Effects Of Abnormal Cholesterol Metabolism On Aβ In Patients With Coronary Atherosclerosis And The Study Of Compound Intervention

Purpose: Establishment a pig animal model of highcholesterol diet induced A beta increased Bama miniature through the high cholesterol, to observe the effect of hypercholesterolemia state of beta amyloid production within the brain, to explore the relationship between the related factors of cerebral pathological disorders of cholesterol metabolism in hypercholesterolemic condition with Alzheimer’s disease early beta amyloid generation; with the different efficacy of traditional Chinese medicine intervention in the animal model, and to explore its mechanism.By reducing the typical pathological features of ad A-beta generation and deposition to prevention and treatment of AD in early stage and to provide the experimental basis.Material and method:Guangxi Bama miniature pig 55, 6-8 months, weight(29.88 + 2.27 kg, male, intestinal bacteria group normal, healthy, by Taizhou Taihe Biological Technology Co., Ltd.. Ethics in line with the Ministry of health of the people’s Republic of China Animal Experimental Management Regulations(No.552001) and the experimental animal regulations of Liaoning University of Traditional Chinese Medicine regulations. The laboratory temperature was(20.80 + 2.97) and humidity(47.30 + 15.31)% and the experiment animals were single cage feeding.. After 1 weeks of feeding, 55 small pigs were divided into two groups by random digits table method: normal group 5, operation group 50. Normal group was fed with basic diet, operation group(2nd week of coronary balloon compression with strain, preoperative fasting for 8 h water deprivation for 2 hours), postoperative continue to be high in calories and fat amount bile salt feeding(to prevent pig continued to eat lots of greasy food in anorexia, and bile salts is to prevent indigestion). The daily feed intake was 3% of the weight of the last week, free drinking water. There were 8 small pigs died in operation;According to the random number table survived 42 pigs were re randomized: model group(n = 7), invigorating the spleen and removing phlegm and blood stasis group(n = 7), Jianpi Qutan group(n = 7), invigorating the spleen group(n = 7), eliminating phlegm and removing blood stasis group(n = 7), western medicine group(n = 7) so far, in the group of invigorating the spleen to join Jianpi Recipe, Jianpi Qutan joined the group of Jianpi Qutan recipe, and removing phlegm and blood stasis group to join the eliminating phlegm and removing blood stasis, invigorating the spleen and removing phlegm and blood stasis compound group join Jianpi Qutan Huayu Decoction, add appropriate dose of traditional Chinese medicine according to the weight of the corresponding standard, were fed to 24 weeks, the indexes are as follows:1 During the experiment, the weight of the mini pig was weighed every 4 weeks, and the weight of the pig was observed.. 2. The 0 week and experiments began after 4 weeks, 12 weeks, 24 weeks fasting 16 h after auricular vein blood 3ml, full automatic biochemical analyzer was used to measure the blood lipid(TC, TG, LDL, HDL) levels observed four changes in the blood at different time points. 3.the distribution of A beta cells in brain tissue was detected by immunohistochemistry staining.. 4. Immunohistochemical method to observe the content and level of A beta in brain tissue. 5 The expression of A beta metabolism related factor BACE1 and brain cholesterol metabolism factor HMG-Co AR, CYP46 a, ABCA1 in brain were detected by immunohistochemistry.. 6 The expression of A beta metabolism related factor BACE1、 and brain cholesterol metabolism related factor Apo E, CYP46 a, ABCA1 and HMG-Co AR were detected by Blot Western. The results of this experiment were statistically analyzed by using SPSS17.0 software, and the results were compared with the standard deviation(X + S).The single factor analysis of variance was used in the comparison between the groups and the group..(P<0.05 shows that there are significant differences, and P<0.01 shows a very significant difference.)Results:1.. Normal group and experimental mini pig model of gaining weight change display: model group and normal group in the 0 week, the body weight growth, there was no significant difference, two groups of Bama miniature pig weight growth from 4 weeks there was significant difference(P < 0.05); 2. Changes of blood lipid was not obvious of normal piglet in the growth process,and model group Bama miniature pig blood lipid change significantly. The results showed that: high fat diet for 1 week, the miniature pig elevated triglycerides 2.86 times, 3.84 times increase in the total cholesterol, low density lipoprotein 7.40 times higher, the high density lipoprotein increased 2.56 times; with the extension of a high-fat diet, miniature pig triglyceride(TG), total cholesterol(TC), low density lipoprotein(LDL) continues to rise, the 24 week tends to be stable, and high density lipoprotein with a high-fat diet increased decreased; 3. Immunohistochemistry method showed high blood cholesterol levels increased brain A levels, Bama small pig A model group were significantly higher than those in normal control group beta(0.26 + 0.03 + 0.01 pg/mg pg/mg protein vs 0.1110 protein, p<0.01). Immunohistochemistry method was used to detect the average optical density(AOD) of normal control group compared with the model group, there was statistical significance.