Autophagosome activity in macrophage for atherosclerotic plaques in ApoE−/− mice enhanced by Tiaozhi Tongmai Granules

Abstract

Objective: Atherosclerosis is the fundamental pathophysiologic component of cardiovascular disease, and Tiaozhi Tongmai Granules show great efficiency in the treatment of the disease. However, the mechanism of Tiaozhi Tongmai Granules is still unclear. In this study, we have combined experiments with network pharmacology to explore the anti-atherosclerosis mechanism of Tiaozhi Tongmai Granules. Method: 120 male ApoE−/− mice were randomly divided into three groups: the model group, Chinese herb group and Atorvastatin group. The model group, Atorvastatin group and Chinese herb group were fed with a high-fat diet, a high-fat diet plus atorvastatin (5.1 mg/kg/d) and a high-fat diet plus Tiaozhi Tongmai Granules (16.5 g/kg/d) for 16 weeks, respectively. Atherogenesis was identified by H&E staining. The colocalization of neutral lipid stain BODIPY and microtubule-associated protein 1 light chain 3 (LC3) and the colocalization of BODIPY and lysosomal-associated membrane protein 1 (LAMP1) within ApoE−/− mice aortic plaques were tested using fluorescence confocal microscopy and the Pearson's coefficients were calculated. To further explore the anti-atherosclerosis mechanism of Tiaozhi Tongmai Granules, the network pharmacology was used to construct the herb-compound-target network. Results: The size of the aortic lipid plaque in the Chinese herb group and Atorvastatin group were smaller compared with the model group on the 16th week. Compared with the model group on the 16th week, the BODIPY and LC3 colocalization rate, the BODIPY and LAMP1 colocalization rate of the Chinese herb group and Atorvastatin group all presented significant increase in the aortic plaque (P < .001), showing that Tiaozhi Tongmai Granules could enhance autophagosome activity in the macrophage. In the herb-compound-target network, 17 active compounds and 27 targets were obtained through literature searching and using LHRI & DAVID Bioinformatics. It was found that 23 targets were correlated with the macrophage. Some of them participated in macrophage inflammatory response, and the other targets could promote/inhibit phagocytosis of the macrophage. It was hypothesized that the active compounds of Tiaozhi Tongmai Granules were acting on these targets and having y the biological effects. Conclusions: In the progression stage of atherosclerosis, Tiaozhi Tongmai Granules can still make the macrophage have higher autophagosome activity, and play a role of anti-atherogenesis.

Keywords

• Atherosclerosis
• Autophagy
• Chinese medicine
• Immunofluorescence
• System pharmacology