引用本文:帅 波,沈 霖,杨艳萍,马 陈,徐晓娟,卢芙蓉,廖 奕.加味青娥丸对模拟失重状态下小鼠骨显微结构和β-catenin及DKK-1表达水平的影响[J].中国临床新医学,2018,11(12):1186-1191.
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加味青娥丸对模拟失重状态下小鼠骨显微结构和β-catenin及DKK-1表达水平的影响
帅 波,沈 霖,杨艳萍,马 陈,徐晓娟,卢芙蓉,廖 奕
430022 武汉,华中科技大学同济医学院附属协和医院中西医结合科
摘要:
[摘要] 目的 观察加味青娥丸对尾吊小鼠发生废用性骨质疏松模型骨显微结构的影响和β-catenin及Diekkopf-1(DKK-1)mRNA表达水平的影响,探讨加味青娥丸防治模拟失重状态下快速骨丢失的部分机制。方法 96只8周龄SPF级C57BL/6小鼠随机分为对照组(control组)、尾吊组(HLS组)、尾吊+加味青娥丸组(HLS+Qing′E组)、尾吊+雌二醇组(HLS+estradiol组),每组24只。control组及HLS组小鼠每天灌服生理盐水0.5 ml,HLS+Qing′E组给予加味青娥丸配制成的0.5 ml混悬液,HLS+estradiol组每天灌服补佳乐溶液0.5 ml,具体实验给药剂量按人与动物体表面积换算法计算。各组小鼠统一条件喂养3个月后,以micro-CT检测并经三维重建获得骨组织微观结构,免疫组化及RT-PCR检测下肢股骨头内骨组织局部β-catenin和DKK-1及其mRNA的表达,ELISA法检测循环血中Ⅰ型原胶原氨基端肽(PINP)的水平。结果 与HLS组比较,control组、HLS+Qing′E组及HLS+estradiol组小鼠骨体积分数(BV/TV)、骨小梁数目(Tb.N)、骨小梁厚度(Tb.Th)显著增高(P<0.05),结构模型指数(SMI)、骨小梁间隙(Tb.Sp)、骨表面积体积比(BS/BV)明显降低(P<0.05)。与HLS组比较,control组、HLS+Qing′E组及HLS+estradiol组小鼠骨组织局部β-catenin mRNA的表达水平明显增高(P<0.05),而DKK-1 mRNA表达水平明显降低(P<0.05)。与HLS组比较,其他三组循环血中PINP水平明显增高(P<0.05)。结论 加味青娥丸能够防治尾吊小鼠发生废用性骨质疏松,其部分疗效机制可能与其增强骨组织局部β-catenin mRNA表达,抑制DKK-1 mRNA水平有关。
关键词:  废用性骨质疏松症  尾部悬吊  模拟失重状态  DKK-1  β-catenin
DOI:10.3969/j.issn.1674-3806.2018.12.03
分类号:R 285.5
基金项目:国家自然科学基金项目(编号:81473492,81403257)
Effects of Modified Qing′ e Pill on bone microstructure and expressions of beta-catenin and DKK-1 in mice under simulated weightlessness
SHUAI Bo, SHEN Lin, YANG Yan-ping, et al.
Department of Integrated Traditional Chinese and Western Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China
Abstract:
[Abstract] Objective To observe the effects of Modified Qing′ e Pill on the bone microstructure and the expression levels of beta-catenin and Diekkopf-1(DKK-1) mRNA in tail suspended osteoporosis model mice, and to investigate its partial mechanisms to prevent and treat rapid bone loss in simulated weightlessness. Methods 96 SPF C57BL/6 mice aged 8 weeks were randomly divided into control group, tail suspension group(HLS group), tail suspension+Modified Qing′ e Pill group(HLS+Qing′e group), tail suspension+estradiol group(HLS+estradiol group), with 24 cases in each group. The control group and HLS group were given 0.5 ml of normal saline every day. The HLS+Qing′e group was given 0.5 ml of mixed suspension of Modified Qing′e Pill, and the HLS+estradiol group was given 0.5 ml of mixed suspension of Bujiale. The doses of the drugs were calculated by the conversion method of human and animal surface area. Three months after treatment, the microstructure of bone tissues was obtained by micro-CT detection and three-dimensional reconstruction. The expressions of local beta-catenin, DKK-1 and its mRNA in the bone tissues of the femoral head of the lower extremities were detected by immunohistochemistry and RT-PCR, and the level of type I procollagen amino end peptide(PINP) in circulating blood was detected by ELISA method. Results Compared with those in the HLS group, the bone volume fraction(BV/TV), bone trabecular number(Tb.N) and trabecular thickness(Tb.Th) increased significantly(P<0.05), and the structural model index(SMI), trabecular spacing(Tb.Sp) and bone surface volume ratio(BS/BV) significantly decreased in the control group, HLS+Qing′e group and HLS+estradiol group(P<0.05). Compared with those in the HLS group, the expression levels of local beta-catenin mRNA increased significantly(P<0.05), and the expression levels of DKK-1 mRNA decreased significantly(P<0.05) in the control group, HLS+Qing′e group and HLS+estradiol group(P<0.05). Compared with those in the HLS group, the levels of PINP in the circulating blood of the other three groups increased significantly(P<0.05). Conclusion Modified Qing′ e Pill can prevent and treat the disused osteoporosis in tail suspended osteoporosis model mice, whose partial mechanisms may be related to the enhancement of the expression of local beta-catenin mRNA and the inhibition of the level of DKK-1 mRNA in bone tissues.
Key words:  Disused osteoporosis  Tail suspension  Simulated weightlessness  Diekkopf-1(DKK-1)  Beta-catenin