The Effect And Mechanism Of Small Molecule Compound Z526 In Treating Cancer Cachexia

Background: Cancer cachexia is a systemic wasting syndrome characterized by weight loss,skeletal muscle atrophy and fat loss,which cannot be completely reversed by routine nutritional support.The occurrence of cancer cachexia severely worsens the quality of living and treatment compliance of cancer patients,leading to about 20% of cancer patients’ deaths.The pathogenesis of cancer cachexia is very complex and has not yet been fully understood.Currently,there is still no effective drug on the market to treat cancer cachexia in clinical practice.Pyrrolidine dithiocarbamate(PDTC)can effectively alleviate cancer cachexia.Small molecule compound Z526 is a PDTC modification obtained by structural modification to solve the problems of oral inefficacy and toxicity of PDTC.However,the effect and mechanism of Z526 in the treatment of tumor cachexia are still unclear,and further research is needed.Purpose: In this study,the effect and mechanism of Z526 in alleviating cancer cachexia were investigated through in vivo and in vitro experiments,and the effect of Z526 on cancer cachexia treated with chemotherapeutic drugs was evaluated.Methods:(1)C2C12 muscle atrophy and 3T3-L1 adipocyte degradation model of cancer cachexia in vitro were established by induction of C26 cell medium,TNF-αand IL-6;(2)MTT assay was used to detect cell viability;(3)Cell H&E staining was performed,and the diameter of C2C12 myotube diameter was calculated by Image J;(4)Oil red O staining was used to evaluate lipid droplet size;(5)The contents of glycerol and triglyceride in 3T3-L1 adipocytes were detected by glycerol and triglyceride detection kits;(6)In vivo C26 cancer cachexia mouse model was used to evaluate the effect of Z526 on cancer cachexia and the effect of Z526 combined with oxaliplatin(OX)on tumor cachexia;(7)H&E staining of tissue was performed,and the muscle bundle and fat area were calculated by Image J;(8)The contents of glycerol and triglyceride in the serum of mice were detected by glycerol and triglyceride detection kits;(9)The levels of TNF-α and IL-6 in the serum of mice were detected by ELASA kit;(10)Western Blot was used to detect protein levels in cells and tissues.Results:(1)Z526 can effectively alleviate the atrophy of C2C12 myotubes induced by C26 medium,and the diameter of C2C12 myotubes gradually added with the add of drug concentration with a concentration gradient;The mechanism may be that Z526 promotes the synthesis of muscle protein by promoting the phosphorylation of Akt and increasing the expression of MHC and Myo D.Meanwhile,NF-κB and MAPK signaling pathways were inhibited to down-regulate the expression of E3 ubiquitin ligase MAFBX and inhibit the degradation of muscle protein.(2)Z526 can effectively alleviate the 3T3-L1 adipocyte degradation induced by C26 medium,and inhibit the decomposition of triglycerides and the release of free glycerol in a concentration-dependent manner.The mechanism may be that Z526 inhibit NF-κB,MAPK and AMPK signaling pathways,inhibit the activation of HSL and inhibit fat mobilization.At the same time,the expression of UCP1 was down-regulated to reduce ineffective thermogenesis and inhibit fat browning.(3)Z526 can relieve cancer cachexia symptoms in C26 tumor-bearing mice.At the end of the trial,Z526 alleviated weight loss,muscle atrophy and adipose tissue degradation without affecting tumor size,and was more effective in adipose tissue.The mechanism of Z526 in muscle and adipose tissue of mice was studied,and the results were similar to those in vitro.(4)Z526 significantly inhibited the increase of serum inflammatory cytokines TNF-α and IL-6 in C26 tumor-bearing mice.(5)Z526 effectively alleviated the atrophy of C2C12 myotubes induced by TNF-α and IL-6 in a concentration-dependent manner.The mechanism of Z526 alleviates muscle atrophy may be that TNF-α regulates NF-κB and MAPK signaling pathways,while IL-6regulates STAT-3 and AKT signaling pathways.(6)Z526 effectively alleviated3T3-L1 adipocyte degradation induced by TNF-α and IL-6 in a concentration-dependent manner.The mechanism of Z526 alleviating fat degradation may be that TNF-α regulates NF-κB and MAPK signaling pathways,while IL-6regulates AMPK signaling pathway.(7)Z526 can relieve the tumor cachexia symptoms aggravated by OX in C26 tumor-bearing mice without affecting the therapeutic effect of OX.Conclusion: Z526 can alleviate muscle atrophy and fat degradation of cancer cachexia by affecting inflammatory factors TNF-α and IL-6.Z526 is expected to be a potential drug for clinical treatment of tumor cachexia.The systematic preclinical development of Z526 is conducive to the research and development of new anti-cancer cachexia drugs with independent intellectual property rights,and promotes the application of basic research results.