peoniflorin and Diabetes-Mellitus--Type-2

Investigation of the synergistic and complementary effects is vital but difficult for Chinese herbal medicine. We explored the synergistic and complementary mechanisms of berberine (BBR) and paeoniflorin (PF) in the treatment of type 2 diabetes mellitus (T2DM) through network pharmacology and molecular docking. We identified putative targets of BBR, PF, and T2DM, and constructed a protein-protein interaction (PPI) network. Gene ontology and Kyoto encyclopedia of gene and genomes pathway enrichment analysis and molecular docking were used to predict the molecular mechanisms. A diabetes model was induced by a high-fat diet to verify the therapeutic effect. Ninety-two targets of BBR + PF in the treatment of T2DM were identified, which were considered as synergistic targets. Fifty-nine complementary targets of BBR-T2DM and 47 of PF-T2DM were identified. PPI network analysis showed that JAK2, ESR1, IFG1R, STAT3, EGFR, MAPK1, and AKT1 are closely related to T2DM. The enrichment analysis further showed that the synergistic targets mainly involved the AGE-RAGE signaling pathway in diabetic complications, FOXO, AMPK, and VEGF signaling pathways, and glycolysis/gluconeogenesis. AKT1, JAK2, and STAT3, which are common targets of the AGE-RAGE signaling pathway in diabetic complications and the FOXO signaling pathway, were chosen for docking with BBR and PF, respectively, and showed good binding activities. BBR + PF significantly reduced weight and fasting blood glucose, and alleviated insulin resistance. Moreover, BBR + PF promoted the phosphorylation of AKT1, JAK2, and STAT3. This study provides information to understand the synergistic and complementary mechanism of BBR + PF against T2DM, and may facilitate the development of new anti-T2DM drugs.

Topics: Animals; Berberine; Blood Glucose; Diabetes Mellitus, Type 2; Disease Models, Animal; Drugs, Chinese Herbal; Glucosides; Humans; Hypoglycemic Agents; Male; Mice; Mice, Inbred C57BL; Molecular Docking Simulation; Monoterpenes; Network Pharmacology; Phytotherapy

Diabetic liver injury (DLI) is a complication that damages the quality of life in diabetes patients. While paeoniflorin (PF) exhibits anti-inflammatory and antioxidant effects, no data are available on whether PF protects against DLI. Therefore, we evaluated the effects of PF on hepatic steatosis and inflammation in db/db mice, a type 2 diabetes model.. In this study, we investigated the effects of PF on DLI using diabetic mice model (db/db mice) and high glucose (HG)-induced mouse AML12 cells. The effects of PF on TXNIP-mediated NLRP3 inflammasome in vivo and in vitro were evaluated by Western bloting, RT-PCR, immunohistochemistry (IHC) and immunofluorescence (IF) analysis. Through molecular docking experiments and cellular thermal shift assay (CETSA), we studied the binding ability of PF to thioredoxin-interacting protein (TXNIP). We use TXNIP siRNA to knock down TXNIP in AML12 cells.. We found that PF reversed abnormal liver function and liver steatosis in db/db mice, while blocking the release of inflammatory cytokines. These effects are associated with PF inhibition of the TXNIP/NLRP3 signaling pathway. Molecular docking experiments and CETSA also demonstrated that TXNIP is a likely target of PF. In HG-treated AML12 cells, TXNIP knockdown eliminated the beneficial effects of PF.. Using a combination of animal and in vitro experiments, this study demonstrated for the first time that PF ameliorates DLI through targeting the TXNIP-activated NLRP3 inflammasome. Thus, PF may be a potential therapeutic agent against DLI.

Topics: Animals; Carrier Proteins; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Fatty Liver; Glucosides; Inflammasomes; Mice; Molecular Docking Simulation; Monoterpenes; NLR Family, Pyrin Domain-Containing 3 Protein; Reactive Oxygen Species; Thioredoxins

Paeoniflorin is an effective Chinese traditional medicine with anti-inflammatory and immune-regulatory effects. The aim of this study was to investigate the underlying renoprotective mechanism of Paeoniflorin. In vivo, db/db mice were intraperitoneally injected with Paeoniflorin at a dose of 15, 30, or 60 mg/kg respectively. The immunostaining of TLR2, TLR4, CD68, NF-kB p65 and the mRNA level of inflammatory factors, together with the protein expression of TLR2/4 signaling were evaluated. Our data demonstrated that Paeoniflorin could decrease the urinary albumin excretion rate and inhibit macrophage infiltration and activation through blockage of the TLR2/4 signaling pathway compared with the db/db group in vivo. In vitro, RAW264.7 cells were categorized into control, bovin serum albumin (BSA)-stimulated, advanced glycation end products (AGEs)-stimulated, Paeoniflorin intervention and oxidized phospholipid (OxPAPC)-inhibited groups. The cell viability, the optimal stimulated time and concentration were measured as well as the TLR2/4 signaling activation determined by RT-PCR, Western blot and ELISA. Our data demonstrated that Paeoniflorin reduced the AGEs-induced TLR2/4 activation and inflammatory responses, which was consistent with the TLR2/4 inhibitor group. These findings indicate that Paeoniflorin prevents macrophage activation via inhibition of TLR2/4 signaling expression in type 2 diabetic nephropathy.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Cells, Cultured; China; Diabetes Mellitus, Type 2; Diabetic Nephropathies; Glucosides; Medicine, Chinese Traditional; Mice; Monoterpenes; Protective Agents; Signal Transduction; Toll-Like Receptor 2; Toll-Like Receptor 4