九味阿胶膏对血虚证小鼠的补血作用及其机制研究

Blood Enriching Effect and Mechanism of Jiuwei Ejiao Ointment on Mouse Model of Blood Deficiency Syndrome

目的研究九味阿胶膏对血虚证小鼠的补血作用及其作用机制。方法采用乙酰苯肼(APH)和环磷酰胺(CTX)联合应用建立血虚证小鼠模型。ICR小鼠60只,随机分为正常组,模型组,四物颗粒组,九味阿胶膏高、中、低(15. 6、7. 8、3. 9g·kg·kg^(-1))组,每组10只。连续给药14 d后,取小鼠血清、骨髓组织。血液分析仪测定小鼠外周血中白细胞(WBC)、红细胞(RBC)、血红蛋白(HGB)水平; HE染色观察骨髓组织病理学改变; ELISA法测定小鼠血清中促红细胞生成素(EPO)、白细胞介素-3(IL-3)、肿瘤坏死因子(TNF)-α、中粒细胞集落刺激因子(G-CSF)、巨噬细胞集落刺激因子(GM-CSF)水平; Westernblot法测定小鼠肾脏组织中EPO、骨髓组织中G-CSF、GM-CSF蛋白表达水平。结果与模型组比较,九味阿胶膏高、中、低剂量可明显升高外周血WBC含量,高、中剂量可明显升高外周血RBC、HGB含量;高、中剂量组骨髓增生程度均有明显好转,三系明显增生,非造血组织及脂肪细胞明显减少,造血组织结构明显修复;高、中、低剂量可明显降低血清中TNF-α含量;高、中剂量可明显升高血清中EPO、IL-3、G-CSF、GM-CSF含量,明显增加肾脏组织中EPO和骨髓组织中G-CSF、GM-CSF蛋白表达水平。结论九味阿胶膏对血虚证小鼠具有明显的补血作用,其机制可能与其调控促造血生长因子的分泌,减轻负向造血因子对造血功能的抑制有关。

OBJECTIVE To study the blood enriching effect and mechanism of Jiuweiejiao ointment on mouse model of blood deficiency syndrome.METHODS Blood deficiency syndrome model was established with cyclophosphamide(CTX) and acetylphenylhydrazine(APH). Total of 60 ICR mice were divided into normal group, model group, Siwu granule group, Jiuweiejiao ointment high-dose, medium-dose and low-dose group (15.6, 7.8, 3.9 g·kg^-1). Each group mouse were treated with the respective drug through intragastrical administration once a day for consecutive 14 d. The levels of red blood cells (RBC), leukocytes (WBC), hemoglobin (HGB) level in blood of mice were detected by blood analyzer; the histopathological changes of bone marrow were determined by HE staining; the levels of erythropoietin (EPO), interleukin-3 (IL-3), tumor necrosis factor-α (TNF-α), granulocyte colony stimulating factor (G-CSF) and granulocyte-macrophage colony stimulating factor (GM-CSF) in serum were measured by enzyme linked immunosorbent assay (ELISA) method; the expression level of EPO protein in kidney tissue, G-CSF and GM-CSF protein in bone marrow tissue were detected by Western-blot method.RESULTS Compared with model group, Jiuweiejiao ointment could significantly increase the levels of RBC, WBC and HGB in blood of mice, markedly improve bone marrow hyperplasia, reduce the non-hematopoietic tissue and adipocytes, obviously repaire the hematopoietic tissue structure; significantly decrease the level of TNF-α and increase the level of EPO, IL-3, G-CSF and GM-CSF in serum. In addition, Jiuweiejiao ointment could also obviously increase the expression of EPO protein in kidney tissue and G-CSF, GM-CSF protein in bone marrow tissue compared with that in model group.CONCLUSION Jiuweiejiao ointment has obvious blood enriching effects, which may be related with promoting the secretion of hematopoietic growth factors and reducing the inhibition of hematopoietic function by negative hematopoietic factor.