乙酰苯肼致血虚证模型小鼠的证候物质基础初探  

Study of Material Base on Acetylphenylhydrazine Induced Blood Deficiency Syndrome Model Mice

作　　者：钱宏梁 潘志强[1] 王晓敏[1] 方肇勤[1] QIAN Hong-liang;PAN Zhi-qiang;WANG Xiao-min;FANG Zhao-qin(Basic Medical School,Shanghai University of Traditional Chinese Medicine,Shanghai (201203))

机构地区：[1]上海中医药大学基础医学院,上海201203

出　　处：《中国中西医结合杂志》2018年第12期1490-1495,共6页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金面上项目(No.81473562)

摘　　要：目的研究乙酰苯肼引起的血虚证模型小鼠证候可能的物质基础。方法以雄性ICR小鼠为对象,采用乙酰苯肼170 mg/kg皮下注射,第1天和第4天各1次制作血虚证模型,并设立正常对照组。动态观察小鼠体重变化,显微镜下观察血液细胞与股骨骨髓细胞,取肾脏、肾上腺和骨髓分别抽提RNA,以实时荧光定量PCR技术检测相关基因表达、以ELISA方法检测血清皮质酮含量。结果与正常对照组比较,乙酰苯肼小鼠造模3天后体重显著下降(P<0.05),小鼠呈现虚弱征象,小鼠脾脏代偿性增大而胸腺却明显萎缩(P<0.01)。乙酰苯肼组小鼠血液红细胞形态异常、正常红细胞数量减少、白细胞代偿性增多;骨髓成熟的环状核细胞减少。与正常对照组比较,乙酰苯肼组小鼠骨髓IL-5和IL-6基因表达显著升高(P<0.05),而巨噬细胞集落刺激因子(M-CSF)、IL-1b和IL-7基因表达显著下降(P<0.05);肾脏促红细胞生成素(Epo)、粒细胞集落刺激因子(G-CSF)和M-CSF基因表达也显著升高(P<0.05,P<0.01),而粒细胞-巨噬细胞集落刺激因子(GM-CSF)基因表达则被乙酰苯肼抑制(P<0.05);肾上腺Cyp21a1与Cyp11b1基因表达显著升高(P<0.05);小鼠血清皮质酮含量明显升高(P<0.01)。结论乙酰苯肼所复制的证候以血虚证为主,脾脏肿大、肾脏Epo基因代偿性高表达以及继发肾上腺皮质功能虚性亢奋是该证候物质与功能变化最突出特征。Objective To study the material base on acetylphenylhydrazine induced blood deficiency syndrome(BDS) model mice. Methods Male ICR mice were subcutaneously injected with acetylphenylhydrazine(170 mg/kg). BDS model was prepared on the 1st day and the 4th day. A normal control group was also set up by injecting normal saline at the same dosage. Body weight was dynamically observed. Changes of each organ were detected after sacrificed. Blood and femur bone marrow smears were made and observed under a microscope. RNAs were extracted from kidney, adrenal gland, and bone marrow using TRIzol kit. Related gene expressions were detected by real-time quantitative PCR, and serum corticosterone content was tested by ELISA. Results Compared with the normal control group, body weight of mice in the acetylphenylhydrazine group significantly decreased after 3 days modeling(P<0.05). The mice showed weak signs. Their spleens were compensatively enlarged, but the thymus was severely atrophic(P<0.01). Their erythrocytes′ morphologies were abnormal and the numbers of normal red blood cells were reduced, and leucocyte were compensatively increased. Mature ring neutrophils in bone marrow were reduced. Compared with the normal control group, gene expressions of bone marrow IL-5 and IL-6 increased significantly(P<0.05), while gene expressions of M-CSF, IL-1b, and IL-7 decreased significantly(P<0.05). Gene expressions of renal Epo, G-CSF, and M-CSF significantly increased(P<0.05, P<0.01), but the gene expression of renal GM-CSF were inhibited by acetylphenylhydrazine(P<0.05). Gene expressions of adrenal gland Cyp21a1 and Cyp11b1 increased significantly(P<0.05). Moreover, serum corticosterone content increased obviously in the acetylphenylhydrazine group(P<0.01). Conclusions Acetylphenylhydrazine could cause typical BDS. The most prominent features included splenomegaly, compensatory high expression of renal Epo gene, and secondary adrenal hyperactivity.

关 键 词：乙酰苯肼 血虚证 证候 促红细胞生成素 环状核细胞 

分 类 号：R-332[医药卫生] R255.7