| Purpose:To investigate the effects of Tongmai Jiang Zhuo Decoction on chronic psychological stress atherosclerosis mice and foam cells in vitro by animal experiments.The levels of lipid,liver index,corticosterone,aortic lesion area,SREBP-1 protein and SREBP-2protein expression and in vitro were observed.The effects of total cholesterol,free cholesterol content,cholesterol efflux and the expression of ABCA1,miR-33 a,miR-33 b protein and mRNA in foam cells were investigated,so as to explore the mechanism of Tongmai Jiang Zhuo Decoction in treating chronic psychological stress atherosclerosis and the mechanism of its prevention and treatment of atherosclerosis,so as to provide experimental basis and theory for clinical treatment of chronic psychological stress atherosclerosis.Basis.Materials and methods: Experiment 1.Male 4-week-old ApoE-/-mice were randomly divided into 5 groups,28 in each group,They were control group?normal group,a group?,as model group?atherosclerosis model group,B group?,as model stress group?atherosclerosis model + chronic psychological stress model,C group?,as model stress Chinese medicine treatment group?atherosclerosis model + chronic psychological stress model + Tongmai Jiangzhuo Decoction group,D group?,as model stress positive treatment group?atherosclerosis model?+Chronic psychological stress model + simvastatin +loraz EPam group,E group),through the observation of the control group,AS model group,AS model stress group mice general situation,behavioral results,corticosterone content,serum and liver tissue lipid levels and aortic lesion area,the establishment of apoE-/-mouse atherosclerosis model of chronic psychological stress was evaluated.By observing the liver index,the content of corticosterone,the level of lipid in serum and liver tissue,the area of aortic lesions,the content of SREBP-1 protein and SREBP-2 protein in aorta and liver,the mechanism of Tongmai Jiangzhuo Decoction in the treatment of chronic psychological stress atherosclerosis was explored.Experiment two,THP-1 cells with 2×106/ml concentration were divided into 4 groups: control group?blank group?,model group?ox-LDL treatment group?,model treatment group?ox-LDL+ beta cyclodextrin treatment group?,model TDMStreatment group?ox-LDL+TDMS treatment group?,total cholesterol,free cholesterol content,cholesterol efflux rate,ABCA1,miR-33 a and miR-33 b eggs in each group were observed.To explore the mechanism of Tongmai Jiangzhuo Decoction in regulating blood lipid metabolism and preventing as.Result:1 Establishment and evaluation of atherosclerosis model of ApoE-/-mice under chronic psychological stress1.1 comparison of weight results?1?There was no significant difference in body weight between groups?P > 0.05?.?2?Compared with the control group,there was no difference in as model group?P > 0.05?,as model stress group and as model stress treatment group significantly reduced?P < 0.01?;compared with as model group,as model stress group and as model stress treatment group significantly reduced?P < 0.01?,and there was no difference between the two groups?P >0.05?.?3?Compared with the control group,there was no difference in as model group?P >0.05?,as model stress group and as model stress treatment group significantly reduced?P<0.01?;compared with as model group,as model stress group and as model stress treatment group significantly reduced?P < 0.01?,and there was no difference between the two groups?P > 0.05?.?4?Compared with the control group,as model group increased significantly?P <0.01?,as model stress group decreased?P < 0.05?,as model stress treatment group had no difference?P > 0.05?;compared with as model group,each group decreased significantly?P <0.01?;compared with as model stress group,as model stress treatment group increased?P <0.05?.1.2 comparison of food consumption results?1?Results: compared with the control group,there was no difference in as model group?P > 0.05?,as model stress group and as model stress treatment group?P < 0.05?;compared with as model group,as model stress group and as model stress treatment group were significantly reduced?P < 0.01?,and there was no difference between the two groups?P >0.05?.?2?Results: compared with the control group,as model group increased significantly?P< 0.01?,as model stress group and as model stress treatment group were significantly less?P<0.01?;compared with as model group,as model stress group and as model stress treatment group decreased significantly?P < 0.01?,and there was no difference between the two groups?P > 0.05?.?3?Results: compared with the control group,as model group increased significantly?P < 0.01?,as model stress group decreased significantly?P < 0.01?,as model stress treatment group had no difference?P > 0.05?;compared with as model group,each group decreased significantly?P < 0.01?;compared with as model stress group,as model stress treatment group increased?P < 0.01?.1.3 comparison of sweet water preference experiment results?1?Compared with the control group,there was no difference in as model group?P >0.05?,as model stress group and as model stress treatment group?P < 0.01?;compared with as model group,as model