大鼠气虚血瘀肾虚型颈椎病模型的建立_临床医学论文.docVIP

大鼠气虚血瘀肾虚型颈椎病模型的建立_临床医学论文 大鼠气虚血瘀肾虚型颈椎病模型的建立_临床医学论文 作者：王拥军, 施杞 江建春, 梁倩倩 卞琴 李晨光, 周泉, 崔学军, 黄敏, 刘清高, 卢盛, 周重建 【摘要】 目的：采用病、证模型复合的方法建立大鼠气虚血瘀肾虚型颈椎病模型。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血瘀肾虚型颈椎病模型组，每组10只。颈椎病模型采用动静力失衡大鼠颈椎间盘退变模型，气虚血瘀肾虚型颈椎病模型采用动静力失衡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、肾上腺皮质激素和肾上腺素法制造的血瘀模型及去卵巢肾虚模型而制成。通过对大鼠行为体征观察和血液流变学、血小板表面α颗粒膜糖蛋白(alpha，CD62p)、血清雌二醇（estradiol，E2）含量等检测，并用放射免疫法检测血浆环磷酸腺苷（cyclic adenosine monophosphate，cAMP）与环磷酸鸟苷（cyclic guanosine monophosphate，cGMP）含量，验证气虚、血瘀、肾虚证；通过椎间盘组织病理学观察，免疫组织化学法检测椎间盘组织中Ⅱ型、Ⅹ型胶原蛋白含量，实时荧光定量聚合酶链式反应（polymerase chain reaction, PCR)检测颈椎间盘聚集蛋白聚糖（aggrecan，Agc1）、Ⅱ型前胶原基因（type Ⅱ procollagen gene，Col2a1）、基质金属蛋白酶（matrix metalloproteinase，MMP）和基质金属蛋白酶抑制剂（tissue inhibitor of metalloproteinase，TIMP）的基因表达等评判椎间盘退变情况。 结果：病证结合模型大鼠出现精神萎靡、倦怠、舌质瘀紫、尾色瘀青，体质量下降等气虚、血瘀证的表现，cAMP/cGMP降低，低切还原黏度增高，CD62p表达增高，血清E2含量降低。病理提示颈椎间盘组织结构发生退变，Ⅱ型胶原蛋白表达减少，Ⅹ型胶原表达增高，Agc1、Col2a1和TIMP的基因表达降低，MMP表达增高。结论：建立了大鼠气虚血瘀肾虚型颈椎病模型，气虚血瘀肾虚加重了颈椎间盘退变。 【关键词】 气虚; 血瘀; 肾虚; 颈椎病; 病证结合; 动物疾病模型; 大鼠 Objective: To establish a rat model of cervical syndrome with qi deficiency, blood stasis and kidney deficiency. Methods: A total of 30 threeiciency, blood stasis and kidney deficiency group (combined group), with 10 rats in each group. Rats in the normal group received no treatment, rats in cervical syndrome group underwent resection of cervical muscles and ligaments as unbalanced dynamic and static animal model, and rats in combined group underwent resection of both cervical muscles and ovaries, swimming and irregular diet, and injection of adrenal cortex hormone and adrenaline two and a half months after resection as combined model. The qi deficiency, blood stasis and kidney deficiency were determined by observing behaviors and physical signs of the rats, detecting the contents of plasma cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), the hemorrheology, the expression of alphagranular membrane protein (CD62p) and the serum estradiol (E2) content. The aggrecan1, type Ⅱ