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Objective To investigate application value of three-dimensional light sheet microscopy imaging for evaluating intraplaque neovascularization in arterial plaques and the efficacy of intervention, and to assess the effect of melatonin (MLT) on neovascularization by these means. Methods Thirty-six ApoE-/- model mice were randomly divided into three groups (n=12): vehicle group, MLT group, and MLT+GW9662 intervention group (MLT+GW). The mice were treated with vehicle, MLT alone, or MLT combined with peroxisome proliferator activated receptor‑γ (PPARγ) inhibitor GW9662, respectively. The carotid arteries of the models were three-dimensionally imaged using a light sheet microscopy, and the length, volume and other indicators of neovascularization were quantitatively analyzed using Imaris software. Subsequently, CD31 immunohistochemical staining was performed for verification. Results The light sheet microscopy preliminarily achieved the three-dimensional visualization of intraplaque neovascularization, and its structure was observed to be three-dimensionally reticular and scattered. The results of Imaris quantitative analysis showed that, compared with vehicle group, the total intraplaque neovas cularization length in the MLT group was shortened [(15.79±12.90) mm vs. (33.42±11.16) mm, P<0.05], the total volume was reduced [(1.34±1.47)×10-3 mm3 vs. (13.44±7.35)×10-3 mm3, P<0.05], and the volume ratio was decreased (0.44%±0.47% vs. 3.76%±1.74%, P<0.05). The above indicators in MLT+GW group were significantly increased compared with those in MLT group [total length: (35.31±4.69) mm, total volume: (8.87±3.46)×10-3 mm3, volume ratio: 2.89%±0.38%; P<0.05]. The CD31 immunohistochemical staining also supported the above findings (P<0.05). Conclusions Based on the light sheet microscopy imaging technology, the three-dimensional visualization and quantitative analysis of intraplaque neovascularization were preliminarily realized. It was found that MLT could reduce the overall burden of intraplaque neovascularization, and PPARγ might be involved in its regulatory process.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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