| OBJECTIVE:To investigate the effects of different frequencies of mild moxibustion on lipid metabolism related proteins in rabbits with atherosclerosis(AS).Methods:Fifty SPF rabbits were numbered after one week of adaptive feeding.They were randomly divided into normal groups(13)and model groups(37).The rabbits were modeled using a high-fat diet plus immune injury to replicate the atherosclerosis model.After the model was successfully established,36 rabbits were randomly divided into 12 model groups,12 low-frequency moxibustion groups and 12 high-frequency moxibustion groups.During the moxibustion intervention,the normal group and the model group were given ordinary feed,grabbed and bundled,and were not given moxibustion intervention.Rabbits in the low frequency and high frequency moxibustion groups were given ordinary feed and grabbed and bundled.Gentle moxibustion is used to treat atherosclerosis in acupoints of Danzhong,Shenque,Neiguan(double)and Xuehai(double).The high frequency group was treated once a day,and the intervention group was treated once every other day for 30 minutes each time for a total of 4 weeks.Observe the general conditions,appetite and stool changes of rabbits in each group during moxibustion intervention.Measure serum total TC,TG,HDL-C and LDL-C after the intervention and calculate TC/HDL-C,LDL-C/HDL-C ratio.Observation of pathological changes and ultrastructure of aortic atherosclerosis with light microscope and electron microscope.Western blot was used to detect the expression of AMPKa,LXRa and ABCA1 protein in liver and aorta.and Real time PCR was used to detect AMPKα,LXRα and ABCA1 mRNA expression in liver and aorta.Research result:1.General situation of rabbits in each groupIn the process of modeling and moxibustion intervention,rabbits in each group had smooth hair,good vitality and good overall condition.There was no significant difference in appetite between rabbits in each group.In terms of animal excrement,the rabbits in the model group had feces without forming stools.Except for occasional diarrhea due to weather changes,the molds of the other three groups were normal.2.Changes in blood lipid levels and ratios of rabbits in each group(1)Changes in serum TG content:The TG content of the rabbits in the model group was higher than that in the normal group,and there was a significant statistical difference compared with the rabbits in the normal group(P<0.01);the serum TG content of the rabbits in the high and low frequency groups was significantly reduced Compared with the model group,it was statistically significant(P<0.01);the serum TG content in the high frequency group was not statistically significant compared with the low frequency group(P>0.05).(2)Changes in serum TC content:The TC content of rabbits in the model group was significantly higher than that in the normal group,and there was a significant statistical difference compared with the rabbits in the normal group(P<0.01);the serum TC contents of the rabbits in the high and low frequency groups were significant Decreased,compared with the model group,it has statistical significance(P<0.01);the serum TC content in the high frequency group is lower than that in the low frequency group,which has significant statistical significance(P<0.01).(3)Changes in serum LDL-C content:The LDL-C content of the rabbits in the model group were significantly higher than that in the normal group,and there was a significant statistical difference with the rabbits in the normal group(P<0.01);The contents of LDL-C were significantly reduced,compared with the model group,it was statistically significant(P<0.01);the serum LDL-C content in the high frequency group was lower than that in the low frequency group,which was no statistically significant(P>0.05).(4)Changes of serum HDL-C content:The HDL-C content of rabbits in the model group was significantly higher than that in the normal group,and there was a significant statistical difference compared with the rabbits in the normal group(P<0.01);HDL-C content was higher than the model group,which was statistically significant(P<0.05).(5)Serum TC/HDL-C ratio:The model group’s TC/HDL-C ratio was significantly higher than that of the normal group,and there was a significant statistical difference with the normal group rabbits(P<0.01);The serum TC/HDL-C ratio of rabbits was lower than that of the model group,which was statistically significant(P<0.01).