(P < 0.01). The immunohistochemical staining showed that compared with normal control group, model group of brain neurons can be seen clearly brown granules, said the increased expression of A positive cells in the brain tissue of beta; 4.Group of Strengthening the spleen and removing phlegm and Blood Stasis Decoction to adjust the TC, HDL, LDL is better than the other two groups, group of invigorating spleen and removing phlegm and blood stasis regulating TG abilityis better than the other two groups. 5. Compared with the model group, group of invigorating the spleen, group of dispelling phlegm, group of eliminating phlegm and removing blood stasis, group of invigorating the spleen and removing phlegm and blood stasis can significantly reduce brain Abeta content(P < 0.01); Jianpi Qutan Huayu group than in the group of invigorating the spleen and dispelling phlegm group, and removing phlegm and blood stasis group reduced content of amyloid beta, and there is statistical significance(P < 0.01), there was no significant difference between the group of invigorating the spleen and dispelling phlegm and group of emoving phlegm and blood stasis(P > 0.05); 6. Strengthening the spleen and removing phlegm and blood stasis demolition and party intervention in Bama miniature pig, compared with normal Bama miniature pigs, the expression of BACE1 in brain protein decreased significantly; 7.Immunohistochemistry showed that: compared with the normal group, there was significant difference between the content of HMGR in brain tissue of model group(P < 0.01); compared with the model group, diffent Chinese medicine prescription dismantlement and compound group effect on the HMGR content in the brain tissue had significant difference(P < 0.01); immunohistochemical staining showed increased in model group positive cells, and the drug group positive cells decreased, compared with the model group, the difference was statistically significant(P < 0.01) 8. Immunohistochemistry showed that: compared with the normal group, there was significant difference between the content of ABCA1 in brain tissue of model group(P < 0.01); compared with the model group, Chinese medicine prescription dismantlement and compound group effect on the ABCA1 content in the brain tissue had significant difference(P < 0.01); immunohistochemical staining showed increased in model group positive cells, and the drug group positive cells decreased, compared with the model group, the difference was statistically significant(P < 0.01);9. The results of immunohistochemical detection of CYP46 a showed that, compared with the normal group, model group, brain tissue increased CYP46 a content of each compound, and there is a significant difference(P < 0.01); compared with the model group, the increase of CYP46 a content in brain tissue of compound group, the difference was statistically significant(P < 0.01); compared with invigorating spleen and removing phlegm and blood stasis group, invigorating spleen group, reduce brain tissue group, dispelling phlegm and removing phlegm and blood stasis group of CYP46 a content, the difference was statistically significant(P < 0.01); compared with the group of invigorating spleen and dispelling phlegm, increase group, CYP46 a content of brain tissue and removing phlegm and blood stasis group, and the difference was statistically significant(P < 0.01); compared with the spleen and eliminating phlegm group, and removing phlegm and blood stasis group CYP46 decreased, the difference was not statistically significant.(P > 0.05). The expression of positive cells in the model group was increased by immunohistochemistry staining, and the expression of the positive cells in the drug group was decreased, compared with the model group(P < 0.01). 10. Compared with the normal group, model group, compound group brain tissue HMG coar protein expression increased significantly, and the difference is statistically significant(P < 0.01); compared with the model group, compound group HMGR protein expression increased significantly, and the difference is statistically significant(P < 0.01); compared with the spleen and eliminating phlegm and resolving blood stasis group, in the group of invigorating the spleen, dispelling phlegm group, eliminating phlegm and removing blood stasis group, cerebral tissue HMG coar protein expression decreased, the difference was statistically significant(P < 0.01)Compared with in the group of invigorating the spleen and dispelling phlegm group, eliminating phlegm and removing blood stasis group, HMG coar protein expression increased, the difference is statistically significant(P < 0.05); Jianpi Qutan group, eliminating phlegm and removing blood stasis group, compared to HMGR protein differentiallyexpressed no statistical significance(P > 0.05); 11. Western Blot the results showed that: compared with the normal group, model group, remove the HMGR protein group and invigorating the spleen and removing phlegm and blood stasis compound group of brain tissue expression had significant difference(P < 0.01); compared with the model group, Chinese medicine prescription dismantlement and compound group on the effect of HMGR protein expression were significantly different in the brain tissue(P < 0.01); compared with the spleen and removing phlegm and blood stasis group, eliminating phlegm, removing blood stasis and invigorating the spleen group group group there were significant differences on the HMGR expression in cerebral tissue; the group of invigorating spleen and removing blood stasis group, eliminating phlegm group, no significant difference on the expression of HMGR protein in brain tissue.