Study on the mechanism of Qihuang Juyu Formula affecting the formation of atherosclerosis by regulating ApoE-/-mice macrophage related cytokines
Objective:To explore the mechanism of Qihuang Juyu Formula affecting atherosclerosis by regulating ApoE-/-mice macrophage related cytokines.Methods:Sixty-eight ApoE-/-mice were randomly divided into a model group(n=24),a traditional Chinese medicine group(n=22),and a control group(n=22),with 25 C57BU6 mice as the blank group.The atherosclerotic ApoE-/-mouse model was constructed by feeding high-fat diet.In the Chinese medicine group,Qihuang Juyu Formula Granules were administered to mice by gavage with 0.2 mL/20 g body weight.In the control group,atorvastatin calcium tablet suspension was administered to mice by gavage with 0.2 mL/20 g body weight.The blank group and model group were given an equal amount of 0.9％sodium chloride solution by gavage.All were continuously administered for 12 weeks.Observe the general state and body mass of each group of mice.Aortic HE staining,Masson staining,and oil red O staining were used to observe the pathological morphology and plaque area of the blood vessels.Serum ELISA detection of macrophage related factors expression,including interleukin(IL)-1β,IL-4,IL-13,interferon-γ(IFN-γ),Lipopolysaccharides(LPS),transforming growth factors-β(TGF-β),IL-6,IL-10,IL-12.Immunofluorescence was used to observe the expression of macrophage surface markers iNOS and CD206.Results:The mice in the model group,traditional Chinese medicine group,and control group showed signs of mental fatigue,dull hair color,rapid growth in the early stage,and gradually slowing down later.The quality of the model group was significantly higher than that of other groups(P＜0.01).The pathological images of HE staining,Masson staining,and Oil Red O staining showed that the model group had a significant increase in plaque area,a significant decrease in collagen fiber content,and a significant increase in aortic red stained lipids compared to the blank group(P＜0.01).The traditional Chinese medicine group and control group had a significant decrease in plaque area,an increase in collagen fiber content,and a significant decrease in aortic red stained lipids compared to the model group(P＜0.01).The ELISA test results show that the serum IL-1β,IFN-γ,LPS,IL-6,IL-12 of the model group mice were significantly higher than the blank group(P＜0.05),IL-4,IL-13,TGF-β,IL-10 was significantly lower than that of the blank group(P＜0.05);Comparison between traditional Chinese medicine group and model group,mouse serum IL-1β,IFN-γ,LPS,IL-6,and IL-12 were significantly reduced(P＜0.05),while the expression levels of IL-4,IL-13,TGF-β,and IL-10 were significantly increased(P＜0.05).Immunofluorescence staining showed that the positive expression of M1 type macrophage marker iNOS was higher in the model group(P＜0.01),and lower in the traditional Chinese medicine group(P＜0.05).The positive expression of M2 type macrophage marker CD206 was lower in the model group than in the traditional Chinese medicine group(P＜0.05).Conclusion:Qihuang Juyu Formula can reduce M1 type macrophages and inhibit the expression of M1 type macrophage related factors IL-1β,IFN-γ,LPS,IL-6,IL-12,and increase M2 macrophages,promote the generation of M2 macrophage related factors IL-4,IL-13,TGF-β,and IL-10,this may be the mechanism of its intervention in the occurrence and development of atherosclerosis.
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