健脾补肾方对再生障碍性贫血大鼠骨髓造血功能的影响及机制研究

Effects of Jianshenbupi prescription on hematopoietic function in rats with aplastic anemia and related action mechanisms

目的观察健脾补肾方对再生障碍性贫血(AA)大鼠骨髓造血功能的影响,并从自噬角度探讨其治疗机制。方法40只SPF级Wistar大鼠随机分为空白组、模型组、阳性对照组和健脾补肾方组,每组10只。除空白组外,其余3组采用腹腔注射氟尿嘧啶和灌胃马利兰溶解液的方法制作AA大鼠模型。造模成功后,健脾补肾方组灌胃给予健脾补肾方,阳性对照组灌胃给予司坦唑醇混悬液,空白组和模型组灌胃给予等体积0.9%氯化钠溶液。连续给药2周。计数2组外周血细胞和骨髓有核细胞数,并检测2组外周血T淋巴细胞亚群、血清细胞因子、骨髓细胞周期、增殖指数(PI)及自噬、凋亡相关蛋白表达。结果阳性对照组和健脾补肾方组外周血Hb、Ret、WBC、PLT、CD3^+、CD4^+、Treg细胞比例、CD4^+/CD8^+、血清IL-11水平、骨髓红系比例、粒系比例、巨核细胞数、骨髓细胞S期和G2/M期细胞比例、PI及LC3-Ⅱ、Bcl-2蛋白表达均明显高于模型组,外周血CD8^+细胞比例、血清IL-2、TNF-α水平、骨髓淋巴细胞比例、骨髓细胞G0/G1期细胞比例及caspase-3、PARP蛋白表达均明显低于模型组,差异均有统计学意义(P<0.05)。结论健脾补肾方能够促进AA大鼠骨髓造血功能恢复,改善外周血象,并增强机体免疫功能,其机制可能与促进骨髓细胞自噬有关。

Objective To investigate the effects of Jianshenbupi prescription on hematopoietic function in rats with aplastic anemia(AA),and to explore its action mechanisms.Methods A total of 40 Wistar rats(SPF)were randomly divided into four groups:blank control group,model group,positive control group and Jianshenbupi prescription group(observation group),with 10 rats in each group.Except for the rats in blank control group,The rats models with AA were established by giving 5-fluorouracil combined with Maryland by gavage.After successful modeling,the rats in observation group were given Jianshenbupi prescription by gavage,and the rats in positive control group were given stanozolol suspension by gavage,and the rats in blank control group and model group were given the same volume of normal saline,for two weeks.And the peripheral blood cells and bone marrow monouclear cells were counted,and the T cell subsets in peripheral blood,serum cytokines,bone marrow cell cycle,PI and the expression levels of autophagy and apoptosis related proteins were detected.Results The levels of Hb,Ret,WBC,PLT,CD3^+,CD4^+,Treg,CD4^+/CD8^+,and the serum levels of IL-11,red cells and megakaryocytes,the proportion of S and G2/M phase cells,PI and the expression levels of LC3-Ⅱand Bcl-2 proteins in positive control group and observation group were significantly higher than those in model group,however,the levels of CD8^+,serum level of IL-2 and TNF-α,the proportion of lymphocytes,the proportion of G0/G1 phase cells and the expression levels of caspase-3 and PARP proteins in positive control group and observation group were significantly lower than those in model group(P<0.05).Conclusion Jianshenbupi prescription can improve the hematopoietic function and immune function of rats with AA,and its action mechanism may be related to the promotion of bone marrow autophagy.