Bian Qin, Liang Qian-qian, Hou Wei, Zhao Yong-jian, Lu Sheng, Wang Yong-jun, Shi Qi
Institute of Spine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China.
Zhong Xi Yi Jie He Xue Bao. 2010 Feb;8(2):173-80. doi: 10.3736/jcim20100213.
To observe the effects of Qishe Pill, a compound traditional Chinese herbal medicine, on lumbar vertebral bone formation induced by long-time upright posture in rats and to investigate the potential mechanism.
Thirty SD rats were randomly divided into normal control group, untreated group and Qishe Pill group. The rats in normal control group received no treatment and were raised in normal cages. The rats in untreated group and Qishe Pill group were cut off forelimbs by operation so as to be forced to adopt an upright posture for 8 months to induce hyperostosis. Rats in the Qishe Pill group were intragastrically administered with Qishe Pill at a dose of 5 g/(kg x d) for 1 month. All rats were sacrificed at the 9th month after surgery and all lumbar vertebrae were harvested for detection. Safranin O/fast green staining and picrosirius red staining were used to observe pathological changes. Expressions of type I collagen (ColI), type X collagen (ColX), vascular endothelial growth factor (VEGF) and transforming growth factor beta1 (TGF-beta1) in the 5th lumbar vertebra (L(5)) were detected by immunohistochemical method. Expressions of type I collagen alpha2 (Col1alpha2), type X collagen alpha1 (Col10alpha1), TGF-beta1, and VEGF and runt-related transcription factor 2 (Runx2) mRNAs in L(1)-L(3) were detected by real-time fluorescent quantitation reverse transcription-polymerase chain reaction.
Safranin O/fast green staining showed that in the untreated group, non-matrix components increased at marginal lumbar vertebra and intervertebral disc junction, and hyperostosis appeared. However, no obvious change was observed in the normal control group. Non-matrix components decreased at the same location in Qishe Pill group as compared with the untreated group. Picrosirius red staining showed compact collagens at marginal lumbar vertebra and intervertebral disc junction in the normal control group, however, Col I and ColX increased at the same location in the untreated group. In the Qishe Pill group, it showed more compact collagens, especially ColI. Compared with normal control group, expressions of ColX, VEGF and TGF-beta1 were increased at marginal lumbar vertebra and intervertebral disc junction in the untreated group. ColX and VEGF expressions decreased in the Qishe Pill group as compared with the untreated group. Col10alpha1 and Runx2 mRNA expressions were down-regulated by Qishe Pill (P<0.01).
Qishe Pill may delay hyperostosis at marginal lumbar vertebra and intervertebral disc junction, which may be related to reducing type X collagen and Runx2 expressions.
观察复方中药芪麝丸对大鼠长时间直立姿势诱导的腰椎骨形成的影响，并探讨其潜在机制。
将30只SD大鼠随机分为正常对照组、未治疗组和芪麝丸组。正常对照组大鼠不做处理，饲养于正常笼中。未治疗组和芪麝丸组大鼠通过手术切断前肢，使其被迫采取直立姿势8个月以诱导骨质增生。芪麝丸组大鼠按5 g/(kg·d)的剂量灌胃给予芪麝丸，连续1个月。术后第9个月处死所有大鼠，采集全部腰椎进行检测。采用番红O/固绿染色和天狼星红染色观察病理变化。用免疫组化法检测第5腰椎（L5）中Ⅰ型胶原（ColI）、Ⅹ型胶原（ColX）、血管内皮生长因子（VEGF）和转化生长因子β1（TGF-β1）的表达。用实时荧光定量逆转录-聚合酶链反应检测L1-L3中Ⅰ型胶原α2（Col1α2）、Ⅹ型胶原α1（Col10α1）、TGF-β1、VEGF和 runt相关转录因子2（Runx2）mRNA的表达。
番红O/固绿染色显示，未治疗组腰椎边缘和椎间盘交界处非基质成分增多，出现骨质增生；而正常对照组未见明显变化。芪麝丸组上述部位非基质成分较未治疗组减少。天狼星红染色显示，正常对照组腰椎边缘和椎间盘交界处胶原致密，而未治疗组相同部位Col I和ColX增加。芪麝丸组胶原更致密，尤其是ColI。与正常对照组相比，未治疗组腰椎边缘和椎间盘交界处ColX、VEGF和TGF-β1表达增加。芪麝丸组与未治疗组相比，ColX和VEGF表达降低。芪麝丸可下调Col10α1和Runx2 mRNA表达（P<0.01）。
芪麝丸可能延缓腰椎边缘和椎间盘交界处骨质增生，其机制可能与降低Ⅹ型胶原和Runx2表达有关。
