Fermentation Improves Calcium Bioavailability in Moringa oleifera leaves and Prevents Bone Loss in Calcium-deficient Rats.

Nowadays, there is an increasing demand of healthier plant calcium supplements. Moringa oleifera leaves (MOL) are rich in calcium and thus are promising candidates for developing efficient calcium supplements. Here, using fermentation-based approaches, we developed a Moringa oleifera leaf ferment (MOLF), which contents higher levels of calcium. The therapeutic potential of the MOLF was also examined both in vitro and in vivo. Nine lactic acid bacteria and four yeasts were tested for better fermentation of MOL. Calcium-deficient rats were used for evaluating the therapeutic effects of MOLF. The results of liquid fermentation showed that the mixture of Lactobacillus reuteri, Lactobacillus acidophilus , and Candida utilis elevated the content of MOL calcium most strikingly, with the content of calcium increased nearly 2.4-fold (from 2.08% to 4.90%). The resulting MOLF was then subjected to cell experiments and animal experiments. The results showed that calcium absorption in Caco-2 cells in MOLF group was higher than that in CaCl2 group significantly. Interestingly, in calcium-deficient rats, MOLF treatment significantly increased the thickness of cortical bone, rat body weight, wet weight of the femur, and the femur bone density, whereas it decreased osteoclast numbers. These results indicate that microbial fermentation increased calcium bioavailability of MOL, promote the growth and development of calcium-deficient rats, bone calcium deposition, and bone growth; enhance bone strength; reduce bone resorption; and prevent calcium deficiency.

Melatonin inhibits endoplasmic reticulum stress-associated TXNIP/NLRP3 inflammasome activation in lipopolysaccharide-induced endometritis in mice.

Endometritis, an inflammatory response of the uterus tissue, is characterized by the production of inflammatory cytokines and migration of neutrophil (PMN) into the uterus tissue. Melatonin has been demonstrated to have anti-inflammatory and antioxidant effects. The purpose of this study was to investigate the protective effects of melatonin on lipopolysaccharide (LPS)-induced endometritis in mice. An endometritis model was induced by LPS and melatonin was given 1 h before LPS treatment. The results showed that melatonin inhibited LPS-induced pathologic changes, Myeloperoxidase (MPO) activity, and levels of interleukin-1 beta (IL-1ß). Melatonin also inhibited LPS-induced thioredoxin-interacting protein (TXNIP)/NOD-like receptor family, pyrin domain containing 3 (NLRP3) inflammasome and nuclear factor kappa B (NF-κB) activation, reactive oxygen species (ROS) production, and endoplasmic reticulum (ER) stress. Furthermore, melatonin was found to increase AMPK activity. In conclusion, our results demonstrated that melatonin inhibited ER stress-associated TXNIP/NLRP3 inflammasome activation with a regulation of adenosine monophosphate activated protein kinase (AMPK) in LPS-induced endometritis. Melatonin may serve as a promising nutritional supplement for the treatment of endometritis.

Farrerol regulates antimicrobial peptide expression and reduces Staphylococcus aureus internalization into bovine mammary epithelial cells.

Mastitis, defined as inflammation of the mammary gland, is an infectious disease with a major economic influence on dairy industry. Staphylococcus aureus is a common gram-positive pathogen that frequently causes subclinical, chronic infection of the mammary gland in dairy cows. Farrerol, a traditional Chinese medicine isolated from rhododendron, has been shown to have anti-bacterial activity. However, the effect of farrerol on S. aureus infection in mammary epithelium has not been studied in detail. The aim of this study was to investigate the effect of farrerol on the invasion of bovine mammary epithelial cells (bMEC) by S. aureus. The expression of antimicrobial peptide genes by bMEC were assessed in the presence or absence of S. aureus infection. Our results demonstrated that farrerol (4-16 µg/ml) reduced > 55% the internalization of S. aureus into bMEC. We also found that farrerol was able to down-regulate the mRNA expression of tracheal antimicrobial peptide (TAP) and bovine neutrophil ß-defensin 5 (BNBD5) in bMEC infected with S. aureus. The Nitric oxide (NO) production of bMEC after S. aureus stimulation was decreased by farrerol treatment. Furthermore, farrerol treatment suppressed S. aureus-induced NF-κB activation in bMEC. These results demonstrated that farrerol modulated TAP and BNBD5 gene expression in mammary gland, enhances bMEC defense against S. aureus infection and could be useful in protection against bovine mastitis.

Prevention of acute GVHD in mice by treatment with Tripterygium hypoglaucum Hutch combined with cyclosporin A.

