L型钙离子通道阻滞剂贝尼地平对去卵巢小鼠骨代谢的研究  

Effects of the l-type calcium channel blocker benidipine on bone formation of ovariectomized mice

作　　者：马忠平[1] 黄健[1] 张志峰[1] 杨云[1] 叶楠[1] MA Zhongping HUANG Jian ZHANG Zhifeng YANG Yun YE Nan(Department of Orthopedics,The Second Affiliated Hospital of Inner Mongolia Medical University, Hohhot 010030, China)

机构地区：[1]内蒙古医科大学第二附属医院关节外科,内蒙古呼和浩特010030

出　　处：《中国骨质疏松杂志》2017年第8期1026-1029,1080,共5页Chinese Journal of Osteoporosis

摘　　要：目的探讨贝尼地平对去卵巢骨质疏松模型小鼠的骨代谢影响。方法随机选取鼠龄10周雌性小鼠30只,均分为假手术组,单纯去卵巢组,去卵巢加药组。假手术组只去除卵巢周围少量脂肪,单纯去卵巢组去除卵巢后灌胃等量生理盐水,去卵巢加药组去除卵巢后灌胃贝尼地平[20 mg/(kg·d),benidipine,BD]3个月。取小鼠股骨,经4%多聚甲醛固定,脱钙后分别进行股骨下段石蜡切片伊红苏木素(HE)染色、转录因子RUNX2(runt-related transcription factor 2,Runx2)免疫组织化学染色及骨钙素(osteocalcin,OCN)免疫荧光(immunofluorescence,IF)染色,并进行显微CT(μCT)扫描。结果股骨下段HE染色发现灌胃贝尼地平的去卵巢小鼠股骨下段骨小梁面积明显高于单纯去卵巢小鼠组(OVX组)(P<0.05),差异具有统计学意义,和假手术组相比,去卵巢加药组股骨下段骨小梁明显降低,差异具有统计学意义(P<0.05);股骨下段RUNX2免疫组化染色发现,和单纯切卵巢组小鼠相比,贝尼地平治疗组的蛋白表达量明显上调,差异具有统计学意义(P<0.05),而各组中,假手术组RUNX2表达量仍然是最高的,差异具有统计学意义(P<0.05);通过骨钙素免疫荧光染色发现,贝尼地平治疗组股骨下段骨钙素表达量明显高于去卵巢组,差异具有统计学意义(P<0.05),但是仍低于假手术组,差异具有统计学意义(P<0.05);此外,通过股骨下段显微CT扫描发现,去卵巢小鼠灌胃贝尼地平组股骨下段骨密度(bone mineral density,BMD),骨小梁厚度(trabecular thickness,Tb.Th),骨小梁数量(trabecular number,Tb.N)均明显高于去卵巢组,差异均具有统计学意义(P<0.05),但骨代谢指标仍低于假手术组,差异均具有统计学意义(P<0.05)。结论贝尼地平能明显减缓由于去卵巢引起的C57/BL6雌鼠骨量丢失,具有比较明显的抗骨质疏松作用。Objective To investigate the effect of Benidipine（ BD） on bone formation in ovariectomized mice. Methods Female mice（ n = 30）,aged 10 weeks,were divided into three groups,include sham,ovariectomy（ OVX） and OVX ＋ BD groups.Sham group received surgical removal of some fat tissue near ovary. OVX group received ovariectomy and intragastric administration of equivalent amount of saline. OVX ＋ BD group received ovariectomy and intragastric administration of BD（ 20 mg/（ kg·d）） for 3 months. At the end of 3 months,mice were killed and distal femurs were taken and fixed by 4% paraformaldehyde and decalcified,then received HE staining,immunohistochemistry（ IHC） staining of RUNX2（ Runt-related transcription factor 2,Runx2）,immunofluorescence staining of osteocalcin（ OCN） and micro-CT（ μCT） scan. Results HE staining showed that distal femur trabecular area increased significantly in the OVX ＋ BD group compared with the OVX group（ P〈0. 05）,but still lower than that of the Sham group（ P〈0. 05）. The immunohistochemistry（ IHC） of RUNX2 at distal femur was also up-regulated in the OVX＋ BD group compared with the OVX group（ P〈0. 05）,the immunofluorescence（ IF） staining of OCN increased in the OVX ＋ BD group compared with the OVX group（ P〈0. 05）,and the maximum expression of RUNX2 and OCN were found in the Sham group（ P〈0. 05）. Moreover,the micro-CT scan of distal femur showed that compared with the OVX group,bone mineral density（ BMD）,trabecular thickness（ Tb. Th） and trabecular number（ Tb. N） all greatly improved in the OVX ＋ BD group（ P〈0. 05）,and the best bone formation index were found in the Sham group（ P〈0. 05）. Conclusion Benidipine promoted bone formation in ovariectomized mice and possessed anti-osteoporosis affect.

关 键 词：骨代谢 骨质疏松 钙通道阻滞剂 绝经期 

分 类 号：R332[医药卫生—人体生理学]