FSH plays a critical role in granulosa cell (GC) proliferation and steroidogenesis through modulation by factors including bone morphogenetic proteins family, which belongs to transforming growth factor b (TGFB) superfamily. TGFBs are the key factors in maintaining cell growth and differentiation in ovaries. However, the interaction of FSH and TGFB on the GCs' proliferation and steroidogenesis remains to be elucidated. In this study, we have investigated the role of SMAD4, a core molecule mediating the intracellular TGFB/SMAD signal transduction pathway, in FSH-mediated proliferation and steroidogenesis of porcine GCs. In this study, SMAD4 was knocked down using interference RNA in porcine GCs. Our results showed that SMAD4-siRNA causes specific inhibition of SMAD4 mRNA and protein expression after transfection. Knockdown of SMAD4 significantly inhibited FSH-induced porcine GC proliferation and estradiol production and changed the expression of cyclin D2, CDK2, CDK4, CYP19a1, and CYP11a1. Thus, these observations establish an important role of SMAD4 in the regulation of the response of porcine GCs to FSH.
BackgroundA number of studies have evaluated the efficacy of deferred stenting vs immediate stenting in patients with ST‐segment elevation myocardial infarction, but the findings were not consistent across these studies. This meta‐analysis aims to assess optimal treatment strategies in patient with ST‐segment elevation myocardial infarction.Methods and ResultsWe searched the PubMed, EMBASE, and the Cochrane Library for studies that assessed deferred vs immediate stenting in patients with ST‐segment elevation myocardial infarction. Nine studies including 1456 patients in randomized controlled trials and 719 patients in observational studies were included in the meta‐analysis. No significant differences were observed in the incidence of no‐ or slow‐reflow between deferred stenting and immediate stenting in randomized controlled trials (odds ratio [OR] 0.51, 95%CI 0.17‐1.53, P=0.23, I2=70%) but not in observational studies (OR 0.13, 95%CI 0.06‐0.31, P<0.0001, I2=0%). Deferred stenting was associated with an increase in long‐term left ventricular ejection fraction (weighted mean difference 1.90%, 95%CI 0.77‐3.03, P=0.001, I2=0%). No significant differences were observed in the rates of major adverse cardiovascular events (OR 0.53, 95%CI 0.27‐1.01, P=0.06 [randomized OR 0.98, 95%CI 0.73‐1.30, P=0.87, I2=0%; nonrandomized OR 0.30, 95%CI 0.15‐0.58, P=0.0004, I2=0%]), major bleeding (OR=0.1.61, 95%CI 0.70‐3.69, P=0.26, I2=0%), death (OR=0.78, 95%CI 0.53‐1.15, P=0.22, I2=0%), MI (OR=0.97, 95%CI 0.34‐2.78, P=0.96, I2=35%) and target vessel revascularization (OR 0.97, 95%CI 0.40‐2.37, P=0.95, I2=24%), between deferred and immediate stenting.ConclusionsCompared with immediate stenting, a deferred‐stenting strategy did not reduce the occurrence of no‐ or slow‐reflow, death, myocardial infarction, or repeat revascularization compared with immediate stenting in patients with ST‐segment elevation myocardial infarction, but showed an improved left ventricular function in the long term.
G protein-coupled receptor 3 (Gpr3) is a member of G protein-coupled receptor rhodopsin family, which is present throughout the follicle within the ovary and functions as a critical factor for the maintenance of meiotic prophase arrest in oocytes by a Gs protein-mediated pathway. In the current paper, attempts were made to clone and characterize a gene encoding Gpr3 from pigs and investigate its expression pattern in tissues and the whole cumulus-oocyte complexes (COCs) in vitro maturation (IVM). Rapid amplification of cDNA ends and RT-PCR gave rise to the full sequence of Gpr3 gene with its length being 2101 bp nucleotides, including an open reading frame of 993 bp, encoding a 331 amino acid polypeptide with the molecular weight of 35.2 kDa. Homology search and sequence multi-alignment demonstrated that the putative porcine Gpr3 protein sequence shared a high identity with other animal Gpr3 orthologs, including several highly conservative motifs and amino acids. Real-time PCR analysis showed that the Gpr3 gene was expressed in tissues of cerebrum, cerebellum, hypothalamus, pituitary, ovary, oviduct, uterus, heart, liver, spleen, lung, kidney, muscle, fat, testis, thymus and granulosa cell, oocyte and COCs at different expression levels. The expression levels of this gene in oocyte, uterus, liver, fat, pituitary and brain were higher than that in other tissues. Interestingly, the mRNA and protein levels of Gpr3 in the whole COCs were down-regulated, and its mRNA expression levels were significantly and negatively correlated with the degrees of cumulus expansion (r = -0.937, P < 0.01) during IVM, suggesting its important roles in cumulus expansion and oocyte maturation.
ObjectiveAstragalus polysaccharides (APS) are active constituents of Astragalus membranaceus. In this study, we aimed to investigate the effects of APS on memory impairment in a diabetic rat model and their mechanisms.MethodsA diabetic model was established in 50 male Wistar rats with streptozotocin intra-peritoneal injection. A blood glucose level higher than 16.7 mmol/L obtained 72 hours after the injection was regarded as a successful diabetic model. The modeled rats were divided into model group, high, medium, and low doses of APS, and piracetam groups (positive control). A group of ten rats without streptozotocin-induced diabetes were used as a normal control. After respective consecutive 8-week treatments, the levels of blood fasting plasma glucose, insulin, hemoglobin A1c, memory performance, hippocampal malondialdehyde, and superoxide dismutase were determined.ResultsAfter the 8-week APS treatment, serum fasting plasma glucose, hemoglobin A1c, and insulin levels were decreased compared with those of the model group (P<0.05). Importantly, memory impairment in the diabetic model was reversed by APS treatments. In addition, hippocampal malondialdehyde concentration was lowered, whereas that of superoxide dismutase was higher after APS treatments.ConclusionAPS are important active components responsible for memory improvement in rats with streptozotocin-induced diabetes. The potential mechanism of action is associated with the effects of APS on glucose and lipid metabolism, and antioxidative and insulin resistance. APS are constituents of A. membranaceus that are potential candidate therapeutic agents for the treatment of memory deficit in diabetes.
ABSTRACT. Bone morphogenetic proteins (BMPs) are the key factors in maintaining cell growth and differentiation in ovaries. BMPs initiate signaling by assembling BMP receptors and activating Smads, which in turn alter the expression of target genes. However, little is known about the effect of the deletion of the Bone morphogenetic protein receptor type IB (BMPRIB) on porcine granulosa cell (GCs). The objective of this study was to determine the effects of BMPRIB gene silencing, by small interfering RNA (siRNA), on the apoptosis and steroidogenesis of porcine GCs, and the expression of cell cycle-related and apoptosis-related genes. Results indicate that the BMPRIB siRNA caused specific inhibition of BMPRIB mRNA expression after transfection. Knockdown of the BMPRIB gene significantly inhibited porcine GCs proliferation and estradiol production, while inducing apoptosis of porcine GCs. Additionally, the declined expression of the BMPRIB gene changed the expressions of CylinD2, Cdk2, Bcl-2, and Cyp19a1. These
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