It is well-known that essential oil thymol exhibits antibacterial activity. The protective effects of thymol on pig intestine during inflammation is yet to be investigated. In this study, an in vitro ...lipopolysaccharide (LPS)-induced inflammation model using IPEC-J2 cells was established. Cells were pretreated with thymol for 1 h and then exposed to LPS for various assays. Interleukin 8 (IL-8) secretion, the mRNA abundance of cytokines, reactive oxygen species (ROS), nutrient transporters, and tight junction proteins was measured. The results showed that LPS stimulation increased IL-8 secretion, ROS production, and tumor necrosis factor alpha (TNF-α) mRNA abundance (P < 0.05), but the mRNA abundance of sodium-dependent glucose transporter 1 (SGLT1), excitatory amino acid transporter 1 (EAAC1), and H+/peptide cotransporter 1 (PepT1) were decreased (P < 0.05). Thymol blocked ROS production (P < 0.05) and tended to decrease the production of LPS-induced IL-8 secretion (P = 0.0766). The mRNA abundance of IL-8 and TNF-α was reduced by thymol pretreatment (P < 0.05), but thymol did not improve the gene expression of nutrient transporters (P > 0.05). The transepithelial electrical resistance (TEER) was reduced and cell permeability increased by LPS treatment (P < 0.05), but these effects were attenuated by thymol (P < 0.05). Moreover, thymol increased zonula occludens-1 (ZO-1) and actin staining in the cells. However, the mRNA abundance of ZO-1 and occludin-3 was not affected by either LPS or thymol treatments. These results indicated that thymol enhances barrier function and reduce ROS production and pro-inflammatory cytokine gene expression in the epithelial cells during inflammation. The regulation of barrier function by thymol and LPS may be at post-transcriptional or post-translational levels.
•The LC-QTOF/QqQ-MS was used to identify and quantify the phenolics in Tetrastigma hemsleyanum Leaves (THL) extract.•The anticancer effects and mechanism of THL were investigated at the first ...time.•THL induced apoptosis by extrinsic and intrinsic apoptosis pathways in HepG2 cells.•THL could inhibit the tumor growth in H22 tumor-bearing mice.•The phenolics of THL might play key role in anticancer effects.
Tetrastigma hemsleyanum leaves (THL), a popular anticancer functional food in China, are used as nutrient or dietary supplement. However, the anticancer effects and the underlying mechanism of THL are still unknown. Thus, the phenolic profiles and anticancer activities of THL purified extract were investigated by a combination of chemical assays, LC-QTOF-MS/LC-QqQ-MS techniques, cell cultures and tumor-bearing mice model. It has been indicated that the TPC/TFC (421.63 and 343.38 mg/g DW) of THL extract were obviously increased after SPE purification, and the main phenolic components were apigenin and luteolin glycosides. Besides, THL could induce apoptosis by both of extrinsic death receptor pathway and intrinsic mitochondria pathway in HepG2 cells as well as inhibit the tumor growth, regulate Bcl-2 family proteins, and active the caspase family proteins in H22 tumor-bearing mice. Summarily, the findings clearly demonstrated that THL could be a potential functional food for the treatment and prevention of liver cancer.
The extracellular calcium-sensing receptor (CaSR) and vitamin D receptor (VDR) play important roles in regulating calcium mobilization, calcium absorption, and calcium homeostasis, and they could be ...potential therapeutic targets to osteoporosis in laying hens. The present study investigated the molecular distribution of CaSR and VDR and the localization of CaSR in the kidney, proventriculus (true stomach), duodenum, jejunum, ileum, colon, cecum, shell gland, and tibia of laying hens at 3 different laying stages (19, 40, and 55 wk). The results showed that the relative mRNA abundance of CaSR in the kidney, ileum, proventriculus, duodenum, and colon was higher (P < 0.05) than the other tissues at 40 and 55 wk. The relative mRNA abundance of CaSR in the tibia was higher (P < 0.05) at 55 wk than at 40 wk. However, there were no significant differences in the relative protein abundance of CaSR among all tested tissues at peak production or in each tissue at the 3 different laying stages (P > 0.05). The relative mRNA abundance of VDR was higher (P < 0.05) in the small intestine (duodenum, jejunum, and ileum) when compared with other tissues at the 3 different laying stages. The relative protein abundance of VDR in the duodenum was higher (P < 0.05) than that in the proventriculus, colon, and cecum. There were no significant differences in the VDR expression among the tested tissues at the 3 different laying stages (P > 0.05). The immunohistochemical results showed that the positive staining was found widely in each tissue. Moreover, different laying stages did not affect the localization of CaSR except for the tibia tissue. In conclusion, similar to VDR, CaSR was widely expressed not only in the gut but also in the tibia and shell gland in laying hens. The expression level of CaSR and VDR in all tested tissues was unchanged at the different laying stages.
