化瘀补肾方对卵巢切除小鼠骨折愈合的作用

目的观察化瘀补肾方对卵巢切除小鼠骨折愈合的作用。方法 3月龄雌性C57BL/6小鼠48只分为假手术组、模型组和化瘀补肾方组各16只。模型组和化瘀补肾方组小鼠采用双侧卵巢切除术(OVX)建立骨质疏松症模型。术后12周,各组小鼠建立左侧胫骨中段骨折模型。术后给予化瘀补肾方组小鼠化瘀补肾方灌胃,给予假手术组与模型组小鼠等体积生理盐水灌胃,至骨折术后4周取材。检测各组小鼠血清雌二醇水平,采用Micro-CT检测小鼠第4腰椎及左侧胫骨的骨密度,检测左侧胫骨最大应力,对左侧胫骨石蜡切片进行阿尔辛蓝/橙黄G染色,观察骨组织形态学变化。结果 OVX后4周开始,模型组小鼠体重明显升高,并持续高于假手术组(P0.01)。骨折术后4周,模型组小鼠血清雌二醇和第四腰椎骨密度(BMD)明显低于假手术组(P0.01),化瘀补肾方组小鼠血清雌二醇水平和第四腰椎BMD高于模型组(P0.01)。Micro-CT三维重建显示,模型组左侧胫骨仍有明显的骨折线,骨痂生长不明显,化瘀补肾方组骨折线模糊,骨痂形成明显。与假手术组相比,模型组小鼠的BV/TV(P0.01)、Tb.Th(P0.05)和Tb.N(P0.01)均明显降低,Tb.Sp明显升高,化瘀补肾方组小鼠的BV/TV(P0.05)、Tb.Th(P0.05)相较于模型组则显著升高,但Tb.N、Tb.Sp差异无统计学意义。与假手术组比较,模型组小鼠胫骨的最大应力在骨折术后28 d时显著低于假手术组(P0.01),化瘀补肾方组小鼠胫骨的最大应力显著高于模型组(P0.01)。阿尔辛蓝/橙黄G染色显示,假手术组和化瘀补肾方组骨痂组织处于塑形期,可见明显骨小梁结构,而模型组骨痂组织内见大量脂肪组织,骨小梁稀疏。结论化瘀补肾方能有效促进去卵巢小鼠的骨折愈合,改善愈合骨组织的生物力学性能。

Effects of Huayu Bushen Formula on bone fracture healing in ovariectomized mice

Objective To observe the effect of Huayu Bushen Formula on fracture healing in ovariectomized mice. Methods 36 female C57BL / 6 mice of 3 month old were randomly divided into sham group, model group and Huayu Bushen Formula group with 12 mice in each group. In the model group and Huayu Bushen formula group, the osteoporosis model was established by ovariectomy (OVX). Twelve weeks after OVX, the left middle tibial fracture model was established in all the mice. After operation, mice in Huayu Bushen Formula group were given Huayu Bushen formula by intragastric administration, and mice in sham group and model group were given equal volume of normal saline. The serum estradiol content was measured 4 weeks after fracture operation. The bone mineral density (BMD) of the fourth lumbar (L4) vertebrae and the left tibias were measured by Micro-CT, and the moment of inertia of the left tibia was calculated. The paraffin sections of the left tibias were stained with Alcian blue/Orange G to observe the morphological changes of the callus. Results From 4 weeks after OVX, the body weight of the model group increased significantly, and continued to be higher than that of the sham operation group (P<0.01). The serum estradiol level and L4 BMD of model group were significantly lower than that of sham group (P<0.01), and were increased in Huayu Bushen Recipe group compared with those of model group (P<0.01). The 3D reconstruction images of left tibias showed that obvious fracture line in the model group but not in sham group and Huayu Bushen Formula group. Micro-CT analysis also showed that compared with sham group, the BV/TV(P<0.01), Tb.Th (P<0.05) and Tb.N (P<0.01) of callus of model group were significantly lower, and Tb.Sp was increased (P<0.01); while BMD (P<0.01), BV/TV (P<0.05) and Tb.Th (P<0.05) of the Huayu Bushen Formula group were significantly higher than those of the model group. There was no significant difference in Tb.N and Tb.Sp between the two groups. Compared with the sham group, the moment of inertia of callus in the model group was significantly higher (P<0.01), while it was significantly decreased in the Huayu Bushen Formula group compared with that of the sham operation group (P<0.05). Alcian blue/Orange G staining showed that in the sham group and Huayu Bushen Formula group, the callus tissue was in the shaping stage with trabecular bones in it, while there was plenty of adipose tissue in the callus of the model group with very few trabecular bone formation. Conclusion Huayu Bushen Formula can effectively promote fracture healing in ovariectomized mice, and improve the biomechanical properties of bone callus.