Study Of The Effects Of Moxibustion With Seed-Sized Moxa Cone At Fenglong On Hyperlipidemia By Regulating Lipid Metabolism Based On Plasma Metabolomics

OBJECTIVE:This study intends to analyze the potential metabolites and metabolic pathways involved in hyperlipidemia treatment by Moxibustion with seed-sized moxa cone using metabolomics method based on objectively evaluating the clinical efficacy of moxibustion at Fenglong for treating hyperlipidemia,in order to explore the effect mechanism of moxibustion with seed-sized moxa cone for for treating hyperlipidemia,so as to provide a reliable scientific basis for the clinical extension application of with seed-sized moxa cone treating hyperlipidemia.It also provides a new research direction and research idea for further research on the action mechanism of moxibustion method.Methods:A total of 114 hyperlipidemia patients were randomly divided into three groups,hyperlipidemia control group(blank group for short),drug control group(drug group for short,oral Xuezhikang Capsule)and moxibustion with seed-sized moxa cone treatment group(treatment group for short),38 patients in each group.2 cases were dislodged in the blank group,36 cases actually;One case was dislodged in the drug group,37 cases actually;2 cases were dislodged in the treatment group,cases actually during the study period.Patients included in the treatment group were treated by moxibustion with seed-sized moxa cone twice a week for 8 consecutive weeks at Fenglong.Patients in the drug group received 8-week continuous oral administration of Xuezhikang Capsules,2 pills twice daily.Patients in the blank group did not do any treatment during this period.Evaluation time points including baseline period and after 8 weeks of intervention treatment,the levels of total cholesterol(TC),triglycerides(TG),low-density lipoprotein cholesterol(LDL-C)and high-density lipoprotein cholesterol(HDL-C)in three groups were detected.Another group of healthy people(HC)of the same age were recruited for comparison with hyperlipidemic control patients(HLP),and patients in the treatment group were compared before and after treatment(BMT before moxibustion treatment,AMT after moxibustion treatment),differential metabolites between groups were identified using liquid chromatography-mass spectrometry(LC-MS)tandem analysis,and common differential metabolites between the two comparison groups,HLP vs HC and AMT vs BMT,were analyzed for their enriched pathways.Twenty hyperlipidemic mice were constructed and randomly divided into 10 in model group and 10 in treatment group,compared with 10 normal mice.Mice in the treatment group were subjected to moxibustion with seed-sized moxa cone at Fenglong intervention every other day for 2 weeks.Liver histomorphological changes in mice were observed by HE staining and oil red O staining,and the expression of microtubule associated protein LC3B(a marker of autophagy)in hepatocytes was observed by immunohistochemistry,to initially explore whether moxibustion with seed-sized moxa cone exerts effect on autophagy regulation and then improves hyperlipidemia.Result:1.Overall response rate:3 patients in the treatment group were clinically controlled,12 showed efficacy,15 were effective and 6 were ineffective,the overall response rate was 83.33%.One patient in the drug group was clinically controlled,10 showed efficacy,21 were effective and 5 were ineffective,with an overall response rate of 86.49%.The difference between the two groups was not statistically significant(P>0.05).2.Lipid improvement:Compared with the baseline period,there were no significant differences in TC(P>0.05),TG(P>0.05),HDL-C(P>0.05),and LDL-C(P>0.05)in blank group patients after 8 weeks;Compared with the baseline period,patients in the treatment group had lower TC(P<0.01),TG(P<0.01),LDL-C(P<0.01)and higher HDL-C(P<0.01),with statistically significant differences after 8 weeks;Compared with the baseline period,patients in the drug group had lower TC(P<0.01),TG(P<0.01),LDL-C(P<0.07)with statistically significant differences,with no significant difference in HDL-C(P>0.05)after 8 weeks.Compared with the blank group,patients in treatment group had lower TC(P<0.01),TG(P<0.01),LDL-C(P<0.01)and higher HDL-C(P<0.05),with statistically significant differences;Compared with the blank group,patients in drug group had decreased TC(P<0.01),TG(P<0.01),LDL-C(P<0.01),with statistically significant differences,HDL-C(P>0.05)levels were not significantly different;Compared with the drug group,the levels of TC(P>0.05),TG(P>0.05),LDL-C(P>0.05),HDL-C(P>0.05)did not differ significantly between the treatment group.3.Metabolomic analysis:87 and 51 differential metabolites were identified in HLP vs HC and AMT vs BMT.There were 21 differential metabolites with opposite altered relationships between the two compared groups.The differential metabolites in HLP vs HC group were significantly enriched in glycerophospholipid metabolism,sphingolipid metabolism and other metabolic pathways.The differential metabolites in AMT vs BMT group were significantly enriched in glycerophospholipid metabolism,fatty acid biosynthesis,unsaturated fatty acid biosynthesis,autophagy regulation,sphingolipid metabolism,linoleic acid metabolism and other metabolic pathways.Among them,glycerophospholipid and sphingolipid metabolic pathways were simultaneously present between the two compared groups.4.Animal experiment showed that:A large amount of lipid deposition was found in hepatocytes of hyperlipidemia mice,which was markedly reduced after moxibustion with seed-sized moxa cone intervention;The expression of lc3b in hepatocytes of hyperlipidemia mice was decreased compared with normal group,which was increased after moxibustion with seed-sized moxa cone intervention.Conclusion:1.Moxibustion with seed-sized moxa cone is an effective therapy for treating hyperlipidemia.After treatment,the levels of TC,TG,LDL-C decreased and HDL-C increased in patients,and this lipid-lowering effect was comparable with the efficacy of oral Xuezhikang.2.Moxibustion with seed-sized moxa cone affected lipid metabolism related pathways,especially glycerophospholipid metabolism and sphingolipid metabolism,to repair lipid metabolic network disturbance and regress metabolic profiles toward the normal range,resulting in a therapeutic effect on hyperlipidemia;3.Moxibustion with seed-sized moxa cone for hyperlipidemia may be associated with initiating regulation of autophagy,which can improve hyperlipidemia by activating autophagy,promoting hepatic lipid metabolism and reducing hepatic lipid deposition;4.Moxibustion with seed-sized moxa cone treating hyperlipidemia deserves to be promoted and applied in clinic.