| Purpose:According to the theory of"treating from spleen",the experimental study of Jianpi Qutan Fang regulating macrophage polarization and anti-Apo E-/-atherosclerosis in mice was discussed.This study selected high-fat diet combined with single cage feeding to establish spleen deficiency phlegm turbidization AS model,and used the method of invigorating qi to strengthen spleen and removing phlegm and removing blood stasis to intervene,providing theoretical and experimental basis for the treatment of atherosclerosis.Materials and methods:Eighteen Apo E-/-mice were randomly divided into model group,atorvastatin group and Jianpi Qutanfang group,with 8 mice in each group.Six C57BL/6J mice were selected as the normal group,and the claws were numbered after 1 week of adaptive feeding.From the second week,normal group was fed with ordinary diet,and the other three groups were fed with high-fat diet.Atorvastatin group was given atorvastatin2.4mg/（kg·d）gavage,Jianpi Qutanfang group was given Jianpi Qutanfang 11.88g/（kg·d）gavage,normal group and model group were given equal volume normal saline gavage,modeling and administration were performed simultaneously.After 12 weeks,spleen deficiency and phlegm turbid AS model was evaluated.During the experiment,the body weight and stool characteristics of mice were observed every week.At the 12th week,after modeling and administration,the animal behavior was evaluated by open field experiment and treadmill experiment.Body composition detection;The pathological sections of aorta in different groups of mice were stained with HE to observe the plaque formation.Serum levels of tumor necrosis factor-α（TNF-α）,interleukin-6（IL-6）and IL-10 were determined by enzyme-linked immunosorbent assay（ELISA）.The expression levels of macrophage polarization markers nitric oxide synthase（i NOS）in M1 macrophages and argininase-1（Arg-1）in M2 macrophages were detected by immunoblotting（WB）.Serum total cholesterol（TC）,low density lipoprotein（LDL）,triglyceride（TG）,high density lipoprotein（HDL）levels of mice were detected by biochemical analyzer.Results:1.General status:After modeling,mice in the model group fed with high fat diet showed dull hair color and fur shedding,decreased mobility,loose stools and loose stools.Compared with model group,activity,fecal characters and hair color of mice in Jianpi Qutan Fang group and atorvastatin group were improved.2.Body weight:Compared with normal group,body weight of mice in model group increased significantly（P<0.01）,and the difference was significant.The body weight of mice in atorvastatin group and Jianpi Qutanfang group was significantly lower than that in model group（P<0.01）,and the difference was statistically significant.3.Body composition content:compared with normal group,body fat in model group was significantly decreased（P<0.01）,and the difference was significant.The body fat of atorvastatin group was significantly higher than that of model group（P<0.01）,and the difference was statistically significant.The body fat of Jianpi Qutanfang group was increased compared with model group（P>0.05）,but the difference was not statistically significant.4.Fecal score:The fecal BSFS score of the 4 groups of mice was analyzed by one-way variance analysis.At 6 weeks,compared with the normal group,fecal score of the model group was significantly higher（P<0.01）,and there was a significant difference.Compared with model group,fecal scores of mice in Jianpi Qutanfang group and atorvastatin group were increased（P>0.05）,and the difference was not statistically significant.At week 12,fecal score of model group was significantly higher than that of normal group（P<0.01）,and the difference was statistically significant.The fecal score of Jianpi Qutan formula group was significantly lower than that of model group（P<0.01）,and the difference was statistically significant.The results indicated that Jianpi Qutan formula can obviously reduce the stool score of mice with spleen deficiency and phlegm turbid AS,and can improve the symptoms of spleen deficiency and phlegm turbid loose stool in mice with AS.5.Open field experiment:Compared with normal group,the total distance of mice in model group was significantly shortened（P<0.01）,Compared with model group,the exercise distance of mice in Jianpi Qutanfang group and atorvastatin group had an increasing trend（P>0.05）.6.Treadmill experiment:Compared with normal group,running exhaustion time of mice in model group was significantly decreased（P<0.01）.The running exhaustion time of Jianpi Qutanfang group was significantly longer than that of model group（P<0.01）,and the difference was statistically significant.Compared with model group,the running exhaustion time of atorvastatin group had no significant difference（P>0.05）.The results showed that Jianpi expectorant can obviously increase the running exhaustion time of mice with spleen deficiency and phlegm turbid AS,and improve the symptoms of spleen deficiency,fatigue and fatigue of mice with AS.7.HE staining:Compared with the normal group,the structure of aorta in the model group was disorganized,the tube wall was thickened,foam cells were visible in the medium membrane,and obvious atherosclerotic plaque was formed.Compared with model group,the aorta structure of Jianpi Qutanfang group and atorvastatin group was normal,the inner membrane was smooth,the tube wall was not thickened,and no plaque formation was observed.The results showed that the model of mice with spleen deficiency and phlegm turbidities was successful,and Jianpi Qutan prescription had obvious improvement effect on AS.8.Blood lipid detection:Compared with normal group,the contents of TC,TG and LDL in serum of model group were significantly increased（P<0.01）,while the content of HDL was significantly decreased（P<0.01）.Compared with model group,the contents of TC,TG and LDL in serum of Jianpi Qutanfang group and atorvastatin group were significantly decreased（P<0.01）,while the content of HDL was significantly increased（P<0.01）,indicating that Jianpi Qutanfang can improve the blood lipid of mice with spleen deficiency and phlegm turbidities.9.Inflammatory factor detection:The contents of IL-6 and TNF-αin serum of model group were significantly increased compared with normal group（P<0.01）,and the content of IL-10was significantly decreased compared with normal group（P<0.01）.Compared with model group,the contents of IL-6 and TNF-αin serum of Jianpi Qutanfang group and atorvastatin group were significantly decreased（P<0.01）,and the content of IL-10 in serum of Jianpi Qutanfang group was significantly increased（P<0.01）,indicating that Jianpi Qutanfang could reduce the level of pro-inflammatory factors and promote the level of anti-inflammatory factors.Thus,the inflammation level of AS mice was reduced.10.Detection of macrophage polarization markers:Compared with normal group,i NOS Arg-1protein expression in aorta tissue of model group was significantly increased（P<0.01）.Compared with model group,i NOS Arg-1 protein expression in aorta tissues of mice in Jianpi Qutanfang group and atorvastatin group was significantly decreased（P<0.01）.The results indicated that Jianpi Qutan Fang could reduce the M1-type polarization of macrophages,and enhance the stability of plaque.Conclusion:Jianpiqutanfang has an intervention effect on the formation of atherosclastic plaques in the aorta endometrium of mice with spleen deficiency and phlegm turbidness,and can improve loose stool symptoms,increase the running time and activity of mice,improve blood lipid level,reduce the expression of pro-inflammatory factors and increase the expression of anti-inflammatory factors,promote the polarization of macrophages in aortic plaques to M1 type,and reduce the damage caused by inflammation to blood vessels.It can increase the stability of plaque and inhibit the occurrence and development of AS,which provides experimental basis for the treatment of atherosclerosis by invigorating qi and strengthening spleen and removing phlegm and removing blood stasis. |
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