Wang Shan Xi, Xie Ju Ying, Xie Xing, Li Shu Zhen
Xiangnan University Medical Imaging Examination and Rehabilitation College, Chenzhou 423000.
Department of Rehabilitation Medicine, Affiliated Hospital of Xiangnan University, Chenzhou 423000.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2021 May;37(3):332-336. doi: 10.12047/j.cjap.6028.2021.002.
To investigate the effects of estrogen receptor α (ERα) gene overexpression on bone metabolism and calcium and phosphorus metabolism in ovariectomized osteoporosis mice, and to provide experimental basis for targeted gene therapy of osteoporosis. Thirty SPF female mice were randomly divided into sham operation group, model group and ERα overexpression group with 10 mice in each group. After the model was established, the ERα overexpression group was transfected with recombinant adenovirus vector carrying mouse ERα gene by intraspinal injection. The model group was transfected with empty virus, and the sham operation group was not treated. The expression of ERα gene in bone tissue of mice was detected by quantitative Real-time PCR (RT-PCR). Bone mineral density (BMD) of mouse femur was measured after modeling. Trabecular number (Tb.N), trabecular thickness (Tb.Th), trabecular segregation (Tb.Sp), bone volume fraction (BV/TV) and biomechanical strength of femur were measured by micro-CT scanning. Serum levels of calcium (Ca), phosphorus (P), osteocalcin (BGP) and alkaline phosphatase (ALP) were measured by automatic biochemical analyzer. The expressions of tissue inhibitor of metalloproteinases 1 (TIMP-1) and monocyte chemotactic protein 1 (MCP-1) in bone homogenate were detected by Immunohistochemistry. Compared with sham operation group, the expression level of ERα gene in bone tissue of model group was decreased significantly, the levels of BMD, BV/TV, Tb. Th, maximum load, rigidity coefficient, Ca and P were decreased, while the levels of Tb. Sp, BGP and ALP were increased significantly (<0.05). Compared with the sham operation group, the expression level of TIMP-1 protein in the bone tissue of the model group was significantly decreased, while that of MCP-1 protein was increased, while that of the ERα overexpression group was increased while that of MCP-1 was decreased (<0.05).The levels of ERα gene expression, BMD, BV/TV, TB. Th, maximum load, rigidity coefficient, Ca and P in the ERα overexpression group were significantly higher than those in the model group, while Tb. Sp, BGP and ALP were significantly lower (<0.05). Compared with the sham operation group, mean optical density of TIMP-1 in the bone tissue of the model group was significantly decreased, while that of MCP-1 was significantly increased, and that of the ERα overexpression group was significantly increased while that of MCP-1 was significantly decreased (<0.05). ERα gene overexpression can improve osteoporosis by regulating bone mineral density, bone parameters, bone metabolism, calcium and phosphorus metabolic indicators and the expression levels of TIMP-1 and MCP-1 in tissues.
探讨雌激素受体α(ERα)基因过表达对去卵巢骨质疏松小鼠骨代谢及钙磷代谢的影响,为骨质疏松的靶向基因治疗提供实验依据。将30只SPF级雌性小鼠随机分为假手术组、模型组和ERα过表达组,每组10只。造模成功后,对ERα过表达组小鼠进行脊髓内注射携带小鼠ERα基因的重组腺病毒载体转染。模型组转染空病毒,假手术组不做处理。采用定量实时荧光定量聚合酶链反应(RT-PCR)检测小鼠骨组织中ERα基因的表达。造模后测量小鼠股骨骨密度(BMD)。通过显微CT扫描测量股骨的骨小梁数量(Tb.N)、骨小梁厚度(Tb.Th)、骨小梁间距(Tb.Sp)、骨体积分数(BV/TV)和生物力学强度。采用自动生化分析仪检测血清钙(Ca)、磷(P)、骨钙素(BGP)和碱性磷酸酶(ALP)水平。采用免疫组织化学法检测骨匀浆中金属蛋白酶组织抑制剂1(TIMP-1)和单核细胞趋化蛋白1(MCP-1)的表达。与假手术组相比,模型组小鼠骨组织中ERα基因表达水平显著降低,BMD、BV/TV、Tb.Th、最大载荷、刚度系数、Ca和P水平均降低,而Tb.Sp、BGP和ALP水平显著升高(P<0.05)。与假手术组相比,模型组骨组织中TIMP-1蛋白表达水平显著降低,MCP-1蛋白表达水平升高,而ERα过表达组TIMP-1蛋白表达水平升高,MCP-1蛋白表达水平降低(P<0.05)。ERα过表达组的ERα基因表达水平、BMD、BV/TV、TB.Th、最大载荷、刚度系数、Ca和P水平均显著高于模型组,而Tb.Sp、BGP和ALP水平显著低于模型组(P<0.05)。与假手术组相比,模型组骨组织中TIMP-1平均光密度显著降低,MCP-1平均光密度显著升高,而ERα过表达组TIMP-1平均光密度显著升高,MCP-1平均光密度显著降低(P<0.05)。ERα基因过表达可通过调节骨密度、骨参数、骨代谢、钙磷代谢指标及组织中TIMP-1和MCP-1的表达水平来改善骨质疏松。