去卵巢建立肾虚型腰椎间盘退变模型的实验研究
[Experimental study on lumbar intervetebral disc degeneration model with kidney deficiency by ovariectomizing].
作者信息
Yao Chang-feng, Zhao Yong-jian, Niu Kai, Sun Yue-li, Li Chen-guang, Tang De-zhi, Shu Bing, Lu Sheng, Zhou Chong-jian, Liang Qian-qian, Shi Qi, Wang Yong-jun
出版信息
Zhongguo Gu Shang. 2013 Dec;26(12):1015-22.
OBJECTIVE
To observe effects of removing arms and ovarian on lumbar intervertebral disc and vertebral bone mineral density (BMD) by establishing rat model of lumbar intervetebral disc degeneration (IDD) with kidney deficiency, and to explore internal mechanism of disc degeneration, relationship between disc degeneration and osteoporosis.
METHODS
Thirty Sprague-Dawley female rats aged one month were randomly divided into control group, lumbar IDD group and lumbar IDD with kidney deficiency group (combined group), 10 rats in each group. Lumbar IDD group removed double arms, lumbar IDD with kidney deficiency group removed double arms after 3 months, both ovaries were removed. Vertebral bone mineral density were observed by Micro-CT scan; morphological changes were tested by safranine O-fast green staining; II, X collagen protein expression in the intervertebral disc were obsevered by immunohistochemistry; extracellular matrix gene expression were obsevered by real-time polymerase chain reaction (RT-PCR), in order to evaluate the effects of removed of forelimbs and double ovarian on degeneration and vertebral bone mineral density of intervertebral disc.
RESULTS
Micro-CT scan showed osteoporosis in kidney deficiency group was obviously worse than other two groups; safranine O-fast green staining showed that intervertebral space became narrowed, intervertebral disc tissue degenerated obviously, chondral palte was underdeveloped in kidney deficiency group; immunohistochemistry showed that X collagen expression increased, type II collagen expression decreased in kidney deficiency group; RT-PCR showed that type II collagen expression in lumbar IDD group and kidney deficiency group was lower than control group, and had statistical meaning among three groups (P=0.000, P=0.000); Age 1 in lumbar IDD group and kidney deficiency group was lower than control group, and had statistical meaning among three groups (P=0.000, P= 0.000); while type X collagen expression was higher than control group, but no significant meaning; MMP-13 in lumbar IDD group and kidney deficiency group was higher than control group, with significant meaning compared among three groups (P= 0.000, P=0.000); aggrecanase-2 in lumbar IDD group and kidney deficiency group was higher than control group, with significant meaning compared among three groups (P=0.006, P=0.008).
CONCLUSION
Rats model of lumbar disc degeneration established by removed forelimbs and ovariectomized can occure "bone like"--osteoporosis, which is similar with clinical kidney lumbar disc degeneration in tissue morphology, molecular cell biology expression.
目的
通过建立肾虚型腰椎间盘退变(IDD)大鼠模型,观察去势及卵巢切除对腰椎间盘及椎体骨密度(BMD)的影响,探讨椎间盘退变的内在机制以及椎间盘退变与骨质疏松的关系。
方法
将30只1月龄雌性Sprague-Dawley大鼠随机分为对照组、腰椎IDD组和肾虚型腰椎IDD组(联合组),每组10只。腰椎IDD组切除双侧上肢,肾虚型腰椎IDD组3个月后切除双侧上肢并切除双侧卵巢。通过Micro-CT扫描观察椎体骨密度;采用番红O-固绿染色检测形态学变化;通过免疫组织化学观察椎间盘Ⅱ型、X型胶原蛋白表达;采用实时聚合酶链反应(RT-PCR)观察细胞外基质基因表达,以评估去前肢及双侧卵巢对椎间盘退变及椎体骨密度的影响。
结果
Micro-CT扫描显示,肾虚组骨质疏松明显重于其他两组;番红O-固绿染色显示,肾虚组椎间隙变窄,椎间盘组织明显退变,软骨板发育不良;免疫组织化学显示,肾虚组X型胶原蛋白表达增加,Ⅱ型胶原蛋白表达减少;RT-PCR显示,腰椎IDD组和肾虚组Ⅱ型胶原蛋白表达低于对照组,三组间差异有统计学意义(P = 0.000,P = 0.000);腰椎IDD组和肾虚组Ⅰ型胶原蛋白表达低于对照组,三组间差异有统计学意义(P = 0.000,P = 0.000);而X型胶原蛋白表达高于对照组,但差异无统计学意义;腰椎IDD组和肾虚组基质金属蛋白酶-13高于对照组,三组间比较差异有统计学意义(P = 0.000,P = 0.000);腰椎IDD组和肾虚组聚集蛋白聚糖酶-2高于对照组,三组间比较差异有统计学意义(P = 0.006,P = 0.008)。
结论
通过切除前肢及卵巢建立的大鼠腰椎间盘退变模型可出现“类骨”——骨质疏松,在组织形态学、分子细胞生物学表达方面与临床肾虚型腰椎间盘退变相似。