Wheat Grain Moxibustion Up Regulates TFEB Through AMPK/mTOR Signaling Pathway To Improve Hepatocyte Autophagy In Hyperlipidemia Rats

Objective:To explore the molecular mechanism of wheat-grain moxibustion to improve hepatocyte autophagy in the case of abnormal blood lipids.Based on the AMPK/mTOR signaling pathway,explore the molecular biological mechanism of grain moxibustion by influencing TFEB and regulating liver cell autophagy in hyperlipidemia SD rats.Methods:In this study,SD rats(n=30)were used as the research object.Four blood lipids(LDL,TC,TG,HDL)were detected by ELISA after 8 weeks of high-fat diet.Experimental groupings:Blank group,HFD group,HFD+Drug group,HFD+CC+Moxi group and grain moxibustion intervention group(HFD+Moxi group)were used for control,with 6 rats in each group.Intervention methods:Blank group was fed basic feed with unlimited water;HFD group was fed high-fat feed with unlimited water;HFD+Drug group was fed high-fat feed with unlimited water,atorvastatin gavage;HFD+Moxi The group was fed high-fat feed with unlimited water,and moxibustion with grains stimulated the bilateral Hou Sanli(3 Zhuang/acupoint);the HFD+CC+Moxi group was fed high-fat feed with unlimited water,and the compound C solution was injected intraperitoneally."Housanli" wheat grain moxibustion stimulation(3 Zhuang/acupoint).In terms of the intervention frequency and cycle,the frequency of the intervention measures remained the same,once per day,with two days off for every 5 days of intervention,for a total of 10 weeks of intervention.Test items:1.Enzyme-linked immunosorbent assay to detect serum HDL,LDL,TC,and TG levels in SD rats;2.Hepatocyte oil red O staining to assess liver lipid deposition;3.Immunohistochemical method to detect liver tissue TFEB Expression distribution;4.Immunofluorescence method to detect the expression distribution of LC3,p62,TFEB in the liver tissue of each group;5.Western blotting method to detect the expression of liver tissue LC3Ⅰ,LC3Ⅱ,p62,AMPK,mTOR,TFEB;6.Real-time quantitative aggregation The expression of AMPK,mTOR,TFEB mRNA in rat liver tissues was detected by ELISA method.Results:1.Bilateral "Housanli" wheat grain moxibustion can increase the expression of serum HDL and reduce the serum LDL,TC,TG expression in hyperlipidemia SD rats;2."Housanli" wheat grain moxibustion can reduce the liver slices The accumulation of fat in the liver can improve the steatosis of liver cells;3.Mixed moxibustion to stimulate "Hou Sanli" can increase the number and protein expression of LC3 positive cells in the liver of hyperlipidemia SD rats,and reduce the number and protein expression of P62 positive cells in the liver,and significantly increase The level of autophagy in the liver;4.Mixed moxibustion "Hou Sanli"stimulation can increase the protein and mRNA expression of AMPK,inhibit the protein and mRNA expression of mTOR,and increase the number of positive nuclei,positive cell count,protein and mRNA expression of TFEB.Conclusion:"Housanli" grain moxibustion can improve the blood lipid level of hyperlipidemia SD rats,increase the expression of TFEB by activating the liver AMPK/mTOR signaling pathway,and initiate the transcription of autophagy genes such as LC3 and lysosomes Synthesize to increase the autophagy level of rat liver cells,thereby improving liver cell damage and lipid metabolism.