stress group and as model stress treatment group were significantly reduced?P < 0.01?,and there was no difference between the two groups?P > 0.05?.?2?Compared with the control group,as model group,as model stress group and as model stress treatment group were significantly reduced?P < 0.01?;compared with as model group,as model stress group and as model stress treatment group were significantly reduced?P < 0.01?,and there was no difference between the two groups?P > 0.05?.?3?Compared with the control group,the as model group and the as model stress group were significantly reduced?P < 0.01?,and there was no difference?P > 0.05?between the as model stress group and the as model stress group;compared with the as model group,the as model stress group was significantly reduced?P < 0.01?,and the as model stress group was significantly increased?P < 0.01?;compared with the as model stress group,the as model stress group was significantly reduced?P < 0.01?Significantly increased?P < 0.01?.1.4 comparison of open field test results1.4.1 number of grid crossing within 5min?1?Compared with the control group,there was no difference in as model group?P >0.05?,as model stress group and as model stress treatment group?P < 0.05?;compared with as model group,as model stress group and as model stress treatment group decreased?P < 0.05?,and there was no difference between the two groups?P > 0.05?.?2?Compared with the control group,as model group,as model stress group and as model stress treatment group were significantly reduced?P < 0.01?;compared with as model group,as model stress group and as model stress treatment group were significantly reduced?P < 0.01?,and there was no difference between the two groups?P > 0.05?.?3?Compared with the control group,as model group and as model stress group decreased significantly?P < 0.01?,as model stress group had no difference?P > 0.05?;compared with as model group,as model stress group decreased significantly?P < 0.01?,as model stress group increased significantly?P < 0.01?;compared with as model stress group,as model stress group,as model stress group Significantly increased?P < 0.01?.1.4.2 total standing time within 5min?1?Compared with the control group,there was no difference in the total standing time within 5 minutes at the end of 4 weeks in as model group?P> 0.05?,and there was a significant increase in as model stress group and as model stress treatment group?P< 0.01?;compared with as model group,there was a significant increase in as model stress group and as model stress treatment group?P < 0.01?,and there was no difference between the two groups?P> 0.05?.?2?Compared with the control group,there was no difference in the total standing time within 5 minutes at the end of 8 weeks?P> 0.05?,and there was a significant increase in as model stress group and as model stress treatment group?P< 0.01?;compared with as model group,there was a significant increase in as model stress group and as model stress treatment group?P < 0.01?,and there was no difference between the two groups?P >0.05?.?3?Compared with the control group,the total standing time of as model group decreased significantly?P < 0.01?,the stress group increased significantly?P < 0.01?,and there was no difference in as model stress treatment group?P > 0.05?;compared with as model group,as model stress group and as model stress treatment group increased significantly?P < 0.01?;compared with as model stress group,as model stress treatment group decreased significantly?P < 0.01??1.5 comparison of plasma corticosterone content?1?At the end of 4 weeks,there was no difference between as model group and control group?P > 0.05?,and as model stress group was significantly higher?P < 0.01?;compared with as model group,as model stress group was significantly higher?P < 0.01?.?2?At the end of 8 weeks,compared with the control group,as model group and as model stress group increased significantly?P< 0.01?;compared with as model group,as model stress group increased significantly?P < 0.01?.?3?At the end of 12 weeks,compared with the control group,as model group and as model stress group increased significantly?P < 0.01?;compared with as model group,as model stress group increased significantly?P < 0.01?.1.6 liver index comparison results?1?At the end of 4 weeks,there was no difference between as model group and control group?P > 0.05?,and as model stress group was significantly higher?P < 0.01?;compared with as model group,as model stress group was significantly higher?P < 0.01?.?2?At the end of 8 weeks,compared with the control group,as model group and as model stress group increased significantly?P < 0.01?;compared with as model group,as model stress group increased significantly?P < 0.01?.?3?At the end of 12 weeks,compared with the control group,as model group and as model stress group increased significantly?P < 0.01?;compared with as model group,as model stress group increased significantly?P < 0.01?.1.7 comparison of serum lipid and liver lipidThe results of serum and liver of TC,TG,LDL-C were compared:?1?at the end of 4weeks,there was no difference between as model group and control group?P > 0.05?,and as