Contamination with bacterial endotoxin causes the disruption of the tight junction (TJ) barrier. We investigated the ameliorative effect of dietary flavonoids genistein (Ge) and daidzein (Di) in normal or LPS-induced the disruption of epithelial barrier function of the endometrium. Using the immortalized porcine glandular endometrial epithelial cells (PEG), transepithelial electrical resistance (TER) and fluorescein isothiocyanate-dextran flux (FD-4) across the monolayer were measured. The mRNA expression of tight junction proteins, zona occludens-1 (ZO-1) and claudin-1, -3, -4, -7 and -8 was evaluated by real-time reverse transcription-polymerase chain reaction for coinciding effect of Ge or Di occurred at the gene transcription level. The results revealed that Ge and Di altered the TER depending on times and concentrations. Low concentration (10^-10 M) of both compounds decreased the TER whereas higher concentrations (10^-8 or ^-6 M) increased the TER which was not related to the FD-4 flux. The increased TER by Ge or Di was parallel to the induction of claudin-3 and -4 or -8 mRNA expression, respectively. With LPS inoculation, all isoflavone treatments inhibited the decreased TER induced by LPS, but only Ge (10^-8 or 10^-6 M) or Di (10^-10 or 10^-6 M) was coincidence with the decreased FD-4 flux. Under this LPS-stimulated condition, some or all examined TJ gene expressions appeared to be promoted by specific concentration of genistein or daidzein, respectively. Our findings suggest that the soy isoflavones treatment could promote and restore the impaired endometrial barrier function caused by LPS contamination.

The present study aimed to investigate the impact of Clostridium butyricum probiotic feed additive on sow and piglet performances, together with alterations in the lipidomic and metabolomic profiles of sow milk. Sixty-four Landrace × Yorkshire crossbred sows and 794 piglets were included. Sows were divided into two groups; i.e., (i) conventional gestation diet (control; n = 35) and (ii) conventional diet added with 10 g/sow/day of probiotic C. butyricum spores (treatment; n = 29) from one week before the estimated farrowing day until weaning (29.6 ± 4.8 days). The sow and piglet performances and incidence of piglet diarrhea were recorded. Changes in gross chemical composition, fatty acid and non-volatile polar metabolite profiles of sow colostrum, transient milk and mature milk were evaluated. The results showed that relative backfat loss in the treatment group (−2.3%) was significantly lower than in control group (11.6%), especially in primiparous sows (p = 0.019). The application of C. butyricum probiotics in sows significantly reduced the incidence of diarrhea in piglets (p < 0.001) but no other effect on piglet performance was found. Lipidomic and metabolomic analyses revealed variations in sow colostrum and milk biomolecular profiles, with indicative compounds significantly altered by feeding with the C. butyricum probiotics. In conclusion, the use of C. butyricum probiotics in sows may improve sow body condition and reduce diarrhea incidence in piglets, with underlying changes in milk composition that warrant further investigation. These findings support the potential of C. butyricum as a beneficial feed additive in swine production.

High ambient temperature (HTa) causes acid–base imbalance and systemic oxidative stress, and this may indirectly affect the mammary gland. Furthermore, HTa induces intracellular oxidative stress, which has been proposed to affect cell metabolism directly. We previously showed in dairy goats that the negative effect of HTa was compromised by enhancing heat dissipation during a high dietary cation and anion difference (DCAD) regimen. Moreover, high-dose vitamin C or ascorbic acid (AA) supplements have been used to manage oxidative stress in ruminants. The present study hypothesized that high DCAD and AA supplements that could alleviate the HTa effect would influence the milk synthesis pathway and mammary gland function. The results showed that goats fed with high DCAD had higher blood pH than control goats in the 4th week. The high dose of AA supplement decreases urine pH in the 8th week. The percent reduction of urine pH from the AA supplement was significant in the DCAD group. The high-dose AA supplement decreased plasma glutathione peroxidase activity and malonaldehyde. This effect was enhanced by a high DCAD supplement. In addition, supplementation with AA increased milk protein and citrate and decreased milk FFA. These alterations indicate the intracellular biochemical pathway of energy metabolism and milk synthesis. It can be concluded that a high DCAD regimen and AA supplement in dairy goats fed under HTa could influence the milk synthesis pathway. The evidence suggests that HTa decreases mammary gland function by modification of acid–base homeostasis and oxidative stress.

Kaempferol is a flavonol identified as the most potent activator of chloride (Cl⁻) secretion among other flavonoids in airway epithelial cells. This study aimed to investigate the cellular mechanisms by which kaempferol stimulates Cl⁻ secretion in the T84 human colon carcinoma cell line by Ussing chambers and voltage clamp technique. Bilateral addition of kaempferol (1–100 µM) increased short-circuit current (I_sc) in a concentration-dependent manner. Ion substitution of Cl⁻ or CFTR inhibitors NPPB and glibenclamide or a Na⁺/K⁺/2Cl⁻ cotransporter inhibitor bumetanide attenuated kaempferol-induced I_sc response. In permeabilized monolayers, selective channel inhibitors CFTRinh-172 and CaCCinh-A01 inhibited kaempferol-induced apical Cl⁻ current (I_Cl), and K⁺ blockers BaCl₂ and clotrimazole inhibited basolateral K⁺ current (I_Kb). The kaempferol-induced I_Cl showed no additive effects with forskolin or 8cpt-cAMP. The kaempferol-induced I_Cl was mostly abolished by protein kinase A inhibitor H89, but not by tyrosine kinase inhibitors, AG490 and tyrphostin A23, or tyrosine phosphatase inhibitor vanadate. Treatment with kaempferol for 24 h increased the expression of CFTR protein as determined by the Western blot analysis. These results demonstrated that kaempferol activates Cl⁻ secretion across T84 cells by activating the apical Cl⁻ current and basolateral K⁺ current. The mechanisms may involve the cAMP/PKA pathway and CFTR expression. Taken together, these findings reveal the beneficial effects of kaempferol to increase fluid secretion which can be used to treat constipation.

Modification of cellular and immunological events due to porcine reproductive and respiratory syndrome virus (PRRSV) infection is associated with pathogenesis in lungs. PRRSV also causes female reproductive dysfunction and persistent infection which can spread to fetus, stillbirth, and offspring. In this study, changes in cellular and innate immune responses to PRRSV type 1 or type 2 infection, including expression of PRRSV mediators, mRNA expression of Toll-like receptors (TLRs) and cytokine, and cytokine secretion, were examined in primary porcine glandular endometrial cells (PGE). Cell infectivity as observed by cytopathic effect (CPE), PRRSV nucleocapsid proteins, and viral nucleic acids was detected as early as two days post-infection (2 dpi) and persisted until 6 dpi. A higher percentage of CPE and PRRSV-positive cells were observed in type 2 infections. PRRSV mediator proteins, CD151, CD163, sialoadhesin (Sn), integrin and vimentin, were upregulated following type 1 and type 2 infection.CD151,CD163andSnwere upregulated by type 2. In both PRRSV types, mRNA expression of TLR1 and TLR6 was upregulated. However,TLR3was upregulated by type 1, butTLR4andTLR8mRNA and protein were downregulated by type 2 only.Interleukin(IL)-1β,IL-6andtumor necrotic factor(TNF)-αwere upregulated by type 2, butIL-8was upregulated by type 1. Both PRRSV type 1 and 2 stimulated IL-6 but suppressed TNF-α secretion. In addition, IL-1β secretion was suppressed only by type 2. These findings reveal an important mechanism underlying the strategy of PRRSV infection in the endometrium and associated with the viral persistence.