黄地再生丸对再生障碍性贫血小鼠模型的影响及其机制探讨

Effect of Huangdi Zaisheng Pills on aplastic anemia mouse model and its mechanism

目的探讨黄地再生丸对再障小鼠模型的影响及其作用机制。方法将60只SPF级雄性小鼠,随机分为空白组(n=10)、造模组(n=50),射线造模。于第4、5、6天,对造模组小鼠行环磷酰胺25.0 mg/kg及氯霉素62.5mg/kg腹腔注射,空白组予以等体积生理盐水腹腔注射。第7天造模成功,将造模组小鼠随机均分为模型组,中药低、中、高剂量组,阳性药物组(n=10)。第8天灌胃给药。空白组、模型组蒸馏水10mL/kg灌胃。中药低、中、高剂量组分别予以相当于生药11.3g/kg、22.6g/kg、33.9 g/kg的丸剂灌胃。阳性药组灌胃18mg/kg环孢素A、18 mg/kg十一酸睾酮。连续21d。第7天,采血检测小鼠外周血白细胞(WBC)、红细胞(RBC)、血红蛋白(HGB)、血小板(PLT)、网织红细胞(Ret),评估造模情况。第22天,复检上述指标。髓细胞培养检测CFU-GM、CFU-E、BFU-E、CFU-Meg变化情况。第22天,取取材、固定、切片。检测CD34、血管红皮生长因子(VEGF)的表达。结果与模型组比较,中药组WBC、RBC、HGB、PLT均升高。与模型组比较,中药组Ret均升高。与模型组比较,中药组粒单核-巨噬系集落形成单位(CFU-GM)、红系集落形成单位(CFU-E)、红系爆式集落形成单位(BFU-E)、巨核系祖细胞(CFU-MEg)均有升高。与模型组比较,中药组微血管密度(MVD)均有明显升高。与模型组比较,中药组VEGF表达均升高。结论黄地再生丸通过增加骨髓中VEGF表达水平,促进血管生成,提高MVD,促使骨髓中CFU-GM、CFU-E、BFU-E、CFU-Meg升高,改善外周血象,起到改善再障的作用。

Objective To investigate the effect of Huangdi Zaisheng Pills on aplastic anemia mouse model and its mechanism.Methods Sixty SPF male mice were randomly divided into blank group(n=10),modeling group(n=50),and ray modeling.The mice in model group were intraperitoneally injected with cyclophosphamide 25.0mg/kg and chloramphenicol 62.5mg/kg,and blank group was intraperitoneally injected with an equal volume of normal saline on the 4th,5th and 6th days.On the 7th day,the model was successfully established,and the mice in model group were randomly divided into model group,TCM low-dose,medium-dose and high-dose groups,and positive drug group(n=10).Oral administration was given on the 8th day.Blank group and model group were given 10mL/kg of distilled water by gavage.TCM low-dose,medium-dose and highdose groups were administered with pills equivalent to 11.3g/kg,22.6g/kg,and 33.9g/kg of crude drug,respectively.Positive drug group was given 18mg/kg cyclosporine A and 18mg/kg testosterone undecanoate by gavage 21 consecutive days.On the 7th day,blood was collected to detect the white blood cell(WBC),red blood cell(RBC),hemoglobin(HGB),blood platelet(PLT),and reticulocyte(Ret)in peripheral blood of mice to evaluate the modeling situation.On the 22nd day,recheck the above indicators,myeloid cell culture was used to detect the changes of CFU-GM,CFU-E,BFU-E and CFU-Meg,materials were collected,fixed and sliced.The expressions of CD34 and vascular endothelial growth factor(VEGF)were detected.Results Compared with model group,WBC,RBC,HGB and PLT in TCM low,medium and high dose groups were all increased;compared with model group,CFU-GM,CFU-E,BFU-E and CFU-Meg in TCM low,medium and high dose groups were all increased;compared with model group,the microvessel density(MVD)in TCMlow,medium and high dose groups was significantly increased;compared with model group,the expression of VEGF in TCMlow,medium and high dose groups increased.Conclusion Huangdi Zaisheng Pills increase the expression level of VEGF in the bone marrow,promotes angiogenesis,increases MVD,promotes the increase of CFU-GM,CFU-E,BFU-E and CFU-Meg in the bone marrow,improves peripheral blood,and plays a role in improving aplastic anemia.