大鼠气虚型颈椎病模型的建立

Establishment of Cervical Spondylosis Model of Qi Deficiency Pattern in Rats

目的:建立大鼠气虚型颈椎病动物模型。方法:选择3月龄雌性SPF级SD大鼠30只,随机分为正常组、颈椎病模型组和气虚型颈椎病模型组,每组10只。颈椎病模型采用动静力失衡性大鼠颈椎间盘退变模型,气虚型颈椎病模型采用动静力失衡性大鼠颈椎间盘退变模型复合疲劳加饥饱失常法制造的气虚模型而制成。通过动物的行为、体征、体质量、环磷酸腺苷(cAMP)、环磷酸鸟苷(cGMP)、乳酸脱氢酶(LDH)来验证气虚证;通过颈椎间盘组织病理学、Ⅱ型和Ⅹ型胶原的免疫组织化学检测及聚集蛋白聚糖(Agc1)、Ⅱ型前胶原基因(Col2 a1)、基质金属蛋白酶-13(MMP-13)和基质金属蛋白酶抑制剂-1(TIMP-1)的基因表达评判椎间盘退变程度,以证实颈椎病模型的成立。结果:与正常组和颈椎病组比较,气虚型组动物出现明显的精神委靡、倦怠、少动、喜卧、毛乱无光泽、眼睑下垂等气虚证的表现,体质量下降;cAMP、cGMP升高,cAMP/cGMP降低(P<0.05,P<0.01),LDH升高(P<0.05);颈椎间盘组织病理学退变更加明显,Ⅱ型胶原蛋白表达减少,Ⅹ型胶原表达增高,Agc1、Col2 a1和TIMP-1基因表达降低,MMP-13表达增高(P<0.05,P<0.01)。结论:通过病-证模型复合的方式可建立大鼠气虚型颈椎病模型,气虚可以加重颈椎间盘的退变。

To establish an animal model of cervical spondylosis （CS） of qi deficiency. Methods ： Thirty three-monthold female Sprague-Dawley rats were randomly divided into normal control group, CS model group and CS of qi deficiency group, with 10 rats in each group. Rats in the normal control group received no treatment, rats of CS model group underwent only resection of cer- vical muscles and ligaments as unbalanced dynamic and static animal model, rats of CS of qi deficiency group underwent both resection of cervical muscles and swimming and irregular diet after two and a half months of the resection. The qi deficiency was determined by observing behaviors and physical signs of the rats, detecting body weight, cAMP, cGMP and LDH, the degeneration of intervertebral discs was determined by detecting the histopathology,the expressions of type Ⅱ collagen and type X collagen proteins, and the expressions of aggrecan-1 （ Agcl ） , type Ⅱ proeollagen gene （ Col2al ） , matrix metalloproteinase-13 （MM-13） and tissue in hibitor of metalloproteinase-1 （ TIMP-1 ） mRNAs. Results ： Compared with normal control group and CS model group, rats of CS of qi deficiency group were noted with obvious signs of tiredness, ptosis, few movement, body weight decrease, cAMP and cGMP increased, cAMP/ cGMP decreased （ P 〈 0.05 or P 〈 0.01 ）, LDH increased （ P 〈 0.05 ）, the degeneration of cervical intervertebral disc morphology were more evident, type Ⅱ collagen protein expression decreased and type X collagen protein expression increased, the expressions of Agcl, Col2al and TIMP-1 mRNA in intervertebral disc also decreased, and the expression of MMP-13 mRNA increased （P 〈0.05 or P 〈0.01 ）. Conclusion： Through combining disease with syndrome, animal model of CS of qi deficiency in rats is established. Qi deficiency can aggravate cervical intervertebral disc degeneration.