Effect and mechanism of Magnolia officinalis pharmacopuncture for treating localized fat via network pharmacology and experimental study.

Background: Recently, for various reasons, the need for non-invasive treatment for localized fat has emerged. This study confirmed whether Magnolia officinalis (MO) pharmacopuncture reduces localized fat by promoting lipolysis and inhibiting adipogenesis. Methods: The network was built using genes related to the active compound of MO and the mode of action of MO was predicted by the functional enrichment analysis. Based on the result from network analysis, 100 µL of 2 mg/mL MO pharmacopuncture was injected into the inguinal fat pad for 6 weeks in obese C57BL/6J mice. Normal saline was injected into the right-side inguinal fat pad as a self-control. Results: It was expected that the 'AMP-activated protein kinase (AMPK) signaling pathway' would be affected by the MO Network. MO pharmacopuncture reduced the weight and size of inguinal fat in HFD-induced obese mice. The phosphorylation of AMPK along with the increases of lipases was significantly increased by MO injection. Also, the expression levels of fatty acid synthesize-related mediators were suppressed by MO injection. Conclusion: Our results demonstrated that MO pharmacopuncture promoted the expression of AMPK, which has beneficial effects on activation of lipolysis and inhibition of lipogenesis. Pharmacopuncture of MO can be a non-surgical alternative therapy in the treatment of local fat tissue.

Network Pharmacology and Molecular Docking Study on the Multi-Target Mechanisms of Aloe vera for Non-Alcoholic Steatohepatitis Treatment.

Non-alcoholic steatohepatitis (NASH) is a leading cause of chronic liver disease with limited treatment options. The widely distributed plant Aloe vera has shown protective effects against NASH in animals, yet the precise mechanism remains unknown. In this study, we investigated the potential mechanisms underlying the anti-NASH effects of Aloe vera using a network pharmacology and molecular docking approach. By searching online databases and analyzing the Gene Expression Omnibus dataset, we obtained 260 Aloe vera-NASH common targets. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses showed that the common targets were strongly associated with the key pathological processes implicated in NASH, including lipid and glucose metabolism, inflammation, apoptosis, oxidative stress, and liver fibrosis. Four core proteins, AKT serine/threonine kinase 1 (AKT1), tumor necrosis factor alpha (TNFα), transcription factor c-Jun, and tumor suppressor protein p53, were identified from compound-target-pathway and protein-protein interaction networks. Molecular docking analysis verified that the active ingredients of Aloe vera were able to interact with the core proteins, especially AKT1 and TNFα. The results demonstrate the multi-compound, multi-target, and multi-pathway mechanisms of Aloe vera against NASH. Our study has shown the scientific basis for further experiments in terms of the mechanism to develop Aloe vera-based natural products as complementary treatments for NASH. Furthermore, it identifies novel drug candidates based on the structures of Aloe vera's active compounds.

Research Note: Improvement of eggshell strength and intensity of brown eggshell color by dietary magnesium and δ-aminolevulinic acid supplementation in laying hens.

The objective of the current experiment was to investigate the effect of dietary magnesium (Mg) and δ-aminolevulinic acid (ALA) supplementation on productive performance and egg quality in laying hens. The present experiment was performed using a completely randomized design with 2 × 2 factorial arrangements including 2 levels of supplemental Mg (0 and 0.12%) and ALA (0 and 0.25%) in diets. A total of 192 43-wk-old Hy-Line Brown laying hens were allotted to 1 of 4 dietary treatments with 6 replicates. Each replicate consisted of 4 consecutive cages with 2 hens per cage. Treatment diets were fed to hens for 6 wk. Results indicated that dietary Mg supplementation improved (P < 0.01) eggshell strength but decreased (P < 0.05) eggshell color score, regardless of dietary ALA supplementation. However, the decreased eggshell color by dietary Mg supplementation was relieved by dietary ALA supplementation, which led to a significant interaction (P < 0.05). Likewise, an interaction (P < 0.05) was also observed for eggshell lightness (L*) because dietary ALA supplementation decreased eggshell L* values only when Mg was supplemented to diets. In conclusion, dietary Mg supplementation improves eggshell strength but decreases the intensity of brown eggshell color in laying hens. However, this negative effect of dietary Mg supplementation on eggshell color is likely ameliorated by dietary ALA supplementation.

Effects of stocking density and dietary vitamin C on performance, meat quality, intestinal permeability, and stress indicators in broiler chickens.

