密骨方对去卵巢骨质疏松模型大鼠骨折愈合过程中神经纤毛蛋白1表达的影响

Effect of Migu Prescription on the expression of neurociliary protein 1 during fracture healing in ovariectomized osteoporosis model rats

目的采用密骨方干预去卵巢骨质疏松骨折大鼠,观察密骨方对骨折断端神经纤毛蛋白1(Nrp-1)表达的影响,探讨中药密骨方影响骨质疏松骨折愈合的机制。方法将3月龄雌性Wistar大鼠随机分为假手术对照组、模型对照组、密骨方治疗组3组适应性饲养1周后采取摘除双侧卵巢的方法建立骨质疏松模型。术后5个月,克氏针右侧股骨骨髓腔固定徒手骨折,并复位固定,建立骨折模型。骨折术后第2天,密骨方组以密骨方灌胃干预。分别在密骨方干预后第2、4、6周采集血液样本及骨组织,用酶联免疫吸附测定法检测大鼠雌激素含量及用Western Blot实验方法检测骨折断端神经纤毛蛋白1含量。结果模型组大鼠骨折断端雌激素含量与假手术组比较显著降低(P<0.05);密骨方组与模型组比较雌激素含量显著升高(P<0.01)。模型组大鼠骨折断端Nrp-1蛋白含量与假手术组比较显著降低(P<0.01);密骨方组大鼠骨折断端Nrp-1蛋白含量与假手术组比较亦显著降低(P<0.01),但与模型组比较,在第4、6周则显著升高(P<0.01)。结论密骨方能提高去卵巢大鼠骨折断端雌激素含量及神经纤毛蛋白1的表达且神经纤毛蛋白1可能作为一个多功能的受体抑制了破骨细胞的生成增强成骨细胞作用,促进绝经后骨折疏松骨折的愈合。

Objective Rats with ovariectomized osteoporosis fracture were adopted to observe the effect of Migu Prescription on the expression of neurociliary protein 1(Nrp1) at the broken end of the fracture,and discuss the mechanism of the effect of Migu Prescription on fracture healing.Methods Three-month-old female Wistar rats were randomly divided into sham operation fracture control group,model control group and Migu Prescription treatment group.After one week of adaptive feeding bilateral ovaries were removed and osteoporosis model was established.Five months after operation,free hand fractures of the right femoral bone marrow cavity fixed Kirschner needle,and the fracture model was established by reduction and fixation.On the second day after fracture surgery,treatment group was given Migu Prescription gavage intervention.Bone tissue samples were collected at the 2 nd,4 th and 6 th weeks after the prognosis of Migu Prescription.Estrogen content in rats was detected by enzyme linked immunosorbent assay and Western Blot was used to detect the content of Nrp1 at the broken end of fracture.Results Compared with sham operation group,the estrogen content in the fracture end of model group was significantly decreased(P<0.05);compared with model group,estrogen content in Migu Prescription group was significantly increased(P<0.05);compared with model group,estrogen content was significantly increased(P<0.01);compared with sham operation group,the Nrp-1 protein content in the fracture end of model group was significantly decreased(P<0.01),and was significantly lower in Migu Prescription group than that in sham group(P<0.01),but compared with model groups,at the 4 th and 6 th week,it was significantly increased(P<0.01).Conclusion Migu Prescription can increase estrogen content and Nrp1 expression at the broken end of fracture in ovariectomized rats,and neurociliated protein 1 may act as a multifunctional receptor to inhibit osteoclast formation and enhance osteoblast action,therefore,promote the healing of postmenopausal fracture loose fracture.