Zhu Li-Guo, Zhang Ping, Song Qing-Hui, Zhang Wan-Qiang, Zhu Hong-Wei, Yang Li-Ping, Zhu Yan, Wang Yuan, Li Ling-Hui
Department of Pharmacology, Wangjing Hospital of China Academy of Traditional Chinese Medical Scinences, Beijing 100102, China;
Zhongguo Gu Shang. 2017 Oct 25;30(10):926-932. doi: 10.3969/j.issn.1003-0034.2017.10.010.
To study the effect of Bushen Huoxue decoction on calcification of cartilage endplate in lumbar vertebrae.
Six healthy male gerbils with 2-month-old were selected as normal control group, and 24 7-month-old healthy male gerbils were fed to 12-month-old to establish the aged gerbil model. Thirty gerbils were randomly divided into five groups as follow: the normal control group (=6), model group (=6, normal saline 4 ml/kg, intragastric 30 d), Bushen Huoxue low dose group(=6, 1.9× 10⁻ ³ ml/g given Bushen Huoxue recipe orally, 30 d), Bushen Huoxue middle dose group(=6, 3.8× 10⁻ ³ ml/g given Bushen Huoxue recipe orally, 30 d), Bushen Huoxue high dose group(=6, 7.6× 10⁻ ³ ml/g given Bushen Huoxue recipe orally, 30 d), the intervention group administered for 1.36 g from 7-month-old age, 30 d. The animals were sacrificed at the age of 2 months in the normal control group and 12 months of age in the other groups. The morphology of the lumbar vertebral cartilage endplate, the area of vascular bud, the ratio of non-calcified/calcified layer were analysis by HE chromosome visual method. The expression of type X collagen and BMPs in cartilage endplates were detected by rabbit monoclonal immunohistochemical staining.
The relative area of the vascular buds cartilage endplate measurements showed that compared with the model group, middle dose group and normal control group increased (<0.05), high and low dose groups all had different degrees of increase, but no statistical significance(>0.05). The ratio of cartilage endplate thickness of non-calcified/calcified showed that compared with the model group, Bushen Huoxue middle dose, normal control group increased, with statistical significance(<0.05), and high and low dose groups all had different degrees of increase, but there were no statistical significance(>0.05). Compared with the model group, the expression of type X collagen in the cartilage endplate of the normal group, the Bushen Huoxue low, middle and high dose groups decreased, and had statistical significance(<0.01); compared with the model group, the expression of BMPs in the normal group, Bushen Huoxue middle dose group increased, with statistically significant(<0.01), while the high and low dose groups increased in different degrees, but there was no statistical significance(>0.05).
Bushen Huoxue prescription can delay the calcification of cartilage endplate in the process of aging, suggesting that it can be used as a preventive medicine for early disc degeneration.
研究补肾活血汤对腰椎软骨终板钙化的影响。
选取6只2月龄健康雄性沙鼠作为正常对照组,将24只7月龄健康雄性沙鼠饲养至12月龄建立老年沙鼠模型。30只沙鼠随机分为5组:正常对照组(n = 6)、模型组(n = 6,生理盐水4 ml/kg,灌胃30 d)、补肾活血低剂量组(n = 6,给予补肾活血方1.9×10⁻³ ml/g灌胃,30 d)、补肾活血中剂量组(n = 6,给予补肾活血方3.8×10⁻³ ml/g灌胃,30 d)、补肾活血高剂量组(n = 6,给予补肾活血方7.6×10⁻³ ml/g灌胃,30 d),干预组从7月龄开始给药1.36 g,30 d。正常对照组2月龄处死动物,其他组12月龄处死动物。采用HE染色观察法分析腰椎软骨终板形态、血管芽面积、非钙化/钙化层比值。采用兔单克隆免疫组化染色检测软骨终板中X型胶原蛋白和骨形态发生蛋白(BMPs)的表达。
软骨终板血管芽相对面积测量结果显示,与模型组相比,中剂量组和正常对照组增加(P<0.05),高、低剂量组均有不同程度增加,但无统计学意义(P>0.05)。软骨终板非钙化/钙化厚度比值显示,与模型组相比,补肾活血中剂量组、正常对照组增加,有统计学意义(P<0.05),高、低剂量组均有不同程度增加,但无统计学意义(P>0.05)。与模型组相比,正常组、补肾活血低、中、高剂量组软骨终板中X型胶原蛋白表达降低,有统计学意义(P<0.01);与模型组相比,正常组、补肾活血中剂量组BMPs表达增加,有统计学意义(P<0.01),高、低剂量组不同程度增加,但无统计学意义(P>0.05)。
补肾活血方能延缓衰老过程中软骨终板的钙化,提示其可作为椎间盘早期退变的预防用药。