(P > 0.05); 12. Western blot results show that compared with the normal group, model group, compound group brain tissue apo E protein expression increased and significant difference(P < 0.01); compared with the model group, compound group in the brain tissue of the APOE protein decreased expression of, the difference was statistically significant(P < 0.01); compared with invigorating the spleen and removing phlegm and blood stasis group, and removing phlegm and blood stasis group, spleen phlegm group and the group of invigorating spleen and brain tissue of apo E protein expression increased, there was significant difference(P < 0.01); in the group of invigorating the spleen, invigorating the spleen and removing phlegm group, eliminating phlegm and removing blood stasis group, brain tissue protein differentially expressed no statistical significance.(P > 0.05); 13. Western blot results show that compared with the normal group, model group, compound group brain tissue ABCA1 protein expression increased and significant difference(P < 0.01); compared with the model group, compound group in brain tissue of ABCA1 protein expression increased, the difference was statistically significant(P < 0.01); compared with the spleen and removing phlegm and bloodstasis group, Jianpi Qutan group, expectorant and removing blood stasis group and the group of invigorating spleen and brain tissue of ABCA1 protein expression decreased, the difference was statistically significant(P < 0.01); compared to in the group of invigorating the spleen and dispelling phlegm group, and removing phlegm and blood stasis group, brain ABCA1 protein expression had no significant difference.(P > 0.05).Conclusion:1. High fat diet can affect the blood lipid level and high fat diet for 1 week can cause dyslipidemia in coronary atherosclerosis model in pigs, with the extension of a high-fat diet, elevated concentrations of lipids in triglyceride(TG), total cholesterol(TC), low density lipoprotein(LDL), high density lipoprotein(HDL) concentration decreased. 2. High fat diet increased the peripheral blood lipid concentrations and peripheral blood lipid levels may influence Bama miniature pig brain amyloid beta levels change, increase in the brain of amyloid beta protein content and high fat diet 24 weeks can establish increased animal models of amyloid beta induced by high fat; 3. Jianpi Recipe, Jianpi Qutan recipe, eliminating phlegm and removing blood stasis, Jianpi Qutan Huayu Decoction can regulate the blood lipid level of the Bama miniature pig model, spleen and removing phlegm and Blood Stasis Formula better than spleen, invigorating spleen, eliminating phlegm and removing phlegm and blood stasis 4. Jianpi Recipe, Jianpi Qutan recipe, eliminating phlegm and removing blood stasis, Jianpi Qutan Huayu Decoction can reduce the deposition of amyloid beta in the brain, and reduce the expression of a beta. 5. Jianpi Recipe, Jianpi Qutan recipe, eliminating phlegm and removing blood stasis, Jianpi Qutan Huayu Decoction reduced amyloid beta protein content and production of one of the possible mechanisms is achieved by decreasing the expression of the beta amyloid metabolism key factor of BACE1 protein;6. The abnormal cerebral cholesterol metabolism can affect the A beta generation and metabolism 7. Jianpi Recipe, Jianpi Qutan recipe, eliminating phlegm and removing blood stasis, Jianpi Qutan Huayu Decoction can decrease the brain cholesterol synthesis and roll out the key enzyme of apo E protein, HMGR, ABCA1, and CYP46 protein expression was up-regulated, which effect of Jianpi Qutan Huayu decoction is superior to that of the spleen, invigorating the spleen and removing phlegm and phlegm stasis, invigorating spleen and removing phlegm and blood stasis 8. Jianpi Recipe, Jianpi Qutan recipe, eliminating phlegm and removing blood stasis, Jianpi Qutan Huayu Decoction intervention induced by high- fat diet a beta increased the possible mechanism of animal model is by down regulating the APOE protein expression and upregulated HMGR, ABCA1, and CYP46 protein expression to play a role. 9. Jianpi Recipe, Jianpi Qutan recipe, eliminating phlegm and removing blood stasis, Jianpi Qutan Huayu decoction may through peripheral regulation of Bama miniature pigs and blood lipid level in modulating Abeta generation secreted enzyme beta secretion enzyme activity and BACE1 protein expression and down regulate the expression of apo E protein and increase CYP46 a HMG coar and ABCA1 protein expression to intervene in the pathological process of Alzheimer’s disease in early stage beta amyloid formation, and in the early inhibition or delay the development of ad the occurrence and development. 10. Jianpi Recipe, Jianpi Qutan recipe, eliminating phlegm and removing blood stasis, Jianpi Qutan Huayu Decoction in regulating lipid, regulating A-beta generation and improve brain cholesterol metabolism effect is significant, the effect of Jianpi Qutan Huayu decoction is greater than that of Jianpi Qutan recipe, eliminating phlegm and removing blood stasis is larger than that of the spleen. Jianpi Qutan Huayu Decoction were excellent including invigorating spleen, removing blood stasis and removing phlegm.