Grey Relational Analysis Combined With Network Pharmacology to Identify Antioxidant Components and Uncover Its Mechanism From Moutan Cortex

The present study determines the potential antioxidants in Moutan Cortex (MC) and predicts its targets of anti-oxidative activities. The quantitative analysis and the free radical scavenging assays were conducted to detect the main components in MC and assess its anti-oxidant activities. The grey relational analysis and the network pharmacology approach were employed to predict its key components and targets of anti-oxidant activities. Six main constitutes in MCs were quantified by high performance liquid chromatography (HPLC) and its anti-oxidant activities were evaluated by DPPH and ABTS free radical scavenging methods. Then grey relational analysis was employed to predict the key components acting on anti-oxidative activity based on the chem-bio results. The predicted components and its mechanisms on anti-oxidation were uncovered by network pharmacology approach and cell test, respectively. The content of paeonol and paeoniflorin accounts for more than 80% the whole content of detected components. However, the two main ingredients showed a great variety among MCs. The antioxidant capacities of MCs also showed a great discrepancy based on DPPH and ABTS methods. The key components acting on anti-oxidation were identified to be paeonol, gallic acid and benzoylpaeoniflorin, and their potential therapeutic targets were predicted and verified, respectively. The present results reveal that MC has a significant antioxidant activity and the compounds of paeonol, gallic acid and benzoylpaeoniflorin could be considered as the promising antioxidant candidates with the property of suppressing oxidative stress and apoptosis.

Dietary Moutan Cortex Radicis Improves Serum Antioxidant Capacity and Intestinal Immunity and Alters Colonic Microbiota in Weaned Piglets

Background:Moutan cortex radicis (MCR), as a common traditional Chinese medicine, has been widely used as an antipyretic, antiseptic, and anti-inflammatory agent in China.Objectives: This study aimed to investigate the effects of dietary MCR supplementation on the antioxidant capacity and intestinal health of the pigs and to explore whether MCR exerts positive effects on intestinal health via regulating nuclear factor kappa-B (NF-κB) signaling pathway and intestinal microbiota.Methods: MCR powder was identified by LC-MS analysis. Selected 32 weaned piglets (21 d of age, 6.37 ± 0.10 kg average BW) were assigned (8 pens/diet, 1 pig/pen) to 4 groups and fed with a corn-soybean basal diet supplemented with 0, 2,000, 4,000, and 8,000 mg/kg MCR for 21 d. After the piglets were sacrificed, antioxidant indices, histomorphology examination, and inflammatory signaling pathway expression were assessed. The 16s RNA sequencing was used to analyze the effects of MCR on the intestinal microbiota structure of piglets.Results: Supplemental 4,000 mg/kg MCR significantly increased (P < 0.05) the average daily weight gain (ADG), average daily feed intake (ADFI), total antioxidative capability, colonic short-chain fatty acids (SCFA) concentrations, and the crypt depth in the jejunum but decreased (P < 0.05) the mRNA expression levels of interferon γ, tumor necrosis factor-α, interleukin-1β, inhibiting kappa-B kinase β (IKKβ), inhibiting nuclear factor kappa-B (IκBα), and NF-κB in the jejunum and ileum. Microbiota sequencing identified that MCR supplementation significantly increased the microbial richness indices (Chao1, ACE, and observed species, P < 0.05) and the relative abundances of Firmicutes and Lactobacillus (P < 0.05), decreased the relative abundances of Bacteroides, Parabacteroides, unidentified_Lachnospiraceae, and Enterococcus (P < 0.05) and had no significant effects on the diversity indices (Shannon and Simpson, P > 0.05). Microbial metabolic phenotypes analysis also showed that the richness of aerobic bacteria and facultative anaerobic bacteria, oxidative stress tolerance, and biofilm forming were significantly increased (P < 0.05), and the richness of anaerobic bacteria and pathogenic potential of gut microbiota were reduced (P < 0.05) by MCR treatment. Regression analysis showed that the optimal MCR supplemental level for growth performance, serum antioxidant capacity, and intestinal health of weaned piglets was 3,420 ~ 4,237 mg/kg.Conclusions: MCR supplementation improved growth performance and serum antioxidant capacity, and alleviated intestinal inflammation by inhibiting the IKKβ/IκBα/NF-κB signaling pathway and affecting intestinal microbiota in weaned piglets.

Dietary Moutan Cortex Radicis Alters Serum Antioxidant Capacity, Intestinal Immunity and Colonic Microbiota in Weaned Piglets

