摘要
目的:观察青娥方对去卵巢大鼠骨组织骨桥蛋白(OPN)、骨唾液蛋白(BSP)及其受体整合素α_vβ_3(Integrinα_vβ_3)表达的影响,探讨青娥方防治绝经后骨质疏松症的作用机制。方法:将120只3月龄健康SPF级雌性SD大鼠采用随机数字表法分为假手术组、生理盐水组、青娥方组、骨疏康组,每组30只。除假手术组外,切除大鼠双侧卵巢造模,分别喂服生理盐水(假手术组和生理盐水组)、青娥方和骨疏康,连续给药4、8、12周后取材。ELISA检测血清抗酒石酸酸性磷酸酶(TRACP5b)含量;分离左侧胫骨,制备骨组织切片,HE染色;分离L_(3~5),采用X线骨密度仪检测骨密度;分离L_1、L_2,RT-qPCR、Western Blot分别检测OPN、BSP、Itgα_v、Itgβ_3mRNA及蛋白的表达。结果:卵巢切除组腰椎BMD、骨小梁面积百分比均低于同期假手术组,其中生理盐水组下降幅度最大,术后8、12周有统计学意义(P<0.05或P<0.01)。青娥方组术后12周的腰椎BMD高于同期生理盐水组,差异有统计学意义(P<0.01);青娥方组术后8、12周的骨小梁面积百分比均高于同期生理盐水组,差异有统计学意义(P<0.05)。在3个不同的时间段,卵巢切除组OPN、BSP、Itgα_v、Itgβ_3mRNA和蛋白与血清TRACP5b含量均高于假手术组,其中生理盐水组升高幅度最大(P<0.01);青娥方组5个指标含量均低于同期生理盐水组,除12周的血清TRACP5b含量外,均有统计学意义(P<0.05或P<0.01)。结论:(1)卵巢切除后促进了OPN、BSP及其受体Itgα_v、Itgβ_3的表达,增强破骨细胞性骨吸收,从而导致骨质疏松的发生。(2)青娥方能使骨组织OPN、BSP、Itgα_v、Itgβ_3表达下降,抑制去卵巢大鼠破骨细胞功能活性,从而减少骨质的丢失。
Objective ：To investigate the effect of Qing＇e formula on OPN, BSP and Integrin αvβ3 expression in bone tissue of o- variectomized rats, and further explore the prevention and therapeutic mechanism of Qing＇e formula in osteoporosis of postmenopausal. Methods： 120 of three-months old female SD rats were randomly divided into saline group, Qing＇e formula group, Gushukang group and sham-operated group with 30 rats in each group. All rats except the sham-operated group were ovariectomized and marked as OVX groups. Three days after operation, the rats were fed daily with normal saline （the sham-operated group, the saline group）, Qing＇e formula （the Qing＇e formula group） and Gushukang （the Gushukang group）, respectively. Ten rats were randomly chosen from each group, and sacrificed on week 4, 8 and 12, after treatment. The serum TRACP5b were measured by ELISA. The bone tissues from the left proximal shin were sectioned and processed HE staining. The bone mineral densities （BMD） were measured by dual-energy X-ray absorptiometry （DXA） for the 3-5 lumbar vertebra. The expressions of OPN, BSP, Itgαvβ3 mRNA and protein were measured by quantitative PCR （Q-PCR） and Western Blot, respectively. Results ： The BMD and the area percentage of trabecular of the OVX groups were lower compared with the sham-operated group, of which the saline group were the lowest in week 8 and week 12 （P〈 0.05 or P〈0.01 ）. The BMD of the Qing＇e formula group were higher than that of the saline group in week 12 （P〈0.01）. The area percentage of trabecular of the Qing＇e formula group were higher than that of the saline group in week 8 and week 12 （P〈0.05）. The expression levels of OPN, BSP, Itg, Itgβ3 mRNA and protein along with serum TRACP5b of the OVX groups were higher than the sham-operated group in 4, 8 and 12 weeks, and the saline group showed the highest expression （P〈0.01）. Except the serum TRACP5b in week 12, the expression mentioned above of the Qing＇e formula group was lower than that of the saline group （P〈0.05 or P〈0.01 ）. Conclufion：Ovariectomy promotes the expression of OPN, BSP, Itgα, Itgβ3 which enhance the function of osteoclast, and lead to the occurrence of osteoporosis. The Qing＇e formula can suppress osteoclastic bone resorption by reducing the expression of bone tissue OPN, BSP and its receptor Itgα, Itgβ3. Qing＇e formula plays a positive role in postmenopausal osteoporosis prevention and therapy.
作者
孙雨晴
罗文娟
王柄棋
陈翔
贾晓康
林燕萍
SUN Yuqing LUO Wenjuan WANG Bingqi CHEN Xiang JIA Xiaokang LIN Yanping(Integrative Medicine Institute, Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian 350122, China)
出处
《康复学报》
2017年第3期23-30,共8页
Rehabilitation Medicine
基金
国家自然科学基金项目(81574003)
福建省科技厅引导性项目(2015Y0069)
陈可冀中西医结合发展基金项目(CKJ2014003)
关键词
骨质疏松
去卵巢大鼠
青娥方
整合素
osteoporosis, ovariectomized rat, Qing＇e Fang, integrin
