中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (15): 2382-2386.doi: 10.12307/2022.595

• 骨科植入物相关基础实验 Basic experiments of orthopedic implant • 上一篇    下一篇

滋肾健骨方干预去势骨质疏松模型大鼠骨微结构及骨密度的变化

林晓生1，韩林静2 ，吴克亮2 ，张  震1，王宏波1，肖庆华1，杜根发2，朱建宗1   

  1. 1深圳市中西医结合医院，广东省深圳市   518104；2广州中医药大学，广东省广州市   510405
  • 收稿日期:2021-05-06 修回日期:2021-05-11 接受日期:2021-06-15 出版日期:2022-05-28 发布日期:2022-01-06
  • 通讯作者: 林晓生，博士，教授，博士生导师，主任医师，深圳市中西医结合医院，广东省深圳市 518104
  • 作者简介:林晓生，2010年广州中医药大学毕业，博士，教授，博士生导师，主任医师，主要从事中西医结合骨质疏松症及其并发症的防治研究。
  • 基金资助:
    广东省中医药局面上科研项目(20191292)，项目负责人：林晓生；深圳市科技创新委员会研究项目(JCYJ20180302144355408，JCYJ20190808100818959)，项目负责人：林晓生；广东省中医药局面上科研项目(20201298)，项目负责人：张震

Effects of Zishen Jiangu Recipe on bone microstructure and bone mineral density in ovariectomized rats with osteoporosis

Lin Xiaosheng1, Han Linjing2, Wu Keliang2, Zhang Zhen1, Wang Hongbo1, Xiao Qinghua1, Du Genfa2, Zhu Jianzong1   

  1. 1Shenzhen Integrative Medicine Hospital, Shenzhen 518104, Guangdong Province, China; 2Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • Received:2021-05-06 Revised:2021-05-11 Accepted:2021-06-15 Online:2022-05-28 Published:2022-01-06
  • Contact: Lin Xiaosheng, Shenzhen Integrative Medicine Hospital, Shenzhen 518104, Guangdong Province, China
  • About author:Lin Xiaosheng, MD, Professor, Doctoral supervisor, Chief physician, Shenzhen Integrative Medicine Hospital, Shenzhen 518104, Guangdong Province, China
  • Supported by:
    Research Projects of Shenzhen Science and Technology Innovation Committee, Nos. JCYJ20180302144355408 and JCYJ20190808100818959 (both to LXS); General Research and Development Projects of Guangdong Provincial Bureau of Traditional Chinese Medicine, Nos. 20191292 (to LXS) and 20201298 (to ZZ)

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摘要：

文题释义：
骨质疏松症：表现为骨微结构退变，骨小梁细小、间距增大，骨密度、骨强度降低，易发骨折的全身性骨病。
滋肾健骨方：为深圳市中西医结合医院院内协定方，该方以“肾虚髓减”为切入点协定，具有补肾益髓、强壮筋骨之效。前期的临床运用中收获良效，但基础研究尚不多，因此在去势大鼠模型中验证滋肾健骨方疗效，对进一步的机制研究意义重大。

背景：滋肾健骨方为深圳市中西医结合医院治疗骨质疏松症的院内协定方，前期在临床中取得了良好的效果，但其治疗骨质疏松症的机制尚不明确。
目的：建立去势骨质疏松大鼠模型，开展体内实验探究滋肾健骨方治疗骨质疏松症的疗效。
方法：80只雌性SD大鼠随机分为手术组68只、假手术组12只，手术组去卵巢建立骨质疏松大鼠模型，术后3个月每组随机抽取6只测骨密度检验造模情况。造模成功后将手术组剩余62只随机分为模型组、中药低、中、高剂量组及阿仑膦酸钠组，中药组分别予以9.375，18.75，37.5 g/(kg·d)的滋肾健骨方，阿仑膦酸钠组予以1.02 g/(kg·d)阿仑膦酸钠，假手术组、模型组予以等体积生理盐水灌胃。干预3个月后麻醉处死各组大鼠，分离胫骨进行Micro-CT骨扫描、苏木精-伊红组织病理染色及骨密度检测，股骨进行三点弯曲试验，ELISA法进行血清碱性磷酸酶、骨钙素及抗酒石酸酸性磷酸酶5b水平测定。此次实验获得广州中医药大学动物伦理委员会批准。
结果与结论：①去势3个月后，手术组大鼠骨密度明显降低，由此证实骨质疏松造模成功；②药物干预3个月后，与模型组相比，中药中、高剂量组、阿仑膦酸钠组血清碱性磷酸酶、骨钙素及抗酒石酸酸性磷酸酶5b水平显著降低，说明滋肾健骨方可降低去势大鼠骨的高转换状态；③干预后中药中、高剂量组骨密度、骨小梁数量、厚度增加，骨小梁间距缩小，骨强度均显著增加，由此证实滋肾健骨方能够改善骨微结构，提高骨密度，降低骨折发生的风险。
缩略语：抗酒石酸酸性磷酸酶5b：tartrate resistant acid phosphatase-5b，TRACP-5b

