壮骨止痛胶囊抗绝经后骨质疏松的ERRα作用机制研究

Zhuanggu Zhitong Capsule Protect Against Postmenopausal Osteoporosis Induced by Ovariectomy Via Estrogen Recepor-Related Receptor in Rats

目的通过检测ERRα特异抑制剂XCT790对壮骨止痛胶囊(Zhuanggu Zhitong Capsule,ZGZTC)抗骨质疏松作用的影响,探讨ZGZTC抗骨质疏松作用机理。方法 70只8月龄SD大鼠随机分为假手术组、模型组、XCT790组、ZGZTC组、XCT790高剂量+ZGZTC组、XCT790中剂量+ZGZTC组、XCT790低剂量+ZGZTC组,每组10只,切除卵巢后1周,XCT790组每隔3 d皮下注射XCT790橄榄油溶液,非XCT790组皮下注射无菌橄榄油,ZGZTC组每天灌胃给药1次,连续12周,最后一次给药后1 h腹腔注射水合氯醛麻醉,腹主动脉放血处死,取右股骨检测骨密度和骨生物力学,左股骨ELISA检测ERRα蛋白水平,左胫骨RT-PCR检测ERRαm RNA的表达,右胫骨病理切片免疫组化染色检测ERRα蛋白水平。结果 XCT790显著降低去卵巢大鼠骨密度和骨生物力学强度(P<0.05),轻微降低骨组织ERRα表达水平。ZGZTC显著提高去卵巢大鼠骨密度和骨生物力学强度(P<0.05),且该作用可被XCT790剂量依赖性降低。ZGZTC能显著增强去卵巢大鼠骨组织ERRα表达水平(P<0.05),且这种促进作用可被XCT790明显降低。结论 ZGZTC可通过调节雌激素相关受体信号通路发挥抗骨质疏松作用。

OBJECTIVE To unveil the mechanism of Zhuanggu Zhitong Capsule（ZGZTC） protecting against postmenopausal osteoporosis induced by ovariectomy with XCT790 blocking estrogen receptor-related receptor. METHODS Seventy 8-month old Sprague Dawley female rats were randomly allocated into sham-operation group, model group, XCT790 group, ZGZTC group, XCT790 high dose ＋ ZGZTC group, XCT790 middle dose ＋ ZGZTC group, XCT790 low dose ＋ ZGZTC group, 10 rats in each group. The rats in sham-operation group were retained ovary with only removing some fat near ovary, those rats in other groups were performed bilateral ovariectomy. One week later, the rats in sham-operation group, model group and ZGZTC group were injected subcutaneously with olive oil every 3 days, other rats were injected subcutaneously with XCT790 dissolved in olive oil every 3 days. Rats in sham-operation group, model group, XCT790 group were administered with distilled water, those rats in other groups were administered ZGZTC with the same dosage for 12 weeks. Then all rats were anaesthetized intraperitoneally with injection of chloral hydrate and abdominal aortic exsanguinated. Right femurs were collected for bone mineral density and biomechanics detection, left femurs for ELISA of ERRα, left tibias for RT-PCR of ERRα, right tibias for immunohistochemical detection of ERRα. RESULTS XCT790 significantly decreased bone mineral density and bone biomechanics（P〈0.05） in ovariectomized rats, and decreased the expression of ERRα in bone tissue. ZGZTC could increase the bone mineral density and bone biomechanics（P〈0.05） and the role could be decreased with a dose-dependent pattern by XCT790. ZGZTC significantly enhanced the expression of ERRα（P〈0.05） in ovariectomized rats, and the promoting effect was significantly decreased by XCT790. CONCLUSION ZGZTC plays the role of protecting against postmenopausal osteoporosis through regulating estrogen receptor-related receptor signaling pathway.