This study was undertaken to evaluate the relationship between in vitro maturation and plasminogen activators (PAs) activity on porcine cumulus-oocytes complexes (COCs) exposed to oxidative stress. When COCs were cultured in maturation medium with hydrogen peroxide (H2O2), the proportion of the germinal vesicle breakdown (GVBD) and oocytes maturation were decreased with addition of H2O2, and were significantly (p<0.05) lower in medium with 0.1 mM H2O2 than control group. Also, the rate of degenerated oocytes was increased in as H2O2 concentration increased. When COCs were cultured for 48 h, three plasminogen-dependent lytic bands were observed: tissue-type PA (tPA); urokinase-type PA (uPA); and tPA-PA inhibitor (tPA-PAI). PA activity was quantified using SDS-PAGE and zymography. When H2O2 concentration was increased, tPA and tPA-PAI activities also increased in porcine oocytes cultured for 48 h, but not uPA. In other experiment, embryos were divided into three groups and cultured in (1) control medium, (2) control medium with 1.0 mM H2O2 and (3) control medium with 1.0 mM H2O2 along with catalase in concentrations of 0.01, 0.1, and 1.0 mg/ml, respectively. H2O2 decreased the rate of GVBD and maturation in porcine COCs but catalase revealed protective activity against oxidative stress caused by H2O2. In this experiment, tPA and tPA-PAI activities were higher in media with 1.0 mM H2O2 alone. Increasing concentration of catalase decreased tPA and tPA-PAI activities in porcine oocytes. These results indicate that the exposure of porcine follicular oocytes to ROS inhibits oocytes maturation to metaphase-II stage and increase the oocytes degeneration. Also, we speculated that increased ROS level may trigger tPA and tPA-PAI activities in porcine oocytes matured in vitro.

Background : This study was performed to investigate the morphological and growth characteristics and metabolomics approach for the diploid and tetraploid Polygonum multiflorum Thunb (PMT). Methods and Results : The present study was performed to investigate the effective induction method of tetraploid plants using colchicine treatment. To know the morphological and growth characteristics of PMT overground growing was investigated about the stem diameter, leaf length and width, a scape, inflorescence length, chlorophyll, seed length and seed width. Metabolite profiles from P. multiflorum were analyzed by an ultra performance liquid chromatography–quadrupole - time of flight (UPLC-Q-TOF) mass spectrometry (MS). The number of stomata and chlorophyll content was also higher in tetraploid. However, the physiological and morphological characteristics of the newly developed tetraploid showed noticeable results compared to the diploid plant. Also, the inherent characteristics of a newly acquired tetraploid also exhibited better results compared to diploid. Twenty one compounds including catechins from leaf and thirty compounds including stilbenes from roots were identified as major metabolites of PMT. The levels of most root metabolites except dethiobiotin, epigallocatechin gallate, procyanidin B1, epiafzelechin 3-O-gallate, and moupinamide from PMT were higher than those of diploid. The levels of 2,3-digalloylglucose, fallacinol, tricin, physcion-8-O-β-D-glucoside, torachrysone-8-O-β-D-glucopyranoside, questin, aloe-emodin 8-O-(6′-O-acetyl)-glucoside, polydatin, 2,5-dimethyl-7-hydroxychromone, 3,5,4'-trihydroxystilbene, aloe-emodin, myrciacitrin II and ω-hydroxyemodin from teraploid PMT were 342, 32, 31, 21, 11, 11, 9, 8, 6, 5, 4, and 2 times, respectively, higher. Conclusion : From above results, overground growing of tetraploid P. multiflorum is fairly better than that of native P. multiflorum.