https://orcid.org/0000-0003-1844-0410 (林晓生) 

中国组织工程研究杂志出版内容重点：人工关节；骨植入物；脊柱；骨折；内固定；数字化骨科；组织工程

关键词: 骨质疏松症, 滋肾健骨方, 骨密度, 骨微结构, 生物力学, 骨代谢

Abstract: BACKGROUND: Zishen Jiangu Recipe is an in-hospital consensus prescription for the treatment of osteoporosis in Shenzhen Integrative Medicine Hospital. It has achieved good clinical results in the early stage, but its mechanism underlying treatment of osteoporosis is still unclear.  
OBJECTIVE: To investigate the therapeutic effect of Zishen Jiangu Recipe on osteoporosis by in vivo experiments in an ovariectomized rat model.
METHODS:  Eighty female Sprague-Dawley rats were randomly divided into a surgery group (n=68) and a sham surgery group (n=12). Rat osteoporosis models were established by ovariectomy inthe surgery group. Three months after the surgery, six rats were randomly selected from each group to test the bone mineral density. After successful modeling, the remaining 62 rats in the surgery group were randomly divided into a model group, a low-dose Zishen Jiangu Recipe group, a medium-dose Zishen Jiangu Recipe group, a high-dose Zishen Jiangu Recipe group, and an alendronate sodium group. Rats in low, medium-, and high-dose Zishen Jiangu Recipe groups were given Zishen Jiangu Recipe at doses of 9.375, 18.75, and 37.5 g/kg per day, respectively. Rats in the alendronate group were given 1.02 g/kg alendronate per day, and rats in the sham surgery and model groups were given an equal volume of normal saline by gavage. After 3 months of intervention, rats in each group were killed after anesthesia, and the tibia was separated for Micro-CT bone scan, hematoxylin-eosin histopathological staining and bone mineral density detection. The femur was taken and subjected to a three-point bending test. And the levels of serum alkaline phosphatase, osteocalcin, and tartrate resistance acid phosphatase 5b were determined by enzyme-linked immunosorbent assay. This experiment was approved by the Animal Ethics Committee of Guangzhou University of Chinese Medicine.  
RESULTS AND CONCLUSION: Three months after ovariectomy, the bone mineral density of the rats in the surgery group was significantly reduced, which confirmed that the osteoporosis model was successfully established. After 3 months of intervention, compared with the model group, the levels of serum alkaline phosphatase, osteocalcin, and tartrate resistance acid phosphatase 5b were significantly reduced in the medium- and high-dose Zishen Jiangu Recipe groups and the alendronate group, indicating that Zishen Jiangu Recipe could reduce the high turnover state of the bone in ovariectomized rats. After the intervention, the bone mineral density, the number and thickness of bone trabeculae were increased, the distance between the bone trabeculae was reduced, and the bone strength was significantly increased in the medium- and high-dose Zishen Jiangu Recipe groups. These findings indicate that Zishen Jiangu Recipe can improve bone microstructure, increase bone mineral density, and reduce the risk of fractures.

Key words: osteoporosis, Zishen Jiangu Recipe, bone mineral density, bone microstructure, biomechanics, bone metabolism

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