The objective of the current study was to investigate the effects of stocking density (SD) and dietary supplementation of vitamin C on growth performance, meat quality, intestinal permeability, and stress indicators in broiler chickens. The study was conducted using a completely randomized design with a 2 × 2 factorial arrangement consisting of 2 different SD and 2 supplemental levels of dietary vitamin C. A total of 1,368 Ross 308 broiler chickens of 21 days of age with similar body weights (BW) were randomly allotted to 1 of 4 treatments with 6 replicates each. Different numbers of birds per identical floor pen (2.0 m × 2.4 m) were used to create 2 different SD levels of low SD (9 birds/m2) and high SD (18 birds/m2). The basal diet was formulated with no supplemental vitamin C to meet or exceed nutrient recommendations of the Ross 308 manual. The other diet was prepared by supplementing 200 mg/kg vitamin C in the basal diet. The study lasted for 14 days. At the end of the study, 3 male birds per replicate were selected to analyze meat quality, intestinal permeability, and stress indicators such as blood heterophil:lymphocyte (H:L) and feather corticosterone (CORT) concentrations. Results indicated that there were no interactions between different SD and dietary supplementation of vitamin C for all measurements. For the main effects of SD, birds raised at high SD had less (p < 0.01) BW, BW gain, and feed intake with increasing stress responses including greater blood H:L and feather CORT concentrations (p < 0.01) than those raised at low SD. Transepithelial electrical resistance in the jejunal mucosa was decreased (p < 0.05) at high SD, indicating an increase in intestinal permeability. However, the main effects of dietary supplementation of 200 mg/kg vitamin C were insignificant for all measurements. In conclusion, high SD of broiler chickens impairs growth performance and intestinal barrier function with increasing stress responses. However, dietary supplementation of vitamin C may have little beneficial effects on broiler chickens raised at the high SD condition used in the present study.

Comparative effects of dietary functional nutrients on growth performance, meat quality, immune responses, and stress biomarkers in broiler chickens raised under heat stress conditions.

OBJECTIVE: The objective of the present study was to investigate the comparative effects of dietary functional nutrients including glutamine (Gln), chromium picolinate (Cr picolinate), vitamin C (Vit C), betaine (Bet), and taurine (Tau) on growth performance, meat quality, immune responses, and stress biomarkers in broiler chickens raised under heat stress conditions. METHODS: A total of 420 21-d-old Ross 308 male broiler chickens (initial body weight = 866±61.9 g) were randomly allotted to 1 of 7 treatment groups with 6 replicates. One group was kept under thermoneutral conditions and was fed a basal diet (PC, positive control). Other 6 groups were exposed to a cyclic heat stress condition. One of the 6 groups was fed the basal diet (NC, negative control), whereas 5 other groups were fed the basal diet supplemented with 0.5% Gln, 500 ppb Cr picolinate, 250 mg/kg Vit C, 0.2% Bet, or 1.0% Tau. The diets and water were provided ad libitum for 21 d. RESULTS: Broiler chickens in NC group had decreased (p<0.05) growth performance and immune responses measured based on cutaneous basophil hypersensitivity (CBH), but increased (p<0.05) stress responses measured based on feather corticosterone concentrations and blood heterophil:lymphocyte than those in PC group. However, none of dietary functional nutrients had a positive effect on growth performance of broiler chickens. Dietary supplementation of 250 mg/kg Vit C improved (p<0.05) CBH responses of broiler chickens, but other functional nutrients had no such an improvement in CBH responses. All functional nutrients decreased (p<0.05) stress responses of broiler chickens. CONCLUSION: Functional nutrients including Gln, Cr picolinate, Vit C, Bet, and Tau at the supplemental levels used in this study decrease stress responses of broiler chickens to a relatively similar extent. However, this reduction in stress responses could not fully ameliorate decreased productive performance of broiler chickens raised under the current heat stress conditions.

Pharmacokinetic Properties of Moracin C in Mice.

Moracin C from Morus alba fruits, also known as the mulberry, has been proven to exhibit inhibitory activities against lipoxygenase enzymes, TNF-α and interleukin-1ß secretion, and proprotein convertase subtilisin/kexin type 9 expression. Despite the various pharmacological activities of moracin C, its pharmacokinetic characteristics have yet to be reported. Here, the pharmacokinetic parameters and tissue distribution of moracin C have been investigated in mice, and the plasma concentration of moracin C with multiple dosage regimens was simulated via pharmacokinetic modeling. Our results showed that moracin C was rapidly and well absorbed in the intestinal tract, and was highly distributed in the gastrointestinal tract, liver, kidneys, and lungs. Moracin C was distributed in the ileum, cecum, colon, and liver at a relatively high concentration compared with its plasma concentration. It was extensively metabolized in the liver and intestine, and its glucuronidated metabolites were proposed. In addition, the simulated plasma concentrations of moracin C upon multiple treatments (i.e., every 12 and 24 h) were suggested. We suggest that the pharmacokinetic characteristics of moracin C would be helpful to select a disease model for in vivo evaluation. The simulated moracin C concentrations under various dosage regimens also provide helpful knowledge to support its pharmacological effect.

Cardamonin Inhibits Oxazolone-Induced Atopic Dermatitis by the Induction of NRF2 and the Inhibition of Th2 Cytokine Production.

