中国组织工程研究

中国组织工程研究 ›› 2021, Vol. 25 ›› Issue (14): 2219-2223.doi: 10.3969/j.issn.2095-4344.3137

• 组织构建与生物活性因子 tissue construction and bioactive factors • 上一篇    下一篇

补肾健脾活血方干预去势大鼠肌肉骨骼转化生长因子β/骨形态发生蛋白2信号通路的变化

刘治军1，刘少津2，魏合伟1，万  雷1，黄宏兴1，乔荣勤1   

  1. 1广州中医药大学第三附属医院，广东省广州市   510405；2广州中医药大学，广东省广州市    510240
  • 收稿日期:2020-04-24 修回日期:2020-04-30 接受日期:2020-06-04 出版日期:2021-05-18 发布日期:2020-12-30
  • 通讯作者: 魏合伟，博士，主任医师，广州中医药大学第三附属医院，广东省广州市 510405
  • 作者简介:刘治军，男，1981年生，广东省阳江市人，汉族，广州中医药大学在读硕士，主治医师，主要从事运动与关节损伤、骨质疏松等方面的研究。
  • 基金资助:
    国家自然科学基金(81674004)，项目负责人：黄宏兴

Effect of Bushen Jianpi Huoxue Recipe on transforming growth factor beta/bone morphogenetic protein 2 signaling pathway in the muscle and skeleton of ovariectomized rats

Liu Zhijun1, Liu Shaojin2, Wei Hewei1, Wan Lei1, Huang Hongxing1, Qiao Rongqin1   

  1. 1The Third Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China; 2Guangzhou University of Chinese Medicine, Guangzhou 510240, Guangdong Province, China
  • Received:2020-04-24 Revised:2020-04-30 Accepted:2020-06-04 Online:2021-05-18 Published:2020-12-30
  • Contact: Wei Hewei, MD, Chief physician, the Third Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • About author:Liu Zhijun, Master candidate, Attending physician, the Third Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • Supported by:
    the National Natural Science Foundation of China, No. 81674004 (to HHX)

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摘要：

文题释义：
补肾健脾活血方：即骨康口服液，是广州中医药大学第三附属医院经验方，主要功效是补益肝肾、强筋健骨。既往研究此方对骨骼的影响较多，然而根据中医脾主肌肉、肾主骨理论，脾为后天之本、肾为先天之本，研究此方对肌肉、骨骼或肌少症与骨质疏松症的关系有较深远的意义。
转化生长因子β：是调控肌肉生长、发育及代谢的重要基因。转化生长因子β信号通路是肌肉骨骼代谢疾病的经典影响通路，对人体免疫、循环、内分泌系统有重大作用，是对多种生物活性都具有调节作用的多肽类超家族，与骨质疏松症的发病密切相关。

背景：补肾健脾活血方是广州中医药大学第三附属医院治疗骨质疏松症的经验方，研究证实转化生长因子β/骨形态发生蛋白2通路能调控骨形成，是参与成骨细胞分化合成的重要通路。
目的：探讨补肾健脾活血方对去势大鼠肌肉骨骼中转化生长因子β/骨形态发生蛋白2信号通路的影响。
方法：6月龄雌性SD大鼠72只，随机分为假手术组24只、模型组48只(去卵巢造模)，术后3个月两组各取12只，检测骨密度。剩余模型组36只随机分为模型组、补肾健脾活血方组(2.979 g/kg)及阿仑膦酸钠组(1.02 mg/kg)，每组12只，2个给药组分别给予相应药物灌胃；假手术组和模型组灌胃等体积生理盐水。12周后麻醉处死各组大鼠，双能 X 射线法检测大鼠腰椎和股骨骨密度，实时荧光定量聚合酶链式反应检测大鼠肌肉骨骼中转化因子β、骨形态发生蛋白2 mRNA的表达，蛋白免疫印迹法检测转化生长因子β、骨形态发生蛋白2蛋白的表达。实验方案获得广州中医药大学伦理委员会认可(批准编号：S2016046)。
结果与结论：①造模3个月后，与假手术组比较，模型组大鼠骨密度明显降低(P < 0.05)；药物干预3个月后，与模型组比较，补肾健脾活血方组及阿仑膦酸钠组骨密度均提高明显，且转化生长因子β、骨形态发生蛋白2 mRNA和蛋白表达水平显著提高(P < 0.05)；②结果说明，补肾健脾活血方通过提高转化生长因子β、骨形态发生蛋白2 mRNA和蛋白表达水平，来调控转化生长因子β/骨形态发生蛋白2信号通路的转导，进而对绝经后骨质疏松症起防治作用。

https://orcid.org/0000-0002-6624-0408 (刘治军) 

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 骨质疏松, 骨密度, 中药, 补肾健脾活血方, 转化生长因子β, 骨形态发生蛋白2, 信号通路, 大鼠

Abstract: BACKGROUND: Bushen Jianpi Huoxue Recipe is an empirical prescription for treating osteoporosis in the Third Affiliated Hospital of Guangzhou University of Chinese Medicine. Studies have confirmed that the transforming growth factor β (TGF-β)/bone morphogenetic protein 2 (BMP-2) signaling pathway can regulate bone formation, which is an important pathway for the differentiation and synthesis of osteoblasts. 
OBJECTIVE: To investigate the effect of Bushen Jianpi Huoxue Recipe on TGF-β/BMP-2 signaling pathway in the muscle and skeleton of ovariectomized rats. 
METHODS: Seventy-two SPF female Sprague-Dawley rats aged 6 months were randomly divided into sham operation group (n=24) and model group (n=48). In the model group, ovariectomized models were made in rats. Twelve rats from each group were randomly selected 3 months after operation for testing bone density. The remaining 36 rats in the model group were randomly divided into model group, Bushen Jianpi Huoxue Recipe group (BSJPHX group, dose 2.979 g/kg), and alendronate sodium D3 tablets (II) group (ALN group, dose 1.02 mg/kg). The sham operation group and the model group were given an equal volume of normal saline. After 12 weeks, rats in each group were sacrificed under anesthesia. The bone density of lumbar vertebrae and femur were detected by dual-energy X-ray method. Real-time quantitative polymerase chain reaction (real-time PCR) and western blot were used to detect TGF-β and BMP-2 mRNA and protein in rat muscle and bone, respectively. The experiment was approved by the Ethics Committee of Guangzhou University of Chinese Medicine, with an approval No. S2016046. 
RESULTS AND CONCLUSION: After 3 months of modeling, the bone mineral density of the model group was significantly lower than that of the sham operation group (P < 0.05). After 3 months of drug intervention, compared with the model group, bone mineral density was significantly increased in the BSJPHX group and ALN group, and TGF-β and BMP-2 mRNA and protein expression levels were significantly increased (P < 0.05). To conclude, Bushen Jianpi Huoxue Recipe regulates the transduction of TGF-β/BMP-2 signaling pathway by increasing the expression of TGF-β and BMP-2 at mRNA and protein levels, and then plays a preventive role in postmenopausal osteoporosis.

Key words: osteoporosis, bone mineral density, traditional Chinese medicine, Bushen Jianpi Huoxue Recipe, transforming growth factor β, bone morphogenetic protein 2, signaling pathway, rat

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