不同引经药对股骨头坏死模型兔骨髓干细胞归巢的影响

目的:观察比较活骨Ⅱ方配伍不同引经药对液氮冷冻性股骨头坏死(ONFH)兔股骨头内骨髓干细胞归巢的影响,探讨其防治ONFH的作用机制。方法:84只兔经液氮冷冻法建立股骨头坏死模型后,随机分为模型、活骨Ⅱ方及活骨Ⅱ方分别加牛膝、细辛、独活和桔梗(简称活骨、牛膝、细辛、独活和桔梗)等6组,另14只设为假手术组。在造模同时所有动物均皮下注射rhG-CSF(30μg.kg-1.d-1,连续7 d),并分别灌胃生理盐水和相应中药。于注射rhG-CSF前后分别进行外周血WBC计数。分别于给药2周和4周后,股骨头经HE染色观察组织病理学改变,血管墨汁灌注观察血管形态,免疫组织化学法检测BrdU及SDF-1的表达。结果:与假手术组相比,模型组股骨头空骨陷窝率明显升高,血管面积和SDF-1蛋白表达水平显著降低;与模型组相比,活骨Ⅱ方组股骨头空骨陷窝率降低,血管面积增大、BrdU阳性细胞数和SDF-1表达升高;与活骨Ⅱ方组比较,牛膝组给药4周时股骨头内空骨陷窝率降低、血管面积增大,2周时SDF-1表达增高;桔梗组股骨头内空骨陷窝率有增高趋势,4周时BrdU阳性细胞数明显减少;细辛和独活组无明显变化。结论:活骨Ⅱ方能通过促进动员起来的骨髓干细胞定向归巢从而防治股骨头坏死;引经药牛膝可进一步提高活骨Ⅱ方的上述作用。

Effect of different channel ushering drugs on homing of bone marrow stem cells of femoral head necrosis model rabbits

Objective: To observe the effect of Huogu II formula compatible with different channel ushering drugs on the homing of bone marrow stem cells of osteonecrosis of the femoral head(ONFH) induced by liquid nitrogen freezing in rabbits and discuss the mechanism for preventing and treating ONFH.Method: The ONFH model was established by liquid nitrogen freezing of 84 rabbits.They were randomly assigned to the model group and the Huogu II formula group and groups of Huogu II formula combining with Achyranthis Bidentatae Radix,Asari Radix et Rhizoma,Angelicae Pubescentis Radix,Platycodonis Radix.The remaining 14 rabbits were sham-operated.During the course of ONFH modeling,all of the rabbits were subcutaneously injected with recombinant human granulocyte colony-stimulating factor(rhG-CSF)(30 μg·kg-1·d-1,for consecutively 7 days).Meanwhile,normal saline and decoction of the formulae were orally administrated respectively.WBC was counted in peripheral blood before and after the injection of rhG-CSF.HE stainings at the 2nd and the 4th weeks after the modeling were adopted to observe histopathological changes,vascular morphology was observed by ink perfusion,BrdU and SDF-1 were determined by immunohistochemical assay in femoral heads of the left hind leg.Result: Compared with the sham-operated group,the Huogu II formula group showed decrease in the ratio of empty lacuna and increase in vessel area,number of BrdU positive cells and SDF-1 level.In comparison with the model group,the Achyranthis Bidentatae Radix group displayed decreasing empty lacuna ratio and increasing vessel area at the 4th week and increasing SDF-1 at the 2nd week;the Platycodonis Radix group revealed a notably increasing empty lacuna ratio and a sharp decrease in the number of BrdU positive cells at 4th week;Asari Radix et Rhizoma and Angelicae Dubescentis Radix groups showed no remarkable change.Conclusion: Huogu II formula can promote the directional homing of bone marrow stem cell to the necrosis area.Channel ushering drug achyranthes can further boost above effects of Huogu II formula.