益髓方改善胃癌小鼠化疗后骨髓抑制机制研究  

Study on Inhibition Mechanism of Yisui Recipe on Improving Bone Marrow Suppression in Mice with Gastric Cancer after Chemotherapy

作　　者：黄春风 杨先玉[1] 郑慧华[1] 刘大毛 陈盼敏 喻敏 王征 HUANG Chunfeng;YANG Xianyu;ZHENG Huihua;LIU Damao;CHEN Panmin;YU Min;WANG Zheng(The Affiliated Hospital of Jiangxi University of Chinese Medicine,Yichun Jiangxi China 336000;The Second Affiliated Hospital of Jiangxi University of Chinese Medicine,Yichun Jiangxi China 336000;Shangrao People's Hospital,Shangrao Jiangxi China 334000)

机构地区：[1]江西中医药大学附属医院,江西宜春330000 [2]江西中医药大学第二附属医院,江西南昌336000 [3]上饶市人民医院,江西上饶334000

出　　处：《中医学报》2021年第12期2627-2635,共9页Acta Chinese Medicine

基　　金：江西省科技计划项目(2015ZBBG70017)。

摘　　要：目的:观察益髓方对胃癌小鼠化疗后骨髓抑制的改善作用及对骨髓造血功能的促进作用,并初步探究其作用机制。方法:将50只BALB/c小鼠随机分为正常组、模型组、吉粒芬组(12μg·kg^(-1))、益髓方组(40.4 g·kg^(-1))及吉粒芬+益髓方组(12μg·kg^(-1)吉粒芬+40.4 g·kg^(-1)益髓方)。除正常组外,其余组小鼠皮下接种小鼠胃癌细胞(mouse forestomach carcinoma,MFC)建立胃癌荷瘤小鼠模型,当瘤体积达到200 mm^(3)且无破溃时,腹腔注射环磷酰胺(cyclophosphamide,CTX)建立胃癌荷瘤小鼠化疗后骨髓抑制模型。造模完成第2天开始给予相应药物干预,正常组和模型组小鼠给予等体积生理盐水,每天1次,连续7 d。全自动血细胞分析仪检测小鼠血常规指标;流式细胞仪检测外周血免疫细胞亚群比例及骨髓细胞周期变化;ELISA法检测血清白细胞介素-3(interleukin-3,IL-3)、IL-6、血小板生成素(thrombopoietin,TPO)、促红细胞生成素(erythropoietin,EPO)和粒细胞-巨噬细胞集落刺激因子(granulocyte macrophage colony stimulating factor,GM-CSF)的水平;瑞氏-吉姆萨染色检测骨髓有核细胞(bone marrow nucleated cells,BMNCs)数量;Western Blot检测小鼠骨髓细胞中Jag1、Notch2、Numb1及Numb2的蛋白表达水平。结果:与正常组比较,模型组小鼠外周血白细胞(white blood cell,WBC)、红细胞(red blood cell,RBC)、血小板(platelet,PLT)及BMNCs计数显著降低(P<0.05),血红蛋白(hemoglobin,Hb)总量显著下降(P<0.05),CD_(3)^(+)、CD_(4)^(+)、CD_(8)^(+)细胞的百分比及CD_(4)^(+)/CD_(8)^(+)、S期细胞比例显著降低(P<0.05),血清IL-3、IL-6、TPO、EPO、GM-CSF的水平和骨髓细胞Numb1、Numb2的蛋白表达水平均显著降低(P<0.05),G0/G1期细胞比例及Jag1、Notch2的蛋白表达水平显著增加(P<0.05);与模型组比较,益髓方组小鼠外周血WBC、PLT、BMNCs计数显著升高(P<0.05),CD_(3)^(+)、CD_(4)^(+)、CD_(8)^(+)细胞的百分比及CD_(4)^(+)/CD_(8)^(+)、S期及G2/M�Objective:To observe the effect of Yisui recipe on improving bone marrow suppression and promoting bone marrow hematopoietic function in gastric cancer mice after chemotherapy,and to explore its mechanism of action.Methods:Fifty BALB/c mice were randomly divided into normal group,model group,Jilifen group(12 μg·kg^(-1)),Yisui recipe group(40.4 g·kg^(-1)),and JiLifen+Yisui recipe group(12 μg·kg^(-1) Ji Lifen+40.4 g·kg^(-1) Yisui Fang).Except for the normal group,the mice in other groups were subcutaneously inoculated with MFC cells to establish a gastric cancer tumor-bearing model.When the tumor volume reached 200 mm^(3) and there was no ulceration,the bone marrow suppression model of gastric cancer-bearing mice after chemotherapy was established by intraperitoneal injection of cyclophosphamide(CTX).After the completion of the model building,the corresponding drug intervention was started on the second day.The mice in the normal group and the model group were given the same amount of normal saline by gavage once a day for 7 days.After the experiment,the automatic blood cell analyzer was used to detect the blood routine of mice.The flow cytometer was used to detect the proportion of peripheral blood immune cell subsets and the changes in the bone marrow cell cycle.The levels of serum interleukin-3(IL-3),IL-6,thrombopoietin(TPO),erythropoietin(EPO),and granulocyte-macrophage colony-stimulating factor(GM-CSF) were detected by ELISA.The number of nucleated cells in bone marrow was detected by Wright Giemsa staining.The protein expression levels of Jag1,Notch2,numb1 and numb2 in mouse bone marrow cells were detected by Western blot.Results:Compared with the normal group,the white blood cell(WBC),red blood cell(RBC),platelet(PLT),and BMNCs counts in the model group were significantly reduced(P<0.05).The total amount of hemoglobin(hemoglobin,Hb) decreased significantly(P<0.05).The percentage of CD_(3)^(+) cells,CD_(4)^(+) cells,and CD_(8)^(+) cells and the ratio of CD_(4)^(+)/CD_(8)^(+) and S-phase cells were signi

关 键 词：益髓方 胃癌 骨髓抑制 化疗 NOTCH信号通路 小鼠 

分 类 号：R285.5[医药卫生—中药学]