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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 serum biochemical markers and trabecular bone structure at different stages in ovariectomized (OVX) rats. Methods Sham group and OVX group of 6-month-old female Sprague-Dawley rats were randomized at 2, 4, 8, and 12 weeks after operation. Four rats were sacrificed at each time point Serum levels of estradiol (E2), bone alkaline phosphatase (BALP) and tartrate-resistant acid phosphatase (TRAP) were measured. Tibial trabecular bone microanalysis was performed using Micro-CT and histological HE staining Structural changes. Results After 2 weeks, E 2 in OVX group began to decline compared with Sham group, and the difference was statistically significant at 4 weeks (P <0.05) until 12 weeks. E 2 level in Sham group did not change with time; After 2 weeks, the peak value reached the peak, then decreased rapidly and dropped to the preoperative level at 12 weeks. The serum TRAP activity in OVX group increased rapidly after operation, reached the peak at 4 weeks after operation and then decreased, still higher than 12 weeks after operation Sham group and preoperative level. Micro-CT analysis was performed on BV / TV, Tb.N, Tb.Sp, Conn.Dn, There was no significant difference in density (BMD) between Sham group at 2 weeks and significant difference at 4 weeks, more significant at 12 weeks and no significant difference in trabecular thickness (Tb.Th). Micro-CT reconstruction At 12 weeks after operation, the trabecular bone of the OVX group was sparse and lacks in severity compared with the Sham group. HE staining showed that the trabecular plate under the epiphyseal epiphyseal plate in the OVX group was thinner and arranged sparsely at 12 weeks after operation, and the trabecular structure was not Mature, poor connectivity between trabecular bone, partially broken, there is a large number of trabecular bone blind, inconsistent trabecular wall thickness. CONCLUSION: The osteoblast and osteoclast activity of OVX osteoporosis model was increased firstly and then decreased, osteoclast relatively longer and longer after osteoporosis, and maintained at a higher level than that of osteoblasts Active, bone resorption than osteogenesis lead to osteoporosis, mainly manifested as trabecular BV / TV, Tb.N, Conn.Dn, BMD decreased, Tb.Sp increased histological manifestations of trabecular structure immature, Trabecular connectivity between the poor, part of the fracture, there are a large number of trabecular bone blind end.