补肾益髓生血法对^(60)Co-γ联合环磷酰胺诱导再生障碍性贫血大鼠骨髓造血及免疫功能的影响

An experimental study of immune and hematopoietic effects of Bushen Yisui Shengxue treatment menthods on aplastic anemia rats induced by ^(60)Co-γ and CTX

目的:探讨补肾益髓生血法对60C o-γ射线联合环磷酰胺(CTX)诱导再生障碍性贫血(AA)大鼠骨髓造血及免疫功能的影响。方法:110只S D大鼠,随机分为正常组10只,造模组100只。造模组大鼠用4.0Gy 60Co-γ射线一次性照射后,第4天腹腔注射CXT 25.0mg/kg,连续3d,造模成功后,存活大鼠随机分为模型组、司坦唑醇组、益髓生血组、温肾生血组和滋肾生血组。用药2周后,处死大鼠,经股动脉取血检测白细胞(WBC)、红细胞(RBC)、血小板(PLT)及血红蛋白(HGB),流式细胞术检测CD3、CD4、CD8,血涂片瑞氏染色观察血细胞形态,骨髓涂片观察骨髓细胞形态,骨髓悬液检测骨髓有核细胞数量。结果:外周血象,与模型组比较,滋肾生血组WBC、RBC、PLT及HGB均明显升高(P<0.05,P<0.01);血涂片,滋肾生血组细胞通透性好转,RBC和WBC形态趋于正常,有核细胞增多,退化细胞减少;骨髓涂片,滋肾生血组,细胞脂肪滴减少,有核细胞增多,非造血细胞减少;骨髓有核细胞,与模型组比较,滋肾生血组显著增加(P<0.01);流式细胞术,CD8、CD3滋肾生血组明显减低(P<0.05);CD4、CD4/CD8滋肾生血组明显升高(P<0.05,P<0.01)。结论:补肾益髓生血法可能通过调控AA免疫功能,起到显著的治疗作用;对比研究显示,滋肾生血法明显优于益髓生血法和温肾生血法,这对于阐释再障肾藏精生血理论具有重要意义。

Objective： To observe the influence of Bushen Yisui Shengxue treatment methods on immune and hematopoietic functions in aplastic anemia （AA） rats induced by 60Co-γ and cyclophosphamide （CXT）. Methods： 110 male SD rats were randomly divided into a normal group （10 rats） and experimental group （100 rats）. Rats in the experimental group were injected with 4.0Gy 60Co- γ Irradiate, then with CTX （25.0mg/kg） for the next 3 days. Once the AA models were successfully developed, those rats were randomly divided into the model group, stanozolol group, yisui group, wenshen group and zishen group according to weights. All rats were killed after two weeks of treatment, and blood samples from their femoral artery were collected and detected CD3, CD4, and CD8 were tested with flow cytometry. Wright stains were used on blood and bone marrow smears to observe blood cell and marrow cell morphology. The number of bone marrow cells were obtained by detecting the bone marrow suspension. Results： RBC, WBC, HGB, and PLT counts were significantly lower （P〈0.01） in the peripheral blood of rats from the model group than the normal group, while there were signficantly increased in the treatment groups （P〈0.05, P〈0.01）. Compared against the treatment groups, for blood smears in the model group, Poor cell permeability, white blood ceils were fewer in number, and cell degradation was more common; for bone marrow smears of the model group significantly more fat droplets, reduced nuclear proliferation, fewer erythrocytes, myelocytes, and megakaryocytes counted, and the appearance of non-hematopoietic cells were observed. The number of nucleated cells were significantly greater in treatment groups than in the bone marrow of the model group （P〈0.01）. For flow cytometry results, compared with the normal group, levels of CD8 and CD3 were significantly increased in the model group （P〈0.05） but significantly lowered in the treatment groups （P〈0.05）. Conversely, CD4 was significantly decreased in the model group （P〈0.01） and signficantly increased in the treatment groups （P〈0.05）. The CD4/CD8 ratio was significantly reduced in the model group （P〈0.05）, while it was significantly increased in the treatment groups （P〈0.05, P〈0.01）. Conclusion： Through regulation of AA immune function, BushenYisui Shengxue method may reduce the pathological damage to the bone marrow increase peripheral blood cells, t and thus play a significant role in the treatment of AA. Zishen Shengxue method is better than Wenshen Shengxue method. These results may provide the physiological basis to better understanding the theory behind the kidney＇s role in life-essence storage and blood production.