ABSTRACT
Previous studies suggest that the inclusion of melatonin (MTn) in in vitro maturation protocols improves the developmental competence of oocytes by scavenging reactive oxygen species (ROS). However, the molecular mechanisms integrating melatonin receptor (MT)-mediated lipid metabolism and redox signaling during in vitro cumulus-oocyte complex (COC) development still remain unclear. Here, we aimed to elucidate the potential role of MTn receptors in lipid metabolic adjustments during in vitro porcine COC development. We observed that MTn-mediated Gsα-cAMP/PKA signaling facilitated lipolysis primarily through the MT2 receptor and subsequently increased fatty acid (FA) release by hydrolyzing intracellular triglycerides (TGs) in cumulus cells. Furthermore, CD36 was a critical FA transporter that transported available FAs from cumulus cells to oocytes and promoted de novo TG synthesis in the latter. In addition, MTn regulated lipogenesis and intracellular lipolysis to maintain lipid homeostasis and limit ROS production, thereby supporting oocyte cytoplasmic maturation and the subsequent embryo development. Taken together, these findings provide insight into the possible mechanism integrating MT2-mediated lipid homeostasis and redox signaling, which limits ROS production during in vitro COC development. Therefore, understanding the dynamics of the interactions between lipid homeostasis and redox signaling driven by MT2 is necessary in order to predict drug targets and the effects of therapeutics used to improve female reproductive health.
ABSTRACT
To investigate the effects of tannins (TA) on porcine oocyte in vitro maturation (IVM), different concentrations of TA (0, 1, 10 and 100 µg/mL) were supplemented with a maturation medium and the COCs and subsequent embryonic development were examined. The results showed that 10 µg/mL TA significantly improved the cumulus expansion index (CEI), cumulus-expansion-related genes (PTGS1, PTGS2, PTX-3, TNFAIP6 and HAS2) expression and blastocyst formation rates after parthenogenetic activation (PA), in vitro fertilization (IVF) and somatic cell nuclear transfer (SCNT) compared to the control groups, but not oocyte nuclear maturation. Nevertheless, 10 µg/mL TA dramatically enhanced the mRNA expression of oocyte-development-related genes (BMP15, GDF9, CDC2 and CYCLIN B1), GSH, ATP, SOD1, PGC1α, BMP15, GDF9 and CDC2 levels and reduced intracellular ROS level in porcine oocytes. These results indicated that porcine oocyte cytoplasmic maturation was improved by 10 µg/mL TA treatment during IVM. In contrast, a high concentration of TA (100 µg/mL) significantly decreased the CEI and PTGS1, PTGS2, PTX-3 and HAS2 mRNA expressions in cumulus cells, and reduced oocyte nuclear maturation and the total cell numbers/blastocyst. In general, these data showed that 10 µg/mL TA supplementation has beneficial effects on oocyte cytoplasmic maturation and subsequent embryonic development in pigs.
ABSTRACT
The aim of this study was to investigate the effect of the total saponins of Panaxginseng (TSPG) on cells expression of apoptosis-related genes bax and bcl-xl in HL-60 cells and its mechanism inducing apoptosis of HL-60 cells. The morphology of HL-60 cells was observed under normal and fluorescence microscopes; the percentage of apoptotic HL-60 cells was assayed by flow cytometry and the DNA ladder was observed by DNA agarose gel electrophoresis; the expression changes of bax and bcl-xl mRNAs were detected by RT-PCR after HL-60 cells were treated with TSPG at final concentrations of 0, 100, 200, 400, 800 and 1600 microg/ml for 48 hours. The results showed that the percentage of apoptotic HL-60 cells went up as the dose increased, the typical apoptotic cell morphology and the appearance of apoptotic DNA ladder could be observed when treated with 0 - 400 microg/ml TSPG for 48 hours. At the same time and same range of concentration, the expression of bax mRNA increased and the bcl-xl expression decreased gradually. When higher than 400 microg/ml of TSPG was used, cell necrosis appeared and the percentage of apoptotic HL-60 cells even decreased. It is concluded that the apoptosis or necrosis in HL-60 cells can be induced by TSPG at certain range of concentration, and the percentage of apoptosis is dose-dependent. The effect on up-regulation of bax mRNA and down-regulation of bcl-xl mRNA probably play an important role in apoptosis of HL-60 cells induced by TSPG.
Subject(s)
Humans , Apoptosis , HL-60 Cells , Panax , Chemistry , RNA, Messenger , Genetics , Metabolism , Saponins , Pharmacology , bcl-2-Associated X Protein , Genetics , MetabolismABSTRACT
13C metabolic flux analysis(13CMFA)have been the research hotspots of metabolic engineering internationally due to its accuracy and applicability.It is vital that the measurement of 13C labelling pattern of proteinogenic amino acids for 13C metabolic flux analysis.To acquire 13C-labelling proteinogenic amino acids,Pseudomonas denitrifican which products Vitamin B12 was firstly fed with mimimal culture medium contained 20% U-13C and 80% natural glucose,after the culture reached a steady state,then about 20 mg biomass was hydrolyzed by 1 ml of 6 mol/L hydrochloric acid for 24h at 110℃.Then amino acids was separated,concentrated,evaporated in a vacuum,and derivatized with MBDSTFA,TBDMS-derivatized amino acids can be observed by GC-MS last we get 13C labelling pattern of fifteen aminio acids through mass spectrum.The experimental methods and sample preparation offers referential value for the development of 13C metabolic flux analysis in our courtry.
ABSTRACT
Objective To increase the awareness of virus-associated hemophagocytic syndrome.Methods Sixteen cases of virus-associated hemophagocytic syndrome were retrospectively analyzed.Results All presented with persistent high fever,cytopenia,hepato-splenomegaly,hepatic dysfunction,hypertriglyceridemia,hyperferritinemia,coagulopathy,hypofibrinogenemia,cytokine storm and a low natural killer cell activity.All patients had lymphohistiocytic accumulation in bone marrow.Treatment with high-dose gamma-globulin and high-dose methylprednisolone.Clinical symptoms and laboratory improved,and five patients died.Conclusion Aggressive early diagnosis and treatment are critical to improve survival.