model stress group was significantly higher?P < 0.01?;compared with as model group,as model stress group was significantly higher?P < 0.01?.?2?At the end of 8 weeks,compared with the control group,as model group and as model stress group increased significantly?P <0.01?;compared with as model group,as model stress group increased significantly?P < 0.01?.?3?At the end of 12 weeks,compared with the control group,as model group and as model stress group increased significantly?P < 0.01?;compared with as model group,as model stress group increased significantly?P< 0.01?.HDL-C serum results:?1?compared with the control group at the end of 4 weeks,there was no difference in as model group?P > 0.05?,as model stress group was significantly lower?P <0.01?;compared with as model group,as model stress group was significantly lower?P <0.01?.?2?At the end of 8 weeks,compared with the control group,as model group and as model stress group decreased significantly?P < 0.01?;compared with as model group,as model stress group decreased significantly?P < 0.01?.?3?At the end of 12 weeks,compared with the control group,as model group and as model stress group decreased significantly?P <0.01?;compared with as model group,as model stress group decreased significantly?P <0.01?.1.8 comparison of aortic lesion area resultsAt the end of 12 weeks,compared with the control group,as model group and as model stress group increased the area of aortic lesions?P < 0.01?;compared with as model group,as model stress group increased the area of aortic lesions?P < 0.01?.2 The experimental results of Tongmai Jiangzhuo Decoction on the treatment of ApoE-/-mouse as model of chronic psychological stress based on SREBPs pathway regulation of SREBP-1 and SREBP-22.1 comparison of liver index resultsCompared with the control group,each group was significantly higher?P < 0.01?;compared with the as model group,the as model stress group was significantly higher?P <0.01?,the as model stress treatment group and the as model stress control group were significantly lower?P < 0.01?;compared with the as model stress group,the as model stress treatment group and the as model stress control group were significantly lower?P < 0.01?,and there was no difference between the groups?P > 0.05?.2.2 comparison of total cholesterol resultsCompared with the control group,the results of TC in serum and liver tissue were significantly higher in each group?P < 0.01?;compared with the as model group,the as model stress group was significantly higher?P < 0.01?,the as model stress treatment group and the as model stress control group were significantly lower?P < 0.01?;compared with theas model stress group,the as model stress treatment group and the as model stress control group were significantly lower?P < 0.01?,There was no difference between the two groups?P > 0.05?.2.3 comparison of triglyceride resultsCompared with the control group,TG in serum and liver tissue increased significantly?P< 0.01?;compared with the as model group,the as model stress group increased significantly?P < 0.01?,the as model stress treatment group and the as model stress control group decreased significantly?P < 0.01?;compared with the as model stress group,the as model stress treatment group and the as model stress control group decreased significantly?P< 0.01?,There was no difference between the two groups?P > 0.05?.2.4 comparison of HDL-C resultsCompared with the control group,HDL-C in serum and liver tissue decreased significantly?P < 0.01?;compared with the as model group,the as model stress group decreased significantly?P < 0.01?,the as model stress treatment group and the as model stress control group increased significantly?P < 0.01?;compared with the as model stress group,the as model stress treatment group and the as model stress control group increased significantly?P < 0.01?,There was no difference between the two groups?P > 0.05?.2.5 comparison of LDL-C resultsCompared with the control group,LDL-C in serum and liver tissue increased significantly?P < 0.01?;compared with the as model group,the as model stress group increased significantly?P < 0.01?,the as model stress treatment group and the as model stress control group decreased significantly?P < 0.01?;compared with the as model stress group,the as model stress treatment group and the as model stress control group decreased significantly?P < 0.01?,There was no difference between the two groups?P > 0.05?.2.6 comparison of corticosterone resultsCompared with the control group,the content of corticosterone in each group was significantly higher?P < 0.01?;compared with the as model group,the stress group of as model was significantly higher?P < 0.01?,the stress treatment group of as model and thestress control group of as model were lower?P < 0.05?;compared with the stress group of as model,the stress treatment group of as model and the stress control group of as model were significantly lower?P < 0.01?,and there was no difference between the groups?P>0.05??2.7 comparison of aortic lesion area resultsCompared with the control group,the area of aortic lesions in each group increased significantly?P < 0.01?;compared with the