祛痰逐瘀法对激素性股骨头坏死兔骨髓基质干细胞生物活性的影响

Effects of expelling phlegm and eliminating blood method on biological activity of bone marrow stromal cells in a rabbit model of steroid-induced femoral head necrosis

目的:探讨祛痰逐瘀法对激素性股骨头坏死(ONFH)兔骨髓基质干细胞(BMSCs)生物活性的影响。方法:50只健康普通级新西兰家兔随机分成正常组,模型组,中药低、中、高剂量组,每组10只。连续干预8周,麻醉后取左股骨头行HE染色,无菌下取骨髓培养BMSCs,MTT检测细胞增殖,ALP活性检测成骨能力,茜素红染色观察成骨能力。结果:干预8周后,与正常组比较,模型组空骨陷窝率明显增高,中药高剂量组空骨陷窝率明显低于模型组,中药低、中剂量组(P<0.05)。中药高剂量组BMSCs第72h增殖能力和ALP活性明显高于其它4组(P<0.05)。成骨诱导14d,中药高剂量组茜素红S定量明显高于模型组、中药低剂量组和中药中剂量组(P<0.05),与正常组无显著差异。结论:模型组BMSCs增殖和成骨分化能力下降,中药高剂量组BMSCs增殖与成骨能力明显增强。说明祛痰逐瘀法预防激素性ONFH可能与调控BMSCs成骨分化有关。

Objective： To explore the effect of expelling phlegm and eliminating blood method on biological activity of bone marrow stromal cells（BMSCs） in a rabbit model of steroid-induced osteonecrosis of the femoral head necrosis（ONFH）. Methods： fifty health New Zealand rabbits were randomly divided into 5 groups： control group（N）, model group（M）, Chinese medicine decoction at low dose group（CML）, moderate dose group（CMM） and high dose group（CMH）, ten rabbits per group. After 8-week treatment, all rabbits were anaesthetized to extract the left femoral head for HE staining assay, and collect the bone marrow for culturing BMSCs in accordance to the aseptic manipulation. Subsequently, the cell proliferation was detected by MTT assay and the ability of osteogenesis was detected by ALP activity assay and alizarin red staining. Results： After treatment for 8 weeks, compared with normal group, the percentage of empty lacunae of M group was obviously increased compared to the control group, and this was significantly reversed by treatment with high dose Chinese medicine decoction. The proliferation and ALP activity in BMSCs at the time of 72 hour were significantly enhanced by high dose Chinese medicine decoction compared to the other groups. After osteogenesis-induced for 14 days, the alizarin red staining showed that the ability of osteogenesis was significantly increased by high dose Chinese medicine decoction compared to the M, CML and CMM groups. But, there was no difference in the ability of osteogenesis between CMH and N groups. Conclusion： The ability of proliferation and osteogenesis of BMSCs were significantly decreased osteogenesis-induced, and this effect was improved by high dose Chinese medicine decoction. It indicated that the effect of pretreatment with expelling phlegm and eliminating blood method on ONFH induced by steroid may be related to regulate the differentiation of bone marrow stromal cells.