OBJECTIVE: To elucidate the protective roles and the underlying mechanism of Tripterygium hypoglaucum Hutch (THH) in mice graft-versus-host disease (GVHD). METHODS: BALB/c (H-2k(d)) mice were firstly treated with total body irradiation and infused with a mixture of bone marrow and spleen cells from C57BL/6. Then the severity of acute GVHD (aGVHD), chimeras of donor cells, inflammatory cytokines (IFN-γ, IL-4, and IL-10) of plasma, and regulatory T cells were evaluated to elucidate the different drug combinations and concentrations of cyclosporin A (CsA) and THH in preventing aGVHD. RESULTS: The control group treated with phosphate buffer solution displayed more obvious ruffled hair, hunched posture, diarrhea, reduced weight and more lymphocytes infiltration into the spleen and intestine than these treated with CsA, THH or low-dosed CsA combined with THH, especially those treated with low-dosed CsA combined with THH. No significant differences were observed in the chimeras of donor cells and survival rate among the CsA, THH, or CsA combined with THH-treated groups. Further studies implied that THH might reduce the aGVHD by increasing IL-10, decreasing IL-4, activating Treg cell, and maintaining a relatively high Foxp3 mRNA level. CONCLUSION: THH decreased the occurrence of mouse aGVHD and prolonged the survival time by increasing the levels of CD(4)(+)/CD(25)(+) T cells, regulating the cytokine secretion and promoting the expression of Foxp3.

Salidroside attenuates inflammatory responses by suppressing nuclear factor-κB and mitogen activated protein kinases activation in lipopolysaccharide-induced mastitis in mice.

BACKGROUND AND OBJECTIVE: Mastitis is defined as inflammation of the mammary gland in domestic dairy animals and humans. Salidroside, a major component isolated from Rhodiola rosea L., has potent anti-inflammatory properties, but whether it can be used in mastitis treatment has not yet been investigated. The aim of this study was to assess the protective effects of salidroside against lipopolysaccharide (LPS)-induced mastitis in mice and the mechanism of action. METHODS AND RESULTS: We used a mouse mastitis model in which mammary gland inflammation was induced by LPS challenge. Salidroside administered 1 h before LPS infusion significantly attenuated inflammatory cell infiltration, reduced the activity of myeloperoxidase in mammary tissue, and decreased the concentration of tumor necrosis factor-α, interleukin (IL)-1ß, and IL-6 in a dose-dependent manner. Further studies revealed that salidroside down-regulated phosphorylation of LPS-induced nuclear transcription factor-kappaB (NF-κB) p65 and inhibitor of NF-κB α (IκBα) in the NF-κB signal pathway, and suppressed phosphorylation of p38, extracellular signal-regulated kinase (ERK) and c-jun NH(2)-terminal kinase (JNK) in MAPKs signal pathways. CONCLUSIONS: This study demonstrates that salidroside is an effective suppressor of inflammation and may be a candidate for the prophylaxis of mastitis.

[Experimental study of Tripterygium hypoglaucum (level) Hutch on preventing acute graft-versus-host disease in bone marrow transplantation mice].

OBJECTIVE: To evaluate the effects of Tripterygium hypoglaucum (level) Hutch (THH) on cytokine production in acute graft-versus-host disease (aGVHD)mice and explore the mechanisms. METHODS: 2 x 10(7) bone marrow cells mixed with 2 x 10(7) spleen cells from the same C57BL/6 mouse were transplanted into the myeloablative irradiated inbred BALB/c mouse to establish a aGVHD model. The experiments were designed as follows: control group (group A), CsA prophylaxis group (group B), THH prophylaxis group (group C), and combined THH with CsA prophylaxis group (group D). aGVHD was assessed by histologic changes of skin, liver and intestines. Chimerism was detected by H-2b molecular expression on recipient mice bone marrow cells with flow cytometer. Serum concentrations of IFN-gamma, IL-4 and IL-10 were determined by ELISA. RESULTS: The serum concentrations of IFN-gamma in group B, C, D were significantly lower than that in group A, while those of IL-10 was significantly higher than that in group A (P < 0.05). There was no changes in concentration of IL-4 in all the groups (P > 0.05). The median survival time for group A was nine days, while that of group B, C, D each was more than 30 days being significantly longer than that of group A (P < 0.05). The recipient mice of group A displayed significant clinical symptoms of GVHD, and died within 20 days; whereas those of group B, C, D showed only ruffled fur and uplift posture. Histologic changes of liver and intestines in group B and C displayed a few lymphocytes infiltration while the histologic morphology of skin, liver and intestines in the survived mice of group D was normal. Allogeneic chimerism rates of group B, C, D at day 30 after allo-BMT were (99.18 +/- 0.58)%, (97.68 +/- 0.59)%, and (99.15 +/- 0.11)%, respectively. CONCLUSION: THH could regulate the production of cytokines and prevent aGVHD. THH and CsA at low dose combination showed synergic effect in preventing aGVHD.