The Study Of Phosphorylation Of Tau Protein In Senile Dementia Based On The Same Disease In Different Syndrome

Objective:To explore the three typical syndromes of the senile dementia, include the syndrome of phlegm obstructing orifice, the syndrome of congealing cold with blood stasis, and the syndrome of insufficiency of kidney essence in six phosphorylation sites difference of Tau proteins phosphorylation(thr205, Thr181, at, ser262, ser404, ser231). Methods:First of all, inherit and improve the modeling methods of this issue, use the theory of TCM etiology combined with modern medicine to construct the senile dementia model, and according the method of the combination of disease and syndrome, superposition related factors in the dynamic process to construct the senile dementia animal model, then on the basis of the disease model, respectively overlay sexual strain and dead tired, feed high fat forage, cold-stress stimulation to build the three typical syndromes of senile dementia, Subsequently, according the test of behavior and blood biochemistry to identify the model was succeed or not.Subsequently, using western blot to test each protein loci’s dephosphorylation. Protein extraction of hippocampus in model rat and protein quantification. Then go through the gums, transfer, rubber, conversion film and antigen-antibody interactions to develop in image analyzer and scan record image. The strip of found by experiment use GelPro 3.2 scan Grayscale and statistics. And then compare the similarities and differences of each target strip.The other part for immunohistochemical experiments to test each protein loci’s dephosphorylation. Using heart perfusion to fix the hippocampus, and then slice pass through the antigen antibody reaction. Under the optical microscope observe the positive changes in each cell synapses. And through the analysis and statistics of IPP software, the results were obtained. Results:1. The rat model of the senile dementia’s disease showed the corresponding performance of blood biochemistry and behavior. Compared with normal group in the test of behavior, the other group escape latent period was prolonged significantly, spent significantly less time in the target quadrant and platform area out traversing-times was decreased, statistically significant(p<0.05). It shows that the study ability of model rats was decreased, conform to the clinical manifestation of senile dementia. While at the blood biochemistry field, the content of testosterone was increased significant in syndrome of kidney deficiency and damage of essence, congealing cold and blood stasis whole blood viscosity was increased significantly at the aspect of hemorheology, the content of blood fat was the highest at the syndrome of phlegm obstructing orifices, all this identified the model was successful.2. In the 262 each syndromes did not show a significant difference. The level of phosphorylation in 181, syndrome of kidney deficiency and damage of essence group was significantly higher than the other group, syndrome of phlegm obstructing orifices group was significantly higher than the other groups; in 205, deficiency of the kidney essence deficiency syndrome group and cold coagulation and blood stasis syndrome group was significantly higher than other groups; in 231, the highest expression of cold coagulation and blood stasis syndrome group, followed by the deficiency of the kidney essence deficiency syndrome group than in other groups also have obvious improvement in 396, cold coagulation and blood stasis syndrome group and other groups of phosphoric acid level and there was no significant difference in 404, syndrome of kidney deficiency and damage of essence group and cold coagulation and blood stasis syndrome group was significantly higher than that of the other groups.3. In the aspect of immunohistochemistry positive area: in 181, syndrome of kidney deficiency and damage of essence group, the positive area was significantly higher than that of other groups, phlegm turbidity obstructing orifices group was significantly higher than the other groups; in 205, syndrome of kidney deficiency and damage of essence group, the positive area was significantly higher than that of other groups; in 231, positive area the highest expression of cold coagulation and blood stasis syndrome group; in 396, cold coagulation and blood stasis group, the positive area expression level was the highest, syndrome of kidney deficiency and damage of essence group was higher than the other group phosphorylation in positive area. In 404, syndrome of kidney deficiency and damage of essence group and cold coagulation and blood stasis group phosphorylation levels were significantly higher than those of the other groups.4. Immunohistochemistry of IOD values, in 181, syndrome of kidney deficiency and damage of essence group phosphorylation of IOD values higher than the value of the other groups; in 205, syndrome of kidney deficiency and damage of essence group positive value was higher than the other groups; in 231, congealing cold blood stasis syndrome group was the highest of IOD value; in 396, cold coagulation and blood stasis group phosphorylation of IOD value expression level was the highest, syndrome of kidney deficiency and damage of essence group was larger than other groups of phosphoric acid of positive area; in 404, cold coagulation and blood stasis group and syndrome of kidney deficiency and damage of essence group phosphorylation of IOD values were higher than those in the other groups. Conclusion:1. It is shown based on experimental results.: syndrome of kidney deficiency and damage of essence group was increased significant, while the phosphorylation of syndrome of congealing cold and blood was mainly focused on the end of C 396 and 404; compared with the pathologic group, phlegm syndrome group mainly in 181 and 231 phosphorylation levels have significantly improved, which indicate that the binding ability of Tau protein and microtubule is reduced in different syndrome models, and it is more important that Tau appears specific phosphorylation.2. The pathological products of the senile dementia is phosphorylation of Tau protein exist in NFT of brain, because of the various pathogenesis, the manifestation of phosphorylation is different. So it is demonstrate the scientific of TCM the same disease with different syndrome theory in the aspect of protein level.3. According to the above results, syndrome of kidney deficiency and damage of essence group of Tau protein phosphorylation occurred at sites are the most widely, and most of the proline rich region, phosphorylation of this site can directly affect the C-terminal phosphorylation, fully shows that syndrome of kidney deficiency and damage of essence may the basic pathogenesis of senile dementia. The maximum amplitude occurred at blood stasis group phosphorylation, and in the C end was the most obvious, it show that the blood stasis may be the ultimate pathology of senile dementia. It is state that the three syndromes are caused the disease each other, and then showed characteristics of TCM are blood stasis due to phlegm, phlegm due to blood stasis, excess syndrome by deficiency and excess syndromes mixed with deficiency.