Study On The Effect Of Acupuncture On Atherosclerosis And Intestinal Microflora In Mice Based On "Heart And Small Intestine"

Objective:This research aims to investigate acupuncture effects at Shu and Mu points on atherosclerosis（AS）and intestinal microflora in mice based on heart and small intestine,and to provide experimental basis for acupuncture prevention of AS and regulation of intestinal microecology.Methods:Nineteen healthy 8-week-old male C57BL/6J mice were selected as normal control group（NC group）and give basic feeding.64 apolipoprotein E gene knockout(Apo E^-/-)mice with the same genetic background were given high fat feeding as model group（MM group）.After 12 weeks of continuous feeding,4 mice from each group were taken for HE staining of aortic root to identify the results of modeling.After successful modeling,60 mice in MM group were randomly divided into 4 groups:model group（M group）,acupuncture group（AC group）,positive drug group（PD group）,acupuncture combined with positive drugs group（ACPD group）,15 mice in each group.No intervention was conducted in NC and M.AC group was treated with acupuncture at small intestine Shu Mu points once a day,6 days as a course of treatment,1 day off between courses,a total of 4 courses of treatment;PD was treated with atorvastatin calcium solution by gavage once a day,and the course of treatment was the same as that in AC;ACPD was treated with acupuncture at small intestine Shu Mu points and atorvastatin calcium solution by gavage,and the course of treatment was the same as that in AC.After 4 courses of treatment,serum TG,TC,LDL-C and HDL-C of the mice were detected to observe the difference of blood lipid levels in each group;HE and oil red O staining were used to observe the pathological morphological changes and the difference of fat content in the aortic root of mice;DNA was extracted from feces for 16Sr DNA detection to observe the changes of intestinal microflora structure and abundance in each group.Results:After 12 weeks of high fat feeding,HE staining of aortic root of mice in MM showed thickening of inner wall,more foam cells and stenosis of lumen,and successful replication of AS model.After 4 weeks’intervention,（1）Blood lipid test results:Compared with NC gorup,TG,TC and LDL-C of mice in M group were increased,while HDL-C level was decreased,the difference was statistically significant（P<0.05）;Compared with M group,TG,TC and LDL-C in PD and ACPD groups were decreased,while HDL-C level was increased,the difference was statistically significant（P<0.05）.However,there was no statistical significance in TG,TC,LDL-C and HDL-C levels between AC group and M group（P>0.05）.（2）HE staining:Compared with NC group,the intima of blood vessel of mice in M group were severely thickened,with a large number of foam cells aggregated in the inner wall with calcification,and severe stenosis of vascular lumen;Compared with M group,the intima thickness of mice in AC,PD and ACPD groups were reduced,and the foam cells were reduced.Among them,the mice in ACPD group showed the most obvious reduction in intima thickness and the least number of foam cells.（3）Oil red O staining:Compared with NC group,the intima of blood vessels of mice in M group were severely thickened,with a large amount of lipid deposition and foam cell aggregation;Compared with M group,the lipid deposition and foam cells of mice in AC,PD and ACPD groups were significantly reduced.Among them,the mice in ACPD group had the least lipid deposition and the least number of foam cells in the intima of blood vessels.（4）Detection of intestinal microflora:there were significant differences among groups in phylum,family,genus and species levels.At the phylum level,compared with NC group,firmicutes and proteus increased and bacteroidetes decreased in M group（P<0.05）;Compared with M group,bacteroidetes increased,firmicutes and proteusdecreased in AC,PD and ACPD groups（P<0.05）.Among them,bacteroidetes accounted for the highest proportion in ACPD group and the lowest in M group;firmicutes accounted for the highest in group M and the lowest in group ACPD;proteobacteria accounted for the highest proportion in group M and the lowest in group AC.Conclusion:Acupuncture at Shu and Mu points of small intestine can improve blood lipid and reduce atherosclerosis in AS model mice,and regulate the abundance and structure of intestinal flora in AS model mice.The effective way to improve AS is to regulate intestinal flora by acupuncture at Shu and Mu points of small intestine.