To study the effects of moxibustion method of “warming Yang and dredging vessels” on the morphology and function of mitochondria and the level of apoptosis in ApoE-/- atherosclerosis(AS)mice, and to explore the mechanism of prevention and treatment of AS.
Twenty-two 8-week-old SPF-grade male ApoE-/- mice were randomly allocated into the model group and the moxibustion group, each comprising 11 mice, and the mice were placed on a high-fat diet to establish the atherosclerosis model. Eleven C57BL/6J mice formed the control group and were administered the standard chow. The intervention commenced on the initial day of the modeling for each group, with only the control and model groups being restricted, while the mice in the moxibustion group were positioned at “Danzhong(RN17)”, “Shenque(RN8)” and “Neiguan(PC6)”(bilaterally). The mice in the moxibustion group received moxibustion at “Danzhong(RN17)”, “Shenque(RN8)”, “Neiguan(PC6)”(bilaterally)and “Xuehai(SP10)”(bilaterally), administered daily for 30 minutes, five times per week. The samples were collected 12 weeks after intervention. The levels of serum triglyceride(TG)and total cholesterol(TC)were quantified by using an enzymatic assay, while the levels of serum low-density lipoprotein cholesterol(LDL-C)and high-density lipoprotein cholesterol(HDL-C)were assessed by a colorimetric test. The utilization of hematoxylin-eosin(HE)staining facilitated the examination of the pathological morphology and the structure of the thoracic aorta in mice. The transmission electron microscopy was utilized to analyze the morphology of mitochondria in the thoracic aorta, and the whilst flow cytometry(JC-1)was used to evaluate the mitochondrial membrane potential in the thoracic aorta. The changes in mitochondrial membrane potential(MMP)were assessed by using JC-1. The cytochrome C(Cyt C)protein expression in the thoracic aorta was examined through immunofluorescence staining and the protein expression levels of Bax, Bcl-2 and caspase-3 in the thoracic aorta were measured by using immunohistochemistry and Western blot analysis.
Compared with those of the control group, the levels of serum TC, TG and LDL-C of mice in the model group were significantly higher(P < 0.01)and HDL-C levels were significantly lower(P < 0.01). The thoracic aortic structures in mice showed intimal thickening, endothelial cell degeneration, swelling and detachment, and the foam cells were seen. The mitochondria were massively swollen and the number of cristae was greatly reduced. Mitochondrial membrane potential levels were significantly reduced(P < 0.01). The Cyt C protein levels were significantly higher in thoracic aortic tissue(P < 0.01). The protein expression levels of Bax and Caspase-3 were significantly higher(P < 0.01), and the protein expression level of Bcl-2 was significantly lower(P < 0.01).
“Warming Yang and dredging vessels” moxibustion prevents and treats AS by improving the morphology and function of mitochondria and reducing the level of apoptosis.
| 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 |