The Mechanism Of Baoyuan Jiedu Decoction On Liver Lipid Metabolism And Liver Mitochondrial Function In Carcinomatous Cachexia Mice

Objective:This study aims to investigate the effect and mechanism of Baoyuan Jiedu Decoction on improving mitochondrial energy metabolism and lipid metabolism in the liver of C57/BL LLC homologous xenograft tumor cachexia mice using lipidomics,molecular biology,and other research methods.The study provides experimental evidence for the effective use of traditional Chinese medicine to correct metabolic disorders in the liver of cancer cachexia.Methods:After 4 weeks of intervention with Baoyuan Jiedu Decoction in C57 tumorbearing mice,the general condition and mental status of the animals,food intake,body weight,tumor weight,tumor mass,and weight loss rate were observed.Liver tissue was stained with HE and oil red O,and the organ index of liver and spleen was calculated to evaluate the intervention effect of Baoyuan Jiedu Decoction on multiple organs and quality of life in the CC model mice.Multi-dimensional mass spectrometry shotgun lipidomics(MDMS-SL)technology was used to analyze changes in 11 types of lipid molecules,including CL,TAG,PI,PC,PE,LPC,SM,LPC,Cer,PA,and FA,in the liver tissue of CC model mice.Metabo Analyst 5.0 metabolomics analysis tool was used to perform PCA analysis of the total lipid class and statistical analysis of various lipid molecules,to investigate the mechanism of Baoyuan Jiedu Decoction in improving liver lipid metabolism disorders.Using liver mitochondrial function as the entry point,electron microscopy was used to observe liver mitochondrial morphology,and the ATP content in mouse liver tissue was detected by colorimetric assay.Western blot was used to detect changes in the protein content of liver mitochondrial respiratory chain complexes Ⅰ-Ⅳ and mitochondrial lipid ceramide transferase ALCAT1,to evaluate the protective effect of Baoyuan Jiedu Decoction on liver mitochondrial function and predict potential novel mitochondrial lipid biomarkers for cancer cachexia.Result:The mouse model of cachexia was successfully established,and after intervention with Baoyuan Jiedu Decoction,compared with the model group,the Baoyuan Jiedu Decoction group and megestrol acetate group showed decreased body weight,tumor weight and weight loss rate(P<0.05),increased food intake(P<0.05),reduced tumor weight(P<0.05),relieved liver atrophy and spleen enlargement(P<0.05),reduced liver inflammation pathology and hepatic steatosis(P<0.05),and significant improvement in overall survival quality and mental status.Baoyuan Jiedu Decoction significantly improved the disorder of lipid metabolism in tumor-bearing mice.The lipid content of each group was analyzed by MDMS-SL shotgun multidimensional mass spectrometry,and principal component analysis of differentially expressed lipids before and after intervention was performed using MetaboAnalyst 5.0 online tool,which found that 18 lipid molecules and their metabolites were associated with the progression of cachexia,including CL cardiolipin,SM sphingomyelin,CER ceramide,FA fatty acid,TAG triglyceride,LPC lysophosphatidylcholine,PC phosphatidylcholine,etc.Several lipid subcategories confirmed the "lipid synthesis phenotype" unique to cachexia development.Among them,the content of mitochondria-specific lipid cardiolipin(Cardiolipin)increased and the composition of unsaturated fatty acid chains increased.CL(tri-18:2,tri-16:1,tri-18:1)was predicted as a potential lipid biomarker for BJD against CC.Baoyuan Jiedu Decoction significantly increased ATP content in mouse liver(P<0.05);the protein expression levels of liver mitochondrial respiratory chain complexes Ⅰ-Ⅳ:SDHB/UQCRC2/MT-CO1/NDFUB8 were significantly increased,indicating the improvement of liver mitochondrial oxidative phosphorylation function in CC mice.Transmission electron microscopy showed that although some mitochondrial contents were still degraded after drug intervention,mitochondrial vacuolization,mitochondrial morphology distortion,and ridge damage distortion were reduced,indicating that autophagosomes increased in liver cells,suggesting that the drug promotes the clearance of damaged mitochondria to achieve mitochondrial quality control.Conclusion:Baoyuan Jiedu Decoction can significantly improve the survival quality and general condition of cachectic mice and exert a protective effect on improving liver inflammation and preventing lipid droplet loss.Based on MDMS-SL technology analysis of the CC model mouse liver lipid metabolism,including TAG,FA,PC,SM,PS,PE,LPC,Cer,CL,PA,PI;a total of 11 lipids,the mechanism of Baoyuan Jiedu Decoction in treating CC mice may be through improving phospholipid metabolism in the body,and phospholipids may be a potential new biomarker of cachexia.Baoyuan Jiedu Decoction promotes the increase of mitochondrial ATP content in cancer cachexia mice liver,clears damaged mitochondria,and increases liver oxidative phosphorylation level.The content of liver phospholipids is positively correlated with liver mitochondrial oxidative phosphorylation before and after traditional Chinese medicine intervention,which proves that Baoyuan Jiedu Decoction intervenes in the synthesis of liver phospholipids to improve liver mitochondrial function in the cancer cachexia model.