补髓生血丸对再生障碍性贫血模型小鼠骨髓VEGF及 Notch-1蛋白表达的影响  

Effect of Busui Shengxue Pills (补髓生血丸) on VEGF and Notch-1 Protein Expression in Bone Marrow of Mice with Aplastic Anemia

作　　者：杨志勇[1] 邓梦云 李莉[2] 童小慧 王雅娟[2] 王桐生[2,3] YANG Zhiyong;DENG Mengyun;LI Li;TONG Xiaohui;WANG Yajuan;WANG Tongsheng(Department of General Medicine,Fuyang People's Hospital,Fuyang 236003;Department of Pharmacology,Anhui University of Chinese Medicine,Hefei 230012;Anhui Province Key Laboratory of Chinese Medicinal Formula,Hefei 230012)

机构地区：[1]阜阳市人民医院全科医学科,阜阳236003 [2]安徽中医药大学药理学教研室,合肥230012 [3]中药复方安徽省重点实验室,合肥230012

出　　处：《中药药理与临床》2023年第9期14-18,共5页Pharmacology and Clinics of Chinese Materia Medica

基　　金：安徽中医药大学校企联合项目(编号:RD18200160)。

摘　　要：目的:研究补髓生血丸对再生障碍性贫血模型小鼠的改善作用,并从调控骨髓造血微环境角度探讨其作用机制。方法:取昆明种雄性小鼠,随机分为正常对照组、模型对照组、补髓生血丸2.3、4.5、9 g/kg组和再造生血片2.7 g/kg组,每组15只。除正常对照组外,其余各组采用白消安10 mg/kg和环磷酰胺20 mg/kg交替腹腔注射给药,连续12 d,制备再生障碍性贫血小鼠模型。造模同时各组小鼠按剂量灌胃给药,连续30 d。末次给药禁食不禁水12 h,颈椎脱臼法处死小鼠,分离双侧股骨,采用瑞士-姬姆萨法观察小鼠骨髓涂片;HE法观察骨髓病理情况;采用ELISA法检测小鼠骨髓上清中血管内皮生长因子(VEGF)、血小板生成素(TPO)、血管细胞粘附因子-1(VCAM-1)的含量;采用免疫组化法检测小鼠骨髓微血管密度(MVD)、小鼠血管VEGF和Notch-1的蛋白表达。结果:与正常对照组比较,模型对照组小鼠外周血红细胞(RBC)、白细胞(WBC)、血小板(PLT)数量、血红蛋白(HGB)含量明显降低(P<0.05);骨髓腔空虚,大量脂肪细胞代替造血细胞,未见巨核细胞,骨髓上清中VCAM-1含量明显降低(P<0.05);骨髓MVD显著降低(P<0.01),VEGF、Notch-1蛋白表达明显下调(P<0.05);连续给药30 d后,与模型对照组比较,给药各组小鼠外周血象指标均有不同程度的升高,骨髓造血功能改善明显,骨髓可见增生现象,有核细胞逐渐增多,巨核细胞多见;再造生血片2.7 g/kg组及补髓生血丸各组骨髓上清中VCAM-1含量显著升高(P<0.01);骨髓MVD显著增加(P<0.01);除补髓生血丸2.3 g/kg组外,其余各组骨髓VEGF和Notch-1蛋白表达显著上调(P<0.01)。结论:补髓生血丸对再生障碍性模型小鼠有一定防治作用,其机制可能为通过增加骨髓MVD,上调VEGF和Notch1蛋白表达,促进血管新生,改善骨髓造血微环境。Objective:To investigate the therapeutic effects of Busui Shengxue Pills(补髓生血丸)on a mouse model of aplastic anemia and explore its mechanism of action from the perspective of regulating the bone marrow hematopoietic microenvironment.Methods:Male Kunming mice were randomly divided into a normal control group,a model control group,Busui Shengxue Pills(2.3,4.5,and 9 g/kg)groups,and a Zaizao Shengxue(再造生血)Tablets(2.7 g/kg)group,with 15 mice in each group.Except for the normal control group,all other groups were injected intraperitoneally with busulfan(10 mg/kg)and cyclophosphamide(20 mg/kg)alternately for 12 consecutive days to establish the mouse model of aplastic anemia.Concurrently,the mice in each group were orally administered the corresponding doses of Busui Shengxue Pills or Zaizao Shengxue Tablets for 30 consecutive days.After a 12-hour fasting period with free access to water following the last administration,the mice were euthanized by cervical dislocation.Bilateral femurs were isolated for Weigert's-Giemsa staining to observe bone marrow smears.HE staining was performed to examine bone marrow pathology.ELISA was used to measure the levels of vascular endothelial growth factor(VEGF),thrombopoietin(TPO),and vascular cell adhesion molecule-1(VCAM-1)in the mouse bone marrow supernatant.Immunohistochemistry was conducted to evaluate the microvascular density(MVD)and protein expression of VEGF,VCAM-1,and Notch-1 in the bone marrow.Results:Compared with the normal control group,the model control group showed significantly decreased levels of red blood cells(RBC),white blood cells(WBC),hemoglobin(HGB),and platelets(PLT)in peripheral blood(P<0.05).The bone marrow exhibited empty cavities,a large number of fat cells replacing hematopoietic cells,and absence of megakaryocytes.The levels of VEGF and VCAM-1 in the bone marrow supernatant were reduced,with VCAM-1 showing a significant decrease(P<0.05).MVD in the bone marrow significantly decreased(P<0.01),and the protein expression of VEGF and Notch

关 键 词：补髓生血丸 再生障碍性贫血 微血管密度 血管内皮生长因子 血管细胞粘附因子-1 NOTCH-1 

分 类 号：R285.5[医药卫生—中药学]