Enhanced anti-inflammatory effect of Rosmarinic acid by encapsulation and combination with the exosome in mice with LPS-induced endometritis through suppressing the TLR4-NLRP3 signaling pathway.

The TLR4-NLRP3 signaling pathway plays an essential role in the development of inflammation and especially endometritis. Rosmarinic acid (RA) can have potent anti-inflammatory effects in the drug-loading system. The purpose of this was to evaluate the anti-inflammatory effects of RA loaded to exosomes (RLE) on lipopolysaccharide (LPS)-induced endometritis in mice. RA was loaded into serum-derived exosome, using sonication methods. Animals in the treatment groups were subjected to uterine horn injection of RA, exosome, RA combination with exosome (R+E), and RA loaded to exosome (RLE) in uterine horn by two dosages in each group (5 and 10 mg/kg of RA or exosome), 24 h after inducing endometritis. Histopathological analysis, MPO production, immunohistochemistry, and qPCR were used to determine whether the treatment groups were adequate in controlling inflammation. The results showed that treatment groups, and mainly RLE10 and R10 +E10 groups, could modulate pathological changes, inhibit myeloperoxidase (MPO) activity, and significantly reduce the gene and protein expression of TLR4, NLRP3, inflammatory cytokines such as IL-1ß, IL-18, and TNF-α, and lastly, GSDM-D as a pyroptosis factor. In conclusion, RA loaded and combination with exosomes at a dosage of 10 mg/kg (RLE10 and R10 +E10) improved endometritis in mice through a suppressing TLR4-NLRP3 signaling pathway.

Effect of PGF2α and GnRH administration on reproductive performance in Ghezel ewes.

The main aim of the current study was to evaluate the effect of GnRH administration on day five after mating as well as PGF2α injection at the time of CIDR removal on the reproductive performance of Ghezel ewes. Estrus synchronization was performed using an intravaginal application of CIDR for 14 days and injection of 500 IU of PMSG at the time of CIDR removal. A total of 114 healthy fat-tailed ewes were randomly allotted into three groups as follow: control group (n = 35), did not receive any additional treatment; PG group (n = 44), each ewe received a dose of PGF2α at the time of CIDR removal; and PG+GnRH group (n = 35), the ewes received a dose of PGF2α at the time of CIDR removal and a single dose of GnRH, five days post-mating (post-conceptional day (PCD)- 5). Body condition score (BCS) of total ewes was determined at the time of CIDR insert. Blood samples were collected on PCD-19 for determining the serum progesterone levels. All the ewes were examined by transrectal ultrasonography 30-35 days after mating for pregnancy diagnosis. The serum values for P4 concentration were in control, PG and PG+GnRH groups 6.34 ± 1.17, 9.19 ± 2.55 and 10.57 ± 2.0 ng/mL respectively. The PG+GnRH treatment significantly increased the litter size compared to the control group (P = 0.04), but there were no significant differences in another reproductive indices between experimental groups. The multiple birth, twin, fecundity rates and litter size (P = 0.05, p = 0.03 and p = 0.003 respectively) were significantly higher in ewes with BCS > 2 compared to ewes with BCS ≤ 2. It is concluded that GnRH on PCD-5 treatment in addition to PGF2α injection at the time of CIDR removal could improve reproductive performance of Qezel ewes during non-breeding season.

Analysis of gene expression patterns by microarray hybridization in blood mononuclear cells of SLA-DRB1 defined Canadian Yorkshire pigs.

BACKGROUND: The Swine Leukocyte Antigen (SLA) system encodes molecules for self-nonself discrimination and is associated with immune responses and disease resistance. Three lines of pigs defined by their SLA-DRB1 alleles were developed at the University of Guelph for xenotransplantation and immune response studies. The aim of this project was to explore the potential association between defined SLA-DRB1 alleles and gene transcriptional patterns of other immune-related genes in blood mononuclear cells. FINDINGS: Three SLA-DRB1 alleles were characterized using a RT-PCR-based sequencing method. The loci represented included a new allele, DRB1*04ns01. Next, microarray heterologous (bovine-porcine) hybridization together with qPCR were used to explore differential gene expression between SLA-DRB1-defined groups. Microarray analysis showed significant (p < 0.01) differential expression for 5 genes, mostly related to inflammation. Genes varied according to the comparison analyzed. Further testing with qPCR revealed the same trend of differential expression for 4 of the genes, although statistical significance was reached for only one. CONCLUSION: A new SLA-DRB1 allele was characterized. A potential association was found between SLA-DRB1 alleles and inflammation-related genes. However, the influence of other genes cannot be ruled out. These preliminary findings agree with other studies linking MHC haplotypes and inflammation processes, including autoimmune disease. The study provides an initial view of the biological interactions between the SLA complex and other immune-related genes. Future studies will focus on characterization of SLA-haplotypes associated with these particular alleles and the dynamics of the immune response to antigenic challenges.