(6)Serum LDL-C/HDL-C ratio:The ratio of LDL-C/HDL-C in the rabbits of the model group was significantly higher than that of the normal group.Compared with the rabbits of the normal group,there was a significant statistical difference(P<0.01);high The serum LDL-C/HDL-C ratio of rabbits in the frequency group was lower than that in the model group,which was statistically significant(P<0.01).3.Pathological and ultrastructural changes of rabbit aorta in each groupCompared with the model group,mild moxibustion can alleviate atherosclerotic lipid deposition-like pathological changes in AS rabbits,inhibit vascular intimal damage,and have a certain repair effect on the vascular wall.The ultrastructure of the aorta indicates that mild moxibustion can maintain the integrity and function of endothelial cells.Generally,high-frequency mild moxibustion has a better vascular protection effect than low-frequency subgroups.4.Expression of AMPKa mRNA and protein in rabbit aorta and liver of each groupCompared with the normal group,in the aorta of rabbits of the model group’s expression levels of AMPKa mRNA and protein were reduced,and the differences were statistically significant(P<0.01).The expression levels of AMPKa mRNA and protein in the aorta of rabbits in the moxibustion group were increased.The group ratio and the difference were statistically significant(P<0.01),while the expression of AMPKa mRNA and protein in rabbit aorta in the high frequency group was higher than that in the low frequency group,and the difference was statistically significant(P<0.05,P<0.01).The expression level of liver AMPKa mRNA in the model group rabbits decreased,which was significantly different from that of the normal group(P<0.01),and the expression of liver AMPKa protein was also down-regulated,but there was no significant difference compared with the normal group(P>0.05).The liver AMPKa mRNA and protein expressions of rabbits in the moxibustion group were significantly increased,and there were significant differences compared with the model group(P<0.01).The liver AMPKa mRNA expression was higher in the high frequency group than in the low frequency group,and the difference was statistically significant(P<0.05),but in terms of AMPKa protein expression,there was no statistical significance between the two(P>0.05).5.LXRa mRNA and protein expression levels in rabbit aorta and liver of each groupThe expressions of LXRa mRNA and protein in rabbit aorta in the model group were significantly reduced,and the difference was statistically significant compared with the normal group(P<0.01).The expressions of LXRa mRNA and protein in rabbit aorta in the moxibustion group were significantly up-regulated,which was significantly different from that in the model group(P<0.01).The expression of LXRa mRNA in rabbit aorta in the high frequency group was higher than that in the low frequency group.Significantly statistically significant(P<0.01),but there was no statistical significance between the two in terms of LXRa protein expression(P>0.05).The expression of LXRa mRNA and protein in the liver of the model group decreased significantly,and the differences were significantly statistically significant compared with the normal group(P<0.01).After moxibustion,liver LXRa mRNA and protein expressions increased,and the differences were significantly statistically significant compared with the model group(P<0.01),while rabbit liver LXRa mRNA expression was higher in the high frequency group than in the low frequency group.Compared with the low frequency subgroup,the difference was statistically significant(P<0.01),but in terms of LXRa protein expression,there was no statistical significance between the two(P>0.05).6.ABCA1 mRNA and protein expression levels in rabbit aorta and liver of each groupABCA1 mRNA and protein expressions of rabbit aorta in the model group were significantly reduced,and the difference was significantly statistically significant compared with the normal group(P<0.01).After moxibustion,the expression of ABCA1 mRNA and protein in rabbit aorta increased,which was significantly different from the model group(P<0.01),while the expression of ABCA1 mRNA and protein in the high frequency group was higher than that in the low frequency group.Group,the differences were statistically significant(P<0.01).The expression of ABCA1 mRNA and protein in rabbit liver of the model group was significantly reduced,and there was a significant difference compared with the normal group(P<0.01).The expression of ABCA1 mRNA and protein in rabbit liver was increased after moxibustion,which was significantly different from the model group(P<0.01),while the expression of ABCA1 mRNA and protein in the high frequency group was higher than that in the low frequency group.The differences were statistically significant(P<0.01).Conclusion:1.Mild moxibustion at different frequencies can regulate the blood lipid level of AS rabbits to a certain extent and inhibit aortic endometrial damage.High-frequency mild moxibustion is better than low-frequency mild moxibustion.2.Gentle moxibustion at different frequencies can up-regulate the expression of AMPKa,LXRa and ABCA1 genes and proteins in AS rabbits.And high-frequency mild moxibustion is better than low-frequency mild moxibustion,which may be one of the mechanisms for moxibustion therapy to prevent atherosclerosis. |
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