as model group,the as model stress group increased significantly?P < 0.01?,the as model stress treatment group and the as model stress control group decreased?P < 0.05?;compared with the as model stress group,the as model stress treatment group and the as model stress control group decreased significantly?P < 0.01?,and there was no significant difference between the groups The difference?P > 0.05?.2.8 comparison of protein contents of SREBP-1 and SREBP-2Compared with the control group,the contents of SREBP-1 and SREBP-2 in the aorta increased significantly?P < 0.01?;compared with the as model group,the as model stress group increased significantly?P < 0.01?,the as model stress treatment group and the as model stress control group decreased?P < 0.05?;compared with the as model stress group,the as model stress treatment group and the as model stress control group decreased significantly?P< 0.05?There was no difference between the two groups?P > 0.05?.Compared with the control group,the content of SREBP-1 and SREBP-2 in liver tissue increased significantly in other groups?P < 0.01?;compared with the as model group,the stress group of as model increased significantly,the stress treatment group and the stress control group of as model decreased significantly?P < 0.05?;compared with the stress group of as model,the stress treatment group and the stress control group of as model decreased significantly?P < 0.01?,There was no difference between the two groups?P > 0.05?.3Tongmai Jiangzhuo Decoction Based on SREBPs Signaling Pathway Regulates Cholesterol Outflow from THP-1 Macrophage-derived Foam Cells3.1 TDMS effects on cell growthCompared with the inhibition od value of cell proliferation,compared with THP-1+PMA,there was no difference in T+P+normal serum group?P>0.05?,TP+5% drug-containing group,T+P+10% drug-containing group,T+P+15% drug-containing group,T+P+20%drug-containing group decreased?P<0.05?;compared with T+ P+normal serum group,T+P+5% drug-containing group,T+P+ 10% Drug-containing group,T+P+15%drug-containing group,T+P+20% drug-containing group decreased?P<0.05?;compared with T+P+5% drug-containing group,T+P+10% drug-containing group There was no difference in the drug group?P>0.05?,the T+P+15% drug-containing group and T+P+20% drug-containing group decreased?P<0.05?,and there was no difference between the two groups?P>0.05?3.2 Identification of foam cellsAfter PMA-induced differentiation,THP-1 cells were observed under microscope to change from suspension growth to adherent growth,and the cells grew pseudopods.Oil red O staining showed that the macrophages before ox-LDL induction had no staining positive cells,while the intracellular lipid accumulation of macrophages induced by ox-LDL was positive for oil red O staining,indicating that the foam was induced Cell success.3.3 Total and free cholesterol content and cholesterol efflux rate in cellsCompared with the control group,the total cholesterol and free cholesterol in the model group?ox-LDL treated group?increased?P<0.05?.Compared with the model group?ox-LDL treatment group?,the total cholesterol and free cholesterol contents in the model treatment group?ox-LDL + ?-cyclodextrin treatment group?and the model TDMS group?ox-LDL+TDMS treatment group?were both Decrease?P <0.05?.Compared with the control group,the cholesterol efflux rate of the model group decreased?P <0.05?;compared with the model group,the cholesterol efflux rate of the model treatment group and the TDMS group increased?P <0.05?;and there was no statistical difference between the two groups.3.4 Effect of Tongmai Jiangzhuo Decoction-containing serum on the translational levels of foam cells ABCA1Compared with the control group,the expression level of ABCA1 protein in the model group decreased?P <0.05?.Compared with the model group,the ABCA1 protein expression level in the model treatment group and the model TDMS group increased?P <0.05?.3.5 Effect of Tongmai Jiangzhuo Decoction-containing serum on the transcription levels of ABCA1,miR-33 a,miR-33 b in foam cellsCompared with the control group,the levels of ABCA1 mRNA in the model group decreased?P <0.05?,and the expression levels of miR-33 a and miR-33 b mRNA increased?P<0.05?.Compared with the model group,the levels of ABCA1 mRNA expression in the model treatment group and the model TDMS group were increased?P <0.05?,and the levels of miR-33 a and miR-33 b mRNA were decreased?P <0.05?.Conclusion:1 ApoE-/-mice with high-fat diet combined with chronic psychological stress can establish an animal model of chronic psychological stress atherosclerosis.2 Tongmai Jiangzhuo Decoction can reduce liver index,corticosterone content and lipid levels in the body,down-regulate SREBP-2,SREBP-1 protein expression,reduce aortic disease and treat chronic psychological stress atherosclerosis.3 Tongmai Jiangzhuo Decoction-containing serum can regulate the expression of ABCA1,miR-33 a,miR-33 b through the SREBPs signal pathway,thereby increasing the cholesterol efflux rate,reducing the cholesterol content in foam cells,and achieving the purpose of preventing and curing atherosclerosis. |
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