| In recent years,with the gradual improvement of people’s living standards,the incidence of cerebrovascular diseases concerned based on atherosclerosis is increasing.According to the latest epidemiological research,atherosclerotic diseases have become one of the main causes of human death.In order to treat atherosclerotic related diseases,academia has been constantly exploring,but there is still no very effective treatment measures.We use "nei xiao ruan mai decoction" clinical atherosclerosis with obvious therapeutic effect,the patient blood lipid level and level of inflammatory factors have certain effect,speculated that the mechanism may be related to adenosine promote lipolysis UCP1 dependence on both A2A activation,to clear its curative effect and mechanism of action,to carry out "nei xiao ruan mai decoction" mechanism research and clinical observational studies.Methods:Trial 1(clinical trial):Screening of 50 cases diagnosed patients with carotid atherosclerotic plaques of outpatient service,"nei xiao ruan mai decoction" to give patients,and record the patient before taking the medicine,the IMT of carotid artery ultrasound report,after taking the longitudinal cutting thickness,transverse thickness and area of indicators,and according to the nature of age,gender,plaques for the statistical analysis of results,to evaluate "inside soft pulse elimination soup" efficacy of carotid atherosclerotic plaques.Experiment 2(basic experiment):According to different experimental requirements,ApoE knockout mice and C57BL6J mice were randomly divided into groups,and different doses of "nei xiao ruan mai decoction"were given.After the corresponding intervention conditions were completed,the samples to be tested were collected and their serum was stored in the refrigerator at-80 degrees Celsius.Biochemical methods were used to determine blood lipid levels and other related indicators.The tissue was stored in liquid nitrogen or formalin for fixation,and the morphology of aortic plaques in mice was observed by HE staining method to clarify the therapeutic effect,and the morphological characteristics of adipose tissue in mice were observed to clarify the activation of adipose tissue.Different kinds of primary antibodies were used to detect the expression of the target protein in the paraffin sections of adipose tissue of mice by immunohistochemical method.Western blot was used to detect the expression levels of target proteins such as UCP1 in adipose tissue of various parts,and the data were finally analyzed.Results:Trial 1(clinical trial):"The internal soft pulse soup" after treatment,patients with different gender,age and properties of different plaques the IMT of carotid artery atherosclerosis plaques,longitudinal cutting thickness,transverse thickness and plaques area are declining,IMT,longitudinal cutting thickness and plaques area decreased statistically difference,the thickness of crosscutting fall not statistically difference;Experiment 2(basic experiment):1.The effect of "neixiao ruanmai decoction" on atherosclerotic plaques and hs-crp in mice"Nei xiao ruan mai decoction":the area of atherosclerotic plaques in the intragastric group decreased significantly,and the hs-crp of both ApoE gene knockout mice and c57 mice decreased.The results of c57 mice were statistically different,while the results of ApoE gene knockout mice were not statistically different.Compared with the control group,the hs-crp of C57 mice decreased by 66.4%(*)in the low-dose group and 15.2%(*)in the high-dose group.2.Effect of "nei xiao ruan mai decoction" on Browning of mouse fat cells.According to the results of HE staining,the fat cells in the scapular area of ApoE-/-mice and C57 mice in the control group showed relatively full,large volume and single-compartment lipid droplets,and the staining was relatively shallow.The adipose cells of the mice in the "neixiao ruanmai decoction" group were smaller in size,darker in HE staining,more in nuclear staining,and more in number of adipose cells per unit visual field.The adipose tissue in the groin area of the mice in the gavage group also showed different degrees of activation,and the adipose tissue in the epididymis area had the least degree of activation.According to the results of immunohistochemical staining,compared with the control group,the positive staining of uncoupled protein 1(UCP1)in the group of "neixiao ruanmai decoction" was deeper than that in the control group.According to the results of Western Blot,compared with the control group,the expression of UCP1 protein in the adipose tissue of mice in the group of "neixiao ruanmai decoction" increased in a dose-dependent manner.3.Effect of "nei xiao ruan mai decoction" on serum lipid level in mice Neixiao ruanmai decoction showed no significant difference in serum lipid levels between ApoE knockout mice and C57 mice.4.Effect of "nei xiao ruan mai decoction" on adenosine A2A expression in adipose tissue of mice.According to the immunohistochemical staining results,the adenosine A2A positive staining of mice in the group of "neixiao ruanmai decoction" was deeper than that of the control group.According to the results of Western Blot,adenosine A2A protein expression in adipose tissue of mice in the group of "neixiao ruanmai decoction" increased with drug dose dependence compared with the control group.conclusionTrial 1(clinical trial):1."nei xiao ruan mai decoction" has the effect of reducing IMT,longitudinal thickness and area of carotid atherosclerotic plaque.2."nei xiao ruan mai decoction" reduced the IMT and longitudinal cutting thickness of plaques of female patients to a greater extent than that of male patients.3."nei xiao ruan mai decoction" is more effective for patients under the age of 65.4."nei xiao ruan mai decoction" has the largest IMT reduction for mixed plaques,the largest reduction for the longitudinal cutting thickness of soft plaques,and the largest reduction for the area of hard plaques.Experiment 2(basic experiment):1."nei xiao ruan mai decoction" can reduce the As plaque area of ApoE knockout mice.2."nei xiao ruan mai decoction" can reduce the level of hs-crp in mice.3."nei xiao ruan mai decoction" can activate UCP1 dependent lipolysis in mice,among which,it has no obvious effect on white adipose tissue in epididymis.4.There was no statistical difference in the effect of "neixiao ruanmai decoction" on the serum lipid level of mice.5."nei xiao ruan mai decoction" can activate adenosine A2A expression in the adipose tissue of mice,considering that its expression can promote UCP1 dependent lipolysis. |
PDF Full Text Request