摘要
目的：模拟自然界不同强度风寒湿刺激，建立中医风寒湿痹证型颈椎病动物模型。方法：选择24只8月龄雄性新西兰白兔，随机分为正常对照组以及轻、中和重度刺激组。基本刺激条件是6级风力（10．8～13．8m／s），（5±0．5）℃，100％湿度，按轻、中、重度刺激组的不同要求，分别给予32、64和128h的间断重复刺激，每日刺激4h。HE染色，光学显微镜下观察椎间盘组织形态改变，放射免疫法检测前列腺素E2（prostaglandin E2，PGE2）、6-酮-前列腺素F1α（6-ketone—prostaglandin F1α，6-K—PGF1α）和血栓素B2（thromboxane B2，TXB2）含量，免疫组织化学ABC法检测Fas和Bcl-2表达情况，逆转录-聚合酶链反应法检测白细胞介素-1β（interleukin-1β，IL-1β）、肿瘤坏死因子α（tumor necrosis factor α，TNF—α）和转化生长因子β（transforming growth factor β，TGF—β）mRNA表达情况。结果：轻、中度刺激组椎间盘髓核收缩，重度刺激组纤维疏松断裂，软骨终板增厚。与正常对照组比较，轻度刺激组椎间盘组织PGE2有所升高，中度和重度刺激组PGE2、6-K—PGF1α和TXB2升高，IL-1β、TNF—α mRNA和Fas表达明显增加，TGF—βmRNA和Bcl-2表达明显降低。重度刺激组Fas表达进一步增加，Bcl-2表达进一步降低。结论：中、重度风寒湿刺激可以导致家兔颈椎间盘明显退变，可用于建立风寒湿痹证型颈椎病动物模型。
Objective： To make an animal model of cervical spondylosis （arthralgia syndrome type） with stimulation of wind, cold, and dampness.
Methods： Twenty-four 8 months old male New Zealand white rabbits were randomly allocated into four groups： normal control group, light stimulation group, moderate stimulation group and severe stimulation group. The wind speed was 10.8-13. 8 m/s, the temperature was （5±0. 5） ℃, and the humidity was 100%. The rabbits of light, moderate, and severe stimulation groups were kept in the above-mentioned environments for 4 hours everyday, and for a total of 32, 64, and 128 hours, respectively. The intervertebral discs were stained with HE method, and observed with a light microscope. Prostaglandin E2 （PGE2）, 6- ketone-prostaglandin F1α （6-K-PGF1α） and thromboxane B2 （TXB2） contents were measured by ELISA. Fas and Bcl-2 expressions were examined by immunohistochemical avidin-biotin peroxidose complex technique. Interleukin-1β （IL-1β）, tumor necrosis factor α （TNF-α）, and transforming growth factor β （TGF-β） mRNA expressions were examined by reverse transcription-polymerase chain reaction.
Results：The nucleus pulposus of rabbits in the light and moderate stimulation groups shrunken, and in the severe stimulation group, the anulus fibrosus loosed or ruptured, and the cartilage end-plate became proliferated. Compared with rabbits in the normal control group, the PGE2 content rose in the light stimulation group, the contents of PGE2, 6-K-PGF1α, and TXB2 increased, the expressions of IL-1β and TNF-α mRNAs and Fas were up-regulated, and the expressions of TGF-β mRNA and Bcl-2 were down-regulated in the moderate and severe stimulation groups The expression of Fas was up-regulated mostly and Bcl- 2 was down regulated mostly in the severe group.
Conclusion： Moderate and severe stimulations ef wind, cold and dampness can lead to degeneration olcervical intervertebral discs of rabbits. The model corresponds to the theory of traditional Chinese medicine about arthralgia syndrome caused by wind. cold and dampness.
出处
《中西医结合学报》
CAS
2007年第1期39-44,共6页
Journal of Chinese Integrative Medicine
基金
国家自然科学基金资助项目(No.39970917&No.30572398)
上海市基础研究重点资助项目(No.03JC14067)
上海市医学重点学科建设资助项目(No.05Ⅲ027)
关键词
颈椎
风寒湿
痹证
家兔
cervical Vertebrae
Wind cold dampress
arthralgia
syndrome
rabbit