Background: Moutan cortex radicis (MCR), as a common traditional Chinese medicine, has been widely used as antipyretic, antiseptic and anti-inflammatory agent in China. However, few studies have evaluated the positive effects of MCR, as a new feed additives, on alleviating weaning stress and improving intestinal health and microbiom in pigs. This study aimed to investigate the effect of dietary MCR supplementation on serum antioxidant capacity, intestinal morphology, anti-inflammatory mechanism, and microbiota in weaned piglets.Results: Supplemental 2000 mg/kg and 4000 mg/kg MCR increased (P < 0.05) the final body weight, ADG and ADFI of weaned piglets, and 2000 mg/kg MCR diet significantly decreased (P < 0.05) the F/G ratio and increased (P < 0.05) serum catalase activity compared with CON group. Also, the villus height and crypt depth in the ileum and the concentrations of total SCFA, acetic acid, butyric acid and valeric acid in the colonic contents were higher (P < 0.05) in the 2000 mg/kg and 4000 mg/kg MCR diets than CON group. Dietary MCR supplementation at 4000 mg/kg MCR significantly increased (P < 0.05) total antioxidative capability and the crypt depth in the jejunum but decreased (P < 0.05) the mRNA expression levels of Interferon γ, tumor necrosis factor-α, interleukin-1β, inhibiting kappa B kinase β (IKKβ), inhibiting nuclear factor kappa-B (IκBα) and nuclear factor kappa-B (NF-κB) in the jejunum and ileum. Supplemental 8000 mg/kg MCR had the higher total antioxidative capability and catalase activity in the serum but decreased (P < 0.05) the villus height and crypt depth in the jejunum compared with the CON group. MCR addition reduced (P < 0.05) serum malondialdehyde content, and tended to increase the mRNA expression of zonula occludens-1 in the ileum (P = 0.066) compared to the CON group. Microbiota sequencing identified the microbial richness indices (Chao1, ACE, and observed species), the relative abundances of Firmicutes and Lactobacillus were increased (P < 0.05), and the relative abundances of Bacteroides, Parabacteroides, unidentified_Lachnospiraceae and Enterococcus were reduced (P < 0.05) by MCR supplemented. Microbial metabolic phenotypes analysis also showed that the richness of aerobic bacteria and facultative anaerobic bacteria, oxidative stress tolerance, and biofilm forming were significantly increased (P < 0.05), and the richness of anaerobic bacteria and pathogenic potential of gut microbiota were reduced (P < 0.05) by MCR treatment. Conclusions: In antibiotic-free diets, MCR supplementation improved growth performance and serum antioxidant capacity, alleviated intestinal inflammatory by inhibiting IKKβ/IκBα/NF-κB signaling pathway and affecting intestinal microbiota in weaned piglets.

Combinatorial Synthesis of A Series of Paeonol-based Phenylsulfonyl hydrazone Derivatives as Insecticidal Agents

Background: Plant secondary metabolites play an essential role in the discovery of novel insecticide due to their unique sources and potential target sites. Paeonol, the main phenolic components in Moutan Cortex, is recognized as a safe and potent botanical insecticide to many insects. The structural modification of paeonol in this study into phenylsulfonylhydrazone derivatives is proved an effective approach for the development of novel insecticides, those derivatives being more toxic than paeonol. However, there have been no reports on the insecticidal activity of paeonol-based phenylsulfonylhydrazone derivatives in controlling Mythimna separata. Methods: We have been working to discover biorational natural products-based insecticides. Twelve novel paeonol-based phenylsulfonylhydrazone derivatives have been successfully prepared by structural modification of paeonol, and the insecticidal activity against M. separata by the leafdipping method at the concentration of 1 mg/mL has been evaluated. Results: Insecticidal activity revealed that out of 12 title compounds, derivatives 5c and 5f displayed the best against M. separate with the FMR both of 53.6% than toosendanin (FMR = 50.0%). Conclusion: The results suggested that for the paeonol-based phenylsulfonylhydrazone series derivatives, the proper substituent of arylsulfonyl R at the hydroxyl position of paeonol was very important for their insecticidal activity. These preliminary results will pave the way for further modification of paeonol in the development of potential new insecticides.

Paeonol, an Ingredient of Kamishoyosan, Reduces Intracellular Lipid Accumulation by Inhibiting Glucocorticoid Receptor Activity in 3T3-L1 Cells

Excessive triglyceride accumulation in lipid-metabolizing tissues is associated with an increased risk of a variety of metabolic diseases. Kamishoyosan (KSS) is a Kampo composed of 10 constituent herbs, and contains moutan cortex (MC) and paeonol (PN) as the major ingredient of MC. Here, we demonstrate the molecular mechanism underlying the effect of KSS on the differentiation of mouse preadipocytes (3T3-L1 cells). KSS inhibited the accumulation of triglycerides in a dose-dependent manner in 3T3-L1 cells that were induced to differentiate into adipocytes. We also found that MC and PN were responsible for the anti-adipogenetic effect of KSS and significantly suppressed the expression of CCAAT/enhancer-binding proteins-δ (C/EBP-δ) mRNA 3 days after the induction of differentiation. Thus, PN may contribute to the anti-adipogenetic property of MC in 3T3-L1 cells. In addition, PN inhibited dexamethasone (Dex)-induced glucocorticoid receptor (GR) promoter activity. Taken together, these results suggest that PN suppresses C/EBP-δ expression by inhibiting Dex-induced GR promoter activity at the early stage of differentiation and, consequently, delays differentiation into mature adipocytes. Our results suggest that the habitual intake of Kampo-containing PN contributes to the prevention of the onset of metabolic diseases by decreasing the excessive accumulation of triglycerides in lipid-metabolizing tissues.