demethyleneberberine and Non-alcoholic-Fatty-Liver-Disease

Demethylenetetrahydroberberine (DMTHB) is a novel derivative of berberine and demethyleneberberine. This research explored the pharmacological effects and molecular mechanisms of DMTHB on nonalcoholic fatty liver disease (NAFLD).. C57BL/6 mice were induced by a methionine- and choline- deficient (MCD) diet and L02 cells were induced by palmitic acid to establish NAFLD animal and cell models. qPCR and western blotting were used to detect the expression of genes and proteins associated with pharmacological mechanism. A biotin-labeled DMTHB pulldown assay was used to further clarify the pharmacological targets.. Our results indicated that DMTHB significantly alleviates NAFLD in mice. Biochemical assays showed that serum alanine aminotransferase, aspartate aminotransferase and hepatic lipids were significantly decreased in MCD-induced NAFLD mice orally administered of DMTHB (50 mg/kg or 150 mg/kg body weight daily) for 30 d. qPCR and ELISA analysis demonstrated that DMTHB reduced the expression of serum proinflammatory cytokines, such as TNF-α, IL-1β and IL-6. Moreover, pull-down assays and compound-centric chemical proteomics illustrated that DMTHB inhibited NOD-like receptor protein 3 (NLRP3) inflammasome signaling. In addition, DMTHB also attenuated oxidative stress and endoplasmic reticulum stress by downregulation CYP2E-1 and ATF-4 expression. Moreover, DMTHB treatment ameliorated the liver fibrosis in MCD-induced NAFLD mice by suppressing the expression of TGF-β1, α-SMA and collagen 1A1.. DMTHB targeted the NLRP3 inflammasome to suppress inflammation and inhibited CYP2E1 to reduce oxidative stress and ER stress. Consequently, DMTHB may have therapeutic benefits in the treatment of NAFLD in the clinic.

Topics: Animals; Berberine; Humans; Inflammasomes; Male; Mice; Mice, Inbred C57BL; NLR Family, Pyrin Domain-Containing 3 Protein; Non-alcoholic Fatty Liver Disease; Oxidative Stress

Non-alcoholic fatty liver disease (NAFLD) has reached an epidemic level globally, which is recognized to form non-alcoholic steatohepatitis (NASH) by the "two-hit" model, including oxidative stress and inflammation. AMP-activated protein kinase (AMPK) has long been regarded as a key regulator of energy metabolism, which is recognized as a critical target for NAFLD treatment. Here we introduce a natural product, demethyleneberberine (DMB), which potentially ameliorated NAFLD by activating AMPK pathways. Our study showed that the intraperitoneal injection of DMB (20 or 40 mg/kg body weight) decreased hepatic lipid accumulation in methionine and choline deficient (MCD) high-fat diet feeding mice and db/db mice. The further investigation demonstrated that DMB activated AMPK by increasing its phosphorylation in vitro and in vivo. Accompanied with AMPK activation, the expression of lipogenic genes were significantly reduced while genes responsible for the fatty acid β-oxidation were restored in DMB-treated NAFLD mice. In addition, the remarkable oxidative damage and inflammation induced by NAFLD were both attenuated by DMB treatment, which is reflected by decreased lipid oxidative product, malonaldehyde (MDA) and inflammatory factors, tumor necrosis factor α (TNFα) and interleukin 1β (IL-1β). Based on all above, DMB could serve as a novel AMPK activator for treating NAFLD and preventing the pathologic progression from NAFLD to NASH by inhibiting the oxidative stress and inflammation.

Topics: AMP-Activated Protein Kinases; Animals; Antioxidants; Berberine; Enzyme Activation; Hep G2 Cells; Humans; Lipid Metabolism; Liver; Male; Mice, Inbred C57BL; Mice, Inbred ICR; Non-alcoholic Fatty Liver Disease; Oxidative Stress