•Ca2+ increased ITCs by up-regulating gene expression on GL synthesis and hydrolysis.•Ca2+ increased ITCs by increasing MYR activity.•Ca2+ increased the relative content of 1-isothiocyanato-butane in ...ITCs.•ITCs contributed to total antioxidant capacity and DPPH• scavenging capacity.
The effects of CaCl2 on metabolism of glucosinolates (GLs) and formation of isothiocyanates (ITCs), as well as the antioxidant capacity of broccoli sprouts, were investigated. The results showed that CaCl2 treatment enhanced BrST5b (sulfotransferase 5b) and limited AOP2 (2-oxoglutarate-dependent dioxygenase 2) expression, which increased GLs biosynthesis. CaCl2 also promoted the gene expression of myrosinase (MYR) and its activity, and thus increased ITCs formation. Three ITCs and two nitriles were found in the hydrolysate of broccoli sprouts. CaCl2 treatment increased the content of 1-isothiocyanato butane significantly. Then, CaCl2 increased the formation of ITCs by regulating gene expression and enzyme activity. After CaCl2 treatment, the antioxidant capacity of sprouts increased and the formed ITCs mainly contributed to total antioxidant capacity and DPPH• scavenging capacity.
We investigated efficacy of in ovo application of epidermal growth factor (EGF) on intestinal expression of EGF receptor (EGFR) during embryogenesis (experiment 1) and posthatch growth performance ...and gastrointestinal development in broiler chickens (experiment 2). In experiment 1, 450 fertile Ross 708 eggs were allocated to 3 groups (150 eggs/group): 1) control, 2) 160 μg EGF/kg of egg, and 3) 640 μg of EGF/kg of egg. Eggs were candled for live embryos on day 16 and injected with the respective treatment solutions on day 17 and sampled for jejunal tissue from day 17 to hatch for EGFR analyses. There was no effect of EGF (P > 0.05) on EGFR expression on day 17 to 20; however, on day 21, EGF increased (P < 0.05) EGFR expression in EGF birds relative to control birds. In experiment 2, 600 fertile Ross 708 eggs were allocated to 5 treatments: 1) intact, no puncture or injection, 2) punched but not injected, 3) control, no EGF, 4) 80 μg of EGF/kg of egg, and 5) 160 μg of EGF/kg of egg. The eggs were incubated and candled for live embryos on D 19, treated, and subsequently transferred to the hatcher. Upon hatching, chicks were weighed, and 90 chicks per treatment placed in cages (15 birds/cage) and allowed free access to a standard antibiotic-free corn-soybean diet for 21 D. Feed intake and body weight were monitored on a weekly basis. Samples of birds were necropsied on D 0, 7, 14, and 21 for measurements of intestinal weight and jejunal histomorphology and excreta samples taken on D 3 to 5 and 17 to 19 for apparent retention of dry matter. There was no EGF effect (P > 0.05) on any posthatch response criteria. In conclusion, in ovo application of EGF increased EGFR expression but had no effect on posthatch growth performance, DM retention, and intestinal development. The lack of EGF effect on posthatch response was surprising but suggested in ovo application of EGF may not be a viable approach.
