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目的观察3个老年期痴呆病证结合典型证候模型的大鼠Tau蛋白磷酸化位点的情况,分析总结其磷酸化的特异性。方法间隔1周结扎并剪断左、右颈总动脉,之后腹腔注射D-半乳糖,建立老年期痴呆病大鼠模型,最后在疾病模型的基础上分别进行房劳水劳刺激、投喂高脂饲乳和低温环境刺激,建立3种病证结合(肾虚精亏证、痰浊阻窍证和寒凝血瘀证)的老年期痴呆大鼠模型,通过测定血脂、血流变、血清睾酮水平检查验证各组模型是否成功。随后通过蛋白免疫印迹的方法观察Tau各磷酸化位点在各证候模型中的不同。结果在Tau蛋白脯氨酸富集区,与病理组相比,痰浊阻窍证组主要在205和231位磷酸化水平显著增强,肾虚精亏证组主要在181、205、231位磷酸化水平显著增强,寒凝血瘀证组磷酸化水平增强主要发生在205、231、262位;在C末端区域,与病理组相比,痰浊阻窍证组在404位磷酸化水平显著升高,肾虚精亏证组在396和404位磷酸化水平均显著升高,寒凝血瘀证组两者的升高幅度最高。结论老年期痴呆各典型证候在Tau蛋白磷酸化位点上有特异性,揭示了同病异证的科学内涵。
Objective To observe the phosphorylation sites of Tau in three senile dementia syndromes combined with typical syndrome models and analyze the specificity of phosphorylation. Methods The left and right common carotid arteries were ligated and cut at intervals of 1 week, and then intraperitoneally injected with D-galactose to establish a rat model of senile dementia. Finally, on the basis of the disease model, Animal model of senile dementia with three syndromes combined with syndrome of deficiency of kidney and phlegm, phlegm resistance syndrome and cold coagulation and blood stasis syndrome were established. The levels of serum lipids, blood rheology and serum testosterone were tested and verified Success of each model. Subsequently, the differences of Tau phosphorylation sites in each syndrome model were observed by Western blotting. Results Compared with the pathological group, the levels of phosphorylation at 205 and 231 of the phlegm-dampness-inhibiting syndrome group were significantly increased in the proline-rich region of Tau protein. Phosphorylation at the positions of 180, 205 and 231 The level of phosphorylation in the coagulation-stasis syndrome was mainly at 205,231,262. Compared with the pathology group, the level of phosphorylation at the level of 404 in the phlegm-dampness resistance syndrome group was significantly increased in the C-terminal area, Kidney deficiency essence syndrome group 396 and 404 phosphorylation levels were significantly increased, cold coagulation and blood stasis syndrome both the highest increase. Conclusion The typical syndromes of senile dementia are specific in Tau phosphorylation sites, revealing the scientific connotation of different syndromes.