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补肾活血方血清对体外培养兔椎间盘运动节段的影响

[Effect of serum of Bushen Huoxue prescription on rabbit with intervertebral disc motion segments in vitro culture].

作者信息

Zhan Jia-Wen, Zhu Li-Guo, Feng Min-Shan, Wang Shang-Quan, Zhang Ping

机构信息

The Second Department of Spinal, Wangjing Hospital of China Academy of Chinese Medical Sciences, Beijing 100102, China.

The Second Department of Spinal, Wangjing Hospital of China Academy of Chinese Medical Sciences, Beijing 100102, China;

出版信息

Zhongguo Gu Shang. 2018 Jul 25;31(7):627-634. doi: 10.3969/j.issn.1003-0034.2018.07.009.

Abstract

OBJECTIVE

To compare effect of serum of Bushen Huoxue prescription on rabbit with intervertebral disc motion segments in vitro culture.

METHODS

Twenty-four New Zealand white rabbits aged from 4 to 6 months and weighted from 2.5 to 3 kg were divided into medicated group and control group, 7 in each group. Rabbits were excuted under condition of asepsis, 28 spinal motion segments were taken out in each group, and segments were loaded in spinal motion segments in vitro and cultured in culture apparatus. Nutrient solution of medicated group contain 10% serum of Bushen Huoxue prescription, and 10% blank serum in control group. Seven discs between two groups were taken out and observed by histomorphology, proteoglycan PAS/AB stining, collagen II immunohistochemical staining, AGG, Col2aI by PCR test before culture and on the 3th, 7th and 14th day after culture.

RESULTS

Histomorphology results showed the formation of medicated group was better than that of control group at 1 week after culture. PAB/AB test results showed content of poteoglycan between two groups were decreased. Imunohistochemical results showed content of collagen II in medicated group were obviously increased than that of control group at 3 days, but decreased obviously at 1 week than that of control group. PCR results showed expression of Agg in medicated group was obviously decreased than that of control group, but no statistical significance between two groups on the 7 th and 14 th day. Expression of Col2aI between two groups at 3 days were increased, and medicated group increased obviously more, while there were no significant difference between two groups.

CONCLUSIONS

Serum of Bushen Huoxue prescription could delay intervertebral disc degeneration at short time for it is relate with improving circulation, inhibiting inflammatory, regulating extracellular matrix, so the prescription plays an important role in early prevention and treatment for intervertebral disc degeneration.

摘要

目的

比较补肾活血方血清对体外培养的兔椎间盘运动节段的影响。

方法

选取24只4至6月龄、体重2.5至3千克的新西兰白兔,分为用药组和对照组,每组7只。兔子在无菌条件下处死,每组取出28个脊柱运动节段,将节段加载到体外脊柱运动节段培养装置中培养。用药组培养液含10%补肾活血方血清,对照组含10%空白血清。分别于培养前及培养后第3、7、14天从两组中取出7个椎间盘,进行组织形态学观察、蛋白聚糖PAS/AB染色、Ⅱ型胶原免疫组化染色,通过PCR检测AGG、Col2aI。

结果

组织形态学结果显示,培养1周后用药组的结构形成优于对照组。PAB/AB检测结果显示,两组蛋白聚糖含量均降低。免疫组化结果显示,用药组Ⅱ型胶原含量在培养3天时明显高于对照组,但在培养1周时明显低于对照组。PCR结果显示,用药组Agg表达明显低于对照组,但在第7天和第14天两组间无统计学差异。两组在培养3天时Col2aI表达均升高,用药组升高更明显,但两组间无显著差异。

结论

补肾活血方血清可在短期内延缓椎间盘退变,其与改善循环、抑制炎症、调节细胞外基质有关,该方在椎间盘退变的早期防治中起重要作用。

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