Red-osier dogwood, a native species of flowering plant in North America, has been reported to have anti-oxidative properties because of abundant phenolic compounds; this could be promising as a ...functional food or a feed additive. In the present study, an oxidative damage model using 1.0 mM hydrogen peroxide (H2O2) in Caco-2 cells was established to evaluate the antioxidative effects of red-osier dogwood extracts (RDE). The results showed that 1.0 mM H2O2 pre-exposure for 3 h significantly decreased cell viability, and increased interleukin 8 (IL-8) secretion and the intracellular reactive oxygen species (ROS) level. Caco-2 cells were treated with 100 µg/mL RDE for 24 h after pre-exposure to H2O2. It was found that the decreased cell viability caused by H2O2 was significantly restored by a subsequent 100 µg/mL RDE treatment. Furthermore, the IL-8 secretion and ROS level were significantly blocked by RDE, accompanied by the enhanced gene expression of hemeoxygenase-1 (HO-1), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px), and the enhanced protein expression of the nuclear factor (erythroid-derived 2)-like 2 (Nrf-2). Moreover, RDE improved barrier functions in Caco-2 cells. Using RDE reduced the diffusion of fluorescein isothiocyanate (FITC)-dextran and increased the transepithelial resistance (TEER) value. The relative mRNA level of tight junction claudin-1, claudin-3, and occludin was elevated by RDE. These extracts also repaired the integrity of zonula occludens-1 (ZO-1) damaged by H2O2 and increased the protein expressions of ZO-1 and claudin-3 in the H2O2-pretreated cells. These results illustrated that RDE reduced the ROS level and enhanced the barrier function in oxidative-damaged epithelial cells.
Red-osier dogwood extracts (RDE) contain high levels of phenolic compounds which have been recognized as natural antioxidants. In this study, the potential of RDE to prevent cardiovascular diseases ...(CVDs) was evaluated using Caco-2 cells and a co-culture model of Caco-2 BBe1/EA.hy926 cells in Transwell® plates. The results showed that RDE supplementation significantly prevented interleukin-8 (IL-8) production and suppressed the gene expression of IL-8, tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and cyclooxygenase 2 (COX-2) in the TNF-α inflamed Caco-2 cells. Meanwhile, the polyphenols (quercetin-3-glucoside, quercetin-glucuronide, rutin, quercetin-3-O-malonylglucoside, and kaempferol-glucoside) in the RDE were validated to be absorbed by Caco-2 BBe1 cells and transported to the basal chamber where EA.hy926 cells were located during 12 h incubation. The transported polyphenols were able to prevent IL-8 production and suppress the gene expression of proinflammatory mediators (TNF-α, ICAM-1, VCAM-1, and COX-2) in the TNF-α or oxidized low-density lipoprotein (ox-LDL) treated EA.hy926 cells. These novel findings demonstrated that phenolic compounds in RDE can be transported to the cardiovascular system by intestinal absorption and mitigate the inflammatory responses of vascular endothelial cells, indicating that RDE could be a natural resource of polyphenols to prevent inflammation cytokine or oxidized lipid-induced CVDs.
In this study, sugary maize dendrimer-like glucan (SMDG) was used as a delivery carrier for improving the bioavailability of resveratrol (RES). After optimization, the solubility of RES in RES-SMDG ...markedly increased to approximately 9.1 times that of the raw RES solution. The structural characterizations of the RES-SMDG formulation showed crystal RES was entrapped in the SMDG matrix for the amorphous state due to the strong intermolecular hydrogen bonds between the -OH of RES and glucan chains. In this case, antioxidant activity of RES-SMDG was markedly higher than that of the raw RES solution. In the Caco-2 cell model, the
value of RES in the RES-SMDG group was slightly higher than those of common permeable compounds, while the cellular uptake was significantly improved. RES-SMDG also exhibited protective effects against cellular damage under oxidative stress. The results indicated that SMDG is an attractive carrier to encapsulate and protect hydrophilic bioactive ingredients.
