒸馏水灌服并腹腔注射生理盐水,1次/天,连续给药28 d。记录各组小鼠分组处理1~4周的体质量,连续给药28 d后处死,称取肿瘤质量。取各组小鼠左侧胫骨,micro-CT扫描后分析骨体积分数(BV/TV)、骨小梁厚度(Tb.Th)、骨小梁数目(Tb.N)及骨小梁间隙(Tb.Sp),HE染色后观察组织病理情况,酒石酸酸性磷酸酶染色后分析破骨细胞数量及破骨细胞与骨组织接触面长度,免疫组化法检测Kelch样环氧氯丙烷相关蛋白1(Keap1)、Nrf2蛋白阳性表达率。结果分组处理1~4周,各组小鼠体质量呈持续上升趋势;分组处理第4周,模型组体质量低于其余各组(P均<0.05)。与模型组比较,各组肿瘤质量均降低,但仅ML-385组和中剂量组P均<0.05。空白组和假手术组左侧胫骨组织病理学观察均正常,模型组左侧胫骨有明显骨转移瘤形成、细胞成分以成骨性骨转移为主,ML-385组、中剂量组左侧胫骨骨转移瘤形成及骨质破坏程度均较模型组减轻,低、高剂量组与模型组相近。与空白组及假手术组比较,模型组左侧胫骨组织BV/TV、Tb.N、Tb.Sp、Tb.Th、破骨细胞数量、破骨细胞接触面长度及Keap1、Nrf2阳性表达率均升高(P均<0.05);与模型组比较,ML-385组左侧胫骨组织BV/TV、Tb.N、Tb.Sp、Tb.Th及Keap1、Nrf2阳性表达率均降低,中剂�Objective To observe the inhibitory effect of intragastric administration of Yishen Qutong granules(YSQTG)on breast cancer bone metastasis in mice,and to explore its possible mechanism of action.Methods Eightyfour female BABL/c mice were randomly divided into seven groups,with 12 mice in each group.Mice in the model group,ML-385 group,and low,medium,and high dose groups were all implanted with the bone-affine breast cancer high meta⁃static cell line 4T1.2 cell suspension in the left tibia to create the breast cancer bone metastasis models.On the first day af⁃ter implantation,mice in the low,medium,and high dose groups were given YSQTG aqueous solution at 2.275,4.55,and 9.1 g/kg,respectively,by gavage,followed by intraperitoneal injection of normal saline.Mice in the ML-385 group were given distilled water by gavage and intraperitoneal injection of nuclear factor E2-related factor 2(Nrf2)inhibitor ML-385(20 mg/kg)dissolved in normal saline.Mice in the blank group,sham operation group,and model group were given equal volume of distilled water by gavage and intraperitoneal injection of normal saline.The dose was administered once a day,for 28 consecutive days.The body weight of mice in each group was recorded from 1 to 4 weeks after treatment,and after 28 days of continuous administration,the mice were sacrificed and the tumor mass was weighed.The left tibia of mice in each group was harvested,and after micro-CT scanning,the bone volume fraction(BV/TV),trabecular thickness(Tb.Th),trabecular number(Tb.N),and trabecular space(Tb.Sp)of cancellous and cortical bone were analyzed.Histopath⁃ological conditions were observed after HE staining,and the number of osteoclasts and the length of contact surface between osteoclasts and bone tissues were analyzed after tartrate acid phosphatase staining.Immunohistochemical method was used to detect the positive expression rates of Kelch-like ECH-associated protein 1(Keap1)and Nrf2 proteins.Results During the 1 to 4 weeks of treatment,the body weight of mice in each group sho

关 键 词：乳腺癌 骨转移 益肾祛痛颗粒 破骨细胞 Kelch样环氧氯丙烷相关蛋白1 细胞核因子E2相关因子2 

分 类 号：R273[医药卫生—中西医结合]