二至丸干预去卵巢骨质疏松大鼠骨重建的效应及机制
Effect of Erzhi Wan on Bone Metabolism in Rats with Ovariectomized Osteoporosis
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摘要: 目的 明确二至丸对去卵巢骨质疏松大鼠骨重建的干预作用,并探究其作用机制。方法 采用卵巢切除术制备绝经后骨质疏松症动物模型,确认造模成功后,将60只模型大鼠随机分为模型组、阿仑磷酸钠组、氟化钠组、二至丸组、女贞子组、墨旱莲组等6组,另设假手术组10只。各组分别予:生理盐水(5 mL/kg)、阿仑磷酸钠(1 mg/kg)、氟化钠(5 mg/kg)、二至丸(2 g/kg)、女贞子(1 g/kg)、墨旱莲(1 g/kg)灌胃,1次/日,连续8周,假手术组给药与模型组相同。采用Micro-CT扫描大鼠股骨;双能X线吸收法测定模型股骨颈部位的骨密度(BMD);椎体压缩法检测骨最大载荷量;ELISA法检测血清骨碱性磷酸酶(BALP)、抗酒石酸酸性磷酸酶5b(TRACP-5b)、核因子κB受体活化因子配体(RANKL)、骨保护素(OPG)、β链蛋白(β-catenin)及趋化素(chemerin);qPCR检测骨组织RANKL、OPG、β-catenin及chemerin基因表达。结果 二至丸、女贞子和墨旱莲可显著增加骨量,提升骨最大载荷量,并修复骨微结构。血清中,二至丸、女贞子和墨旱莲可明显降低TRACP-5b、RANKL、chemerin表达水平,并提升BALP、OPG、β-catenin表达水平。骨组织中,二至丸、女贞子和墨旱莲组明显抑制RANKL、chemerin mRNA表达,并促进OPG、β-catenin mRNA表达增加。结论 二至丸可有效改善去卵巢骨质疏松大鼠的骨代谢异常,其作用机制与调控chemerin密切相关。Abstract: OBJECTIVE To clarify the regulating effect of Erzhi Wan (EZW) on bony reconstitution in ovariectomized osteoporosis rats and explore its mechanism. METHODS The animal model of postmenopausal osteoporosis (PMOP) was made by ovariectomy. After confirming the success of the model, 60 model rats were randomly divided into 6 groups, including model group, alendronate sodium group, sodium fluoride group, EZW group, Ligustrum lucidum group and Eclipta group, 10 rats were set up in the sham operation group. Each group was given, physiological saline (5 mL/kg), alendronate sodium (1 mg/kg), sodium fluoride (5 mg/kg), EZW (2 g/kg), Ligustrum lucidum (1 g/kg) and Eclipta (1 g/kg) intragastric administrated once a day for 8 weeks, the intragastric administrated of the sham operation group was the same as that of the model group. Scanning rat femur with Micro-CT; Bone mineral density (BMD) of femoral neck was measured by dual energy X-ray absorptiometry; the maximum load of bone was measured by vertebral compression; Serum bone alkaline phosphatase (BALP), tartrate-resistant acid phosphatase-5b (TRACP-5b), nuclear factor κB receptor activating factor ligand (RANKL), osteoprotectin (OPG), β-catenin and chemerin were detected by ELISA method; qPCR was used to detect bone tissue RANKL, OPG, β-catenin and chemerin gene expression. RESULTS EZW, Ligustrum lucidum and Eclipta significantly increased bone mass, bone maximum load, and repaired bone microstructure.In the serum, EZW, Ligustrum lucidum and Eclipta significantly reduced the expression levels of TRACP-5b, RANKL and chemerin, and increased the expression levels of BALP, OPG, β-catenin. In the bone tissue, EZW, Ligustrum lucidum and Eclipta group significantly inhibited the expression of RANKL and chemerin mRNA and promoted the increase of OPG and β-catenin mRNA expression. CONCLUSION EZW can effectively intervene the abnormal bone metabolism in ovariectomized osteoporosis rats, and its mechanism is closely related to the regulation of chemerin.