•Dendrimer-like glucan nanoparticulate system (CoQ10-SMDG) was formulated using solid-dispersion preparation.•CoQ10-SMDG improved the water solubility of CoQ10 for 188.8-folds.•Structural ...characterizations revealed that crystal CoQ10 was entrapped in SMDG matrix for amorphous state.•The cellular antioxidant activity of CoQ10-SMDG was significantly improved compared with raw CoQ10.•SMDG was an excellent platform for encapsulation and delivery of hydropholic components.
Aqueous coenzyme Q10 (CoQ10) dispersions were prepared using sugary maize dendrimer-like glucan (SMDG) with solid-dispersion treatment. After measuring solubility, recovery rate and loading rate, the initial weight ratio of CoQ10:SMDG was optimized to be 1:27, with the solubility markedly increasing up 188.8-folds compared to pure CoQ10 solution. The structural characterizations of CoQ10-SMDG formulation showed crystal CoQ10 was entrapped in SMDG matrix for amorphous state, associated with the strong interactions with glucan chains. The antioxidant activity of CoQ10-SMDG was assessed via DPPH and FRAP assay. DPPH scavenging activity and FRAP value of it were as high as 95.1% and 0.87 mM, respectively. The cellular uptake of CoQ10 in CoQ10-SMDG group was significantly higher than that of natural CoQ10. CoQ10-SMDG also exhibited significant protective effects against cellular damage in H2O2-induced HaCaT cell model. The results indicated that dendrimer-like glucan is an excellent platform to encapsulate and improve biological activity of hydropholic compounds.
Poultry is vulnerable to bone problems throughout their lives or production period due to rapid growth in broilers and the active laying cycle in layers. The calcium-sensing receptor (CaSR) is ...important in calcium and bone metabolism. The objective of this study was to investigate the effect of the CaSR ligand (l-Trp) and 1,25-dihydroxycholecalciferol (1,25OHD3) on the regulation of proliferation and osteogenic differentiation of chicken mesenchymal stem cells (MSCs) isolated from the compact bones of 14-day-old Ross 308 chickens and Dekalb pullets, which can provide cell-based evidence for the prevention or alleviation of skeletal disorders in the poultry industry. First, the dose- (0, 0.5, 1, 2, 5, 10, and 15 mM) and time-effects (0, 7, and 14 days) of l-Trp on the proliferation and osteogenic differentiation in chicken MSCs were investigated. The 5 mM l-Trp had a balanced effect between proliferation and osteogenic differentiation in broiler and layer MSCs when differentiated for 7 days. The broiler and layer MSCs were then treated with (1) osteogenic medium, osteogenic medium supplemented with (2) 1 nM 1,25OHD3, (3) 2.5 mM Ca2+, (4) 2.5 mM Ca2+ + 5 mM l-Trp and (5) 2.5 mM Ca2+ + 5 mM l-Trp + 1 μM NPS-2143, separately for 7 days. Results showed that the 5 mM l-Trp significantly inhibited the proliferation of broiler and layer MSCs on day 7 (P < 0.05), but 1 nM 1,25OHD3 significantly promoted the proliferation of layer MSCs (P < 0.05). Only the 2.5 mM Ca2+ + 5 mM l-Trp group significantly increased the mineralization process during osteogenic differentiation (P < 0.05), and this treatment also significantly upregulated the mRNA expression of the vitamin D receptor (VDR), β-catenin, and osteogenesis genes in broiler MSCs (P < 0.05). The osteogenic differentiation process in layer MSCs was faster than that in broiler MSCs. In layer MSCs, Ca2+ alone significantly facilitated mineralization and ALP activity after 7-day osteogenic differentiation (P < 0.05). However, the 5 mM l-Trp significantly inhibited the differentiation and mineralization process by downregulating the mRNA expression of CaSR, VDR, β-catenin, and osteogenic genes (P < 0.05) in layer MSCs. Taken together, l-Trp and 1,25OHD3 can regulate proliferation and osteogenic differentiation in both broiler and layer MSCs depending on the dose, treatment time, and cell proliferation and differentiation stages.
PDF
