大鼠双侧卵巢切除术后不同时期血清骨生化标志物与骨小梁三维结构变化之间的研究  

Relationship between Serum Biochemical Markers of Bone and the Changes of Trabecular Bone Three-dimensional Structure in Different Periods after Rats,Bilateral Ovariectomy

作　　者：马刚[1] 李理[1] 覃永保 李百川[1] 王仁崇[1] 梁伟[1] 吕天成 汤中飞 李兵[1] 

机构地区：[1]广西医科大学第四附属医院/柳州市工人医院,545005

出　　处：《医学研究杂志》2016年第5期76-80,129,共6页Journal of Medical Research

基　　金：国家自然科学基金资助项目(81260273);广西壮族自治区自然科学基金资助项目(2013GXNSFAA019269)

摘　　要：目的观察双侧卵巢切除(OVX)SD大鼠不同时期血清骨生化标志物与骨小梁三维结构变化之间的关系。方法分别于6月龄雌性SD大鼠假手术(Sham)组和双侧卵巢切除术(OVX)组术后2、4、8、12周随机取样,每组每个时间点处死4只,取血清测量雌二醇(E_2)、骨碱性磷酸酶(BALP)、抗酒石酸酸性磷酸酶(TRAP)水平,取胫骨用显微CT(Micro-CT)和组织学HE染色观察胫骨骨小梁微结构变化。结果术后2周,OVX组E_2较Sham组开始下降,4周时差异有统计学意义(P<0.05),一直持续到12周,Sham组E_2水平随时间变化不大;OVX组血清BALP在术后2周达峰值,然后迅速下降,12周时降至术前水平;OVX组血清TRAP活性在术后迅速升高,术后4周达峰值,然后下降,术后12周时仍高于Sham组及术前水平;Micro-CT分析胫骨干骺端相对骨体积(BV/TV)、骨小梁数目(Tb.N)、骨小梁分离度(Tb.Sp)、Conn.Dn、骨密度(BMD)在2周时与Sham组比较差异无统计学意义,第4周开始出现明显差异,12周差异更加明显,骨小梁厚度(Tb.Th)始终无明显差异;Micro-CT重建显示术后12周OVX组骨小梁较Sham组稀疏,缺失严重;HE染色显示术后12周OVX组大鼠胫骨干骺端骨骺板下骨小梁变细且排列稀疏,骨小梁结构不成熟,骨小梁间连接差,部分断裂,出现大量骨小梁盲端,小梁壁厚度不一致。结论 OVX术骨质疏松模型建立过程中大鼠成骨及破骨细胞活跃度先增高后降低,破骨细胞相对而言上升周期更长且下降后能维持在更高水平,比成骨细胞更活跃,骨吸收大于骨生成导致骨质疏松,主要体现为骨小梁BV/TV、Tb.N、Conn.Dn、BMD下降,Tb.Sp升高,组织学表现为骨小梁结构不成熟,骨小梁间连接差,部分断裂,出现大量骨小梁盲端。Objective To observe the relationship between the changes of serum bone biochemical markers and the three - dimensional structure of trabecular bone after rats＇ bilateral ovariectomy. Methods 6 - month - old female SD rats were randomly divided into two groups：sham operated （Sham） group and ovariectomy （OVX） group. At 2 weeks, 4 weeks, 8 weeks,12 weeks after operation, they were randomly sampled, and 4 rats were sacrificed in every group at each time point. Serum levels of estradiol（E2） , bone alkaline phosphatase （BALP） , and tartrate - resistant acid phosphatase（TRAP） were measured. The microstructure changes of trabecular bone in tibia were detected by Micro - CT and hematoxylln - eosin staining（ HE staining）. Results At 2 weeks after operation, E2 in OVX group be- gan to reduce compared with Sham group. It was significantly different from Sham group at postoperative 4 weeks（ P 〈 0.05 ） , which lasted until postoperative 12 weeks. The level of E2 in Sham group was not changed too much with time. Serum BALP activity in OVX group reached a peak at 2 weeks after operation, and then decreased rapidly, equal to preoperative level at postoperative 12 weeks. Serum TRAP activity in OVX group increased rapidly, and reached a peak at 4 weeks after operation, and then decreased, which was still higher than that of Sham group at postoperative 12 weeks and preoperative level. Micro - CT analysis showed no significant difference in relative bone volume （BV/TV） , trabecular number （Tb. N） , trabecular separation （Tb. Sp） , Conn. Dn, bone mineral density （BMD） in OVX group compared with Sham group. At 4 postoperative weeks they began to appear obvious difference, and the difference was more obvious at postoperative 12 weeks. Trabecular thickness （Tb. Th） always had no obvious difference. Micro -CT reconstruction showed that trabecular bone was sparse and seriously missing in OVX group at postoperative 12 weeks compared with Sham group. HE staining showed that the trabecula

关 键 词：卵巢切除术 雌二醇 骨碱性磷酸酶 抗酒石酸酸性磷酸酶 显微CT 苏木精-伊红染色 

分 类 号：R681[医药卫生—骨科学]