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【目的】探讨组织蛋白酶K(cathepsin K,CK)在骨质疏松症发病中的意义,观察补肾通络方对去卵巢大鼠破骨细胞CK表达的影响。【方法】选用30只雌性SD大鼠随机分成3组:假手术组、模型组及补肾通络方组(中药组,剂量为12.46 g.kg-1.d-1);后2组切除双侧卵巢,4周后中药组给予补肾通络方灌胃。分别于第6、12周测定各组大鼠全身骨密度(BMD),并采用Western blot及逆转录—聚合酶链反应(RT-PCR)法测定体外分离培养破骨细胞中CK的蛋白及基因表达。【结果】体外分离培养椎骨来源的破骨细胞具有成熟破骨细胞的表型特征,去卵巢模型大鼠骨密度在时效上与CK的表达呈现负相关;补肾通络方以时效方式下调CK蛋白及CK mRNA的表达水平(均P<0.05)。【结论】CK的过度表达是导致骨密度下降的重要原因,是骨质疏松症发生的潜在危险因子;以时效方式下调CK蛋白及CK mRNA的表达进而治疗骨质疏松症可能是补肾通络方的作用机制之一。
【Objective】 To investigate the significance of cathepsin K (CK) in the pathogenesis of osteoporosis and to observe the effect of Bushen Tongluo Recipe (CKD) on CK expression in osteoclasts of ovariectomized rats. 【Methods】 Thirty female Sprague-Dawley rats were randomly divided into three groups: sham-operation group, model group and Bushengtongluo Recipe group (Chinese medicine group, dose of 12.46 g.kg-1.d-1) Ovarian side, 4 weeks after the Chinese medicine group given Bushen Tongluo gavage. Whole body bone mineral density (BMD) was measured at the 6th and 12th week in each group, and the protein and gene of CK in osteoclasts cultured in vitro were determined by Western blot and reverse transcription polymerase chain reaction (RT-PCR) expression. 【Results】 The osteoclasts derived from vertebra were isolated and cultured in vitro with the phenotypic characteristics of mature osteoclasts. The bone density of ovariectomized rats showed a negative correlation with CK expression in time-dependent manner. Bushen Tongluo Decoction could down-regulate CK protein And CK mRNA (all P <0.05). 【Conclusion】 Overexpression of CK is an important factor leading to the decrease of bone mineral density and a potential risk factor for osteoporosis. To treat osteoporosis by down-regulating the expression of CK protein and CK mRNA may be Bushen Tongluo Recipe One of the mechanisms of action.