青娥方对去卵巢大鼠血清MMP-9、TRACP5b、Cath-K及骨组织形态结构的影响

Effect of Qing'e Fang on ovariectomized rats serum MMP-9、TRACP5b、Cath-K and bone tissue morphology

目的观察青娥方对卵巢切除大鼠血清基质金属蛋白酶-9(MMP-9)、抗酒石酸酸性磷酸酶5b(TRACP5b)、组织蛋白酶K(Cath-K)及骨组织形态结构的影响,进一步揭示青娥方防治绝经后骨质疏松症的作用机制。方法将SD雌性大鼠随机分为假手术组、生理盐水组、青娥方组和骨疏康组。除假手术组外,运用手术切除大鼠双侧卵巢造模,并分别灌服生理盐水(假手术组和生理盐水组)、青娥方和骨疏康,干预4、8、12周后分批处死取材,ELISA法检测血清MMP-9、TRACP5b、Cath-K及尿DPD等含量,除蛋白法检测尿Cr含量,HE染色法观察胫骨上段形态并计量。结果与同期假手术组比较,生理盐水组大鼠血清MMP-9、TRACP5b、Cath-K含量及DPD/Cr比值均明显提高(P<0.01,P<0.05);与同期生理盐水组比较,青娥方组和骨疏康组大鼠血清MMP-9、TRACP5b、Cath-K含量及DPD/Cr比值均下降,除4周的血清MMP-9含量外,均具统计学意义(P<0.01,P<0.05)。青娥方组骨小梁数目、排列以及结构完整性均优于同期生理盐水组,Tb.Ar%在8、12周较同期生理盐水组明显提高(P<0.01,P<0.05)。结论青娥方能通过减少血清中MMP-9、TRACP5b、Cath-K的含量,抑制破骨细胞的功能活性,削弱对骨基质的吸收破坏,发挥护骨强骨之功。

Objective： To observe the effect of Qing’e Fang serum matrix metalloproteinase-9（MMP-9）, tartrate resistant acid phosphatase 5 b（TRACP5 b）, cathepsin K（Cath-K） and bone tissue morphology on ovariectomized rats, further reveal the Qing’e Fang in treating postmenopausal osteoporosis mechanism. Methods： SD rats were randomly divided into sham operation group, saline group, Qing’e Fang group and Gushukang group. Except the sham operation group, the use of surgical resection of bilateral ovaries of rats model, and treated with saline（sham operation group and saline group）, Qing’e Fang and Gushukang, respectively and sacrificed with intervention 4 weeks, 8 weeks, 12 weeks were sacrificed, ELISA was applied to detect serum MMP-9, TRACP5 b, Cath-K and DPD in urine the content, in addition to the detection of urinary Cr protein content, observation of tibial morphology and measuring HE staining method. Results： Compared with the sham operation group, saline group rats serum MMP-9, TRACP5 b, Cath-K content and DPD/Cr ratio were significantly increased（P〈0.01, P〈0.05）, compared with the normal saline group, Qing’e Fang group and Gushukang group rats serum MMP-9, TRACP5 b, Cath-K content and DPD/ratio was Cr in addition to decrease the content of serum MMP-9 of 4 weeks, were statistically significant（P〈0.01, P〈0.05）. The number of Qing’e Fang group, bone trabeculae and structure integrity are better than the normal saline group, Tb.Ar% increased significantly at 8 and 12 weeks compared with saline group（P〈0.01, P〈0.05）. Conclusion： Qing’e Fang can reduce the content of serum MMP-9, TRACP5 b and Cath-K, inhibit osteoclastic function, weakening of bone matrix resorption and bone support play strong bone.