The skin is constantly exposed to various types of chemical stresses that challenge the immune cells, leading to the activation of T cell-mediated hypersensitivity reactions including atopic dermatitis. Previous studies have demonstrated that a variety of natural compounds are effective against development of atopic dermatitis by modulating immune responses. Cardamonin is a natural compound abundantly found in cardamom spices and many other medicinal plant species. In the present study, we attempted to examine whether cardamonin could inhibit oxazolone-induced atopic dermatitis in vivo. Our results show that topical application of cardamonin onto the ear of mice suppressed oxazolone-induced inflammation in the ear and hyperplasia in the spleen. Cardamonin also inhibited oxazolone-induced destruction of connective tissues and subsequent infiltration of mast cells into the skin. In addition, we found that the production of Th2 cytokines is negatively regulated by NRF2, and the induction of NRF2 by cardamonin contributed to suppressing oxazolone-induced Th2 cytokine production and oxidative damages in vivo. Together, our results demonstrate that cardamonin is a promising natural compound, which might be effective for treatment of atopic dermatitis.

Discovery of Flavonoids from Scutellaria baicalensis with Inhibitory Activity Against PCSK 9 Expression: Isolation, Synthesis and Their Biological Evaluation.

Nine flavonoids were isolated and identified from a chloroform-soluble fraction of the roots of Scutellaria baicalensis through a bioactivity-guided fractionation using a proprotein convertase subtilisin/kexin type 9 (PCSK9) monitoring assay in HepG2 cells. All structures were established by interpreting the corresponding spectroscopic data and comparing measured values from those in the literature. All compounds were assessed for their ability to inhibit PCSK9 mRNA expression; compounds 1 (3,7,2'-trihydroxy-5-methoxy-flavanone) and 4 (skullcapflavone II) were found to suppress PCSK9 mRNA via SREBP-1. Furthermore, compound 1 was found to increase low-density lipoprotein receptor protein expression. Also, synthesis of compound 1 as a racemic mixture form (1a) was completed for the first time. Natural compound 1 and synthetic racemic 1a were evaluated for their inhibitory activities against PCSK9 mRNA expression and the results confirmed the stereochemistry of 1 was important.

Design and synthesis of novel 2',3'-dideoxy-4'-selenonucleosides as potential antiviral agents.

On the basis of potent anti-HIV activity of 2',3'-dideoxynucleosides (ddNs), their bioisosteric analogues, 2',3'-dideoxy-4'-selenonucleosides (4'-seleno-ddNs) were first synthesized from a chiral template, d-glutamic acid using stereoselective ring-closure reaction of the dimesylate with Se(2-) and Pummerer type condensation of the selenoxide with nucleobases as key steps. X-ray crystallographic analysis indicated that 4'-seleno-ddNs adopted the same C2'-endo/C3'-exo (South) conformation as anti-HIV active ddNs, but did not show anti-HIV activity, indicating that RT seems to prefer the C2'-exo/C3'-endo (North) conformation on binding with their triphosphates.

Stereoselective synthesis and conformational study of novel 2',3'-Didehydro-2',3'-dideoxy-4'-selenonucleosides.

Stereoselective synthesis of novel 2',3'-didehydro-2',3'-dideoxy-4'-selenonucleosides (4'-seleno-d4Ns) 4a- c was accomplished via 4'-selenoribofuranosyl pyrimidines 11a- c, as key intermediates. 4'-Selenoribofuranosyl pyrimidines 11a- c were efficiently synthesized from d-ribose or d-gulonic gamma-lactone using a Pummerer-type condensation as a key step. Introduction of 2',3'-double bond was achieved by treating cyclic 2',3'-thiocarbonate with 1,3-dimethyl-2-phenyl-1,3,2-diazaphospholidine.

Oxidative stress and tumor necrosis factor-alpha-induced alterations in metaphase II mouse oocyte spindle structure.

OBJECTIVE(S): To examine the effect of exogenous exposure to hydrogen peroxide (H(2)O(2)) and tumor necrosis factor (TNF)-alpha on mouse metaphase II (MII) oocyte spindle structure and to examine the potential benefits of supplementing the culture media with vitamin C. DESIGN: Prospective study. SETTING: Research laboratory in a tertiary hospital. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Microtubule changes and alterations in chromosomal alignment. RESULT(S): Both concentration- and time-dependent alterations were seen in spindle structure after exposure to H(2)O(2). An H(2)O(2) concentration as low as 12.5 microM increased the odds of an oocyte with altered microtubule and chromosome alignment (score >or=3) by 93%. Significantly increased damage was seen with increasing period of incubation. Higher scores were seen after exposure to both TNF-alpha alone and in combination with H(2)O(2) compared with controls. Changes in chromosomal alignment were comparable among the three groups. Oocytes coincubated with H(2)O(2) and vitamin C at 200 microM demonstrated less damage compared with those with H(2)O(2) alone. CONCLUSION(S): Oxidative stress results in concentration and time-dependent alterations in the spindle structure and augments the effects induced by TNF-alpha. Proper oocyte handling in vitro may help reduce oxidative insult, thus improving the oocyte quality. Antioxidants may have a protective effect and need to be further evaluated.