Eryngial (trans-2-dodecenal), a bioactive compound from Eryngium foetidum: its identification, chemical isolation, characterization and comparison with ivermectin in vitro

Parasitology ◽  
2013 ◽  
Vol 141 (2) ◽  
pp. 269-278 ◽  
Author(s):  
W. M. FORBES ◽  
W. A. GALLIMORE ◽  
A. MANSINGH ◽  
P. B. REESE ◽  
R. D. ROBINSON
Keyword(s):  
Larval Mortality ◽  
Anthelmintic Activity ◽  
Bioactive Compound ◽  
Strongyloides Stercoralis ◽  
Probit Analysis ◽  
The Other ◽  
Significant Difference ◽  
Third Stage ◽  
Eryngium Foetidum

SUMMARYMethanol-water (4:1, v/v) crude extracts (50 mg mL−1) of 25 Jamaican medicinal plants were screened in vitro for anthelmintic activity using infective third-stage larvae of Strongyloides stercoralis. The most effective extract was further chemically scrutinized to isolate and identify the source of the bioactivity, and the efficacy of this compound was compared with ivermectin. Eosin exclusion (0·1 mg mL−1) served as the indicator of mortality in all bioassays. A crude extract of Eryngium foetidum (Apiaceae) was significantly (Probit Analysis, P<0·05) more potent than the other plant extracts, taking 18·9 h to kill 50% (LT50) of the larvae. Further, the petrol extract of E. foetidum was significantly more effective (Probit Analysis, P<0·05) at killing the larvae (LT50, 4·7 h) than either its methanol–water or dichloromethane extract. The latter two effected less than 1% larval mortality after 120 h. With bioassay-driven column chromatography of the petrol extract, trans-2-dodecenal (eryngial) was identified and chemically isolated as the main anthelmintic compound in E. foetidum. There was a significant difference between the 24 h LD50 values (mm) of trans-2-dodecenal (0·461) and ivermectin (2·251) but there was none between the 48 h LD50 values (mm): trans-2-dodecenal (0·411) and ivermectin (0·499) in vitro.

In vitro nematicidal effects of medicinal plants from the Sierra de Huautla, Biosphere Reserve, Morelos, Mexico against Haemonchus contortus infective larvae

2008 ◽  
Vol 82 (1) ◽  
pp. 25-31 ◽  
Author(s):  
U. López-Aroche ◽  
D.O. Salinas-Sánchez ◽  
P. Mendoza de Gives ◽  
M.E. López-Arellano ◽  
E. Liébano-Hernández ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Larval Mortality ◽  
Evaluation Criteria ◽  
Anthelmintic Activity ◽  
Biosphere Reserve ◽  
The Other ◽  
Post Exposure ◽  
Infective Larvae ◽  
Third Stage

AbstractTwenty extracts from plants from Sierra de Huautla Biosphere Reserve, Morelos, Mexico were evaluated against Haemonchus contortus infective larvae in an in vitro assay. The plant species evaluated were Bursera copallifera, B. grandifolia, Lippia graveolens, Passiflora mexicana, Prosopis laevigata, Randia echinocarpa and Urtica dioica. The plants were separated into their parts and macerated with different solvents (n-hexane, acetone, ethanol and methanol). An in vitro assay was used to evaluate the anthelmintic activity against unsheathed third stage H. contortus infective larvae. The experiment was carried out in 24-well cell culture plates at room temperature with three replicates per treatment and using a concentration of 20 mg ml− 1. Ten 5 μl aliquots were taken from the corresponding wells and deposited on a slide for microscopical observation at 24, 48, 72 and 96 h post-exposure. The evaluation criteria were based on the average numbers of live and/or dead larvae in the different treatments. Alive and dead larval numbers were statistically analysed through the ANOVA test (P>0.01). The Tukey test was used as a complementary tool to determine which treatment was different from the other treatments (P>0.05). The highest mortality was observed with P. laevigata hexanic extract from stem and leaves combined, which produced 51%, 81% and 86% larval mortality at 24, 48 and 72 h post-exposure, respectively. On the other hand, B. copallifera stem acetonic extract exhibited 18%, 59% and 66% nematicidal activity after 24, 48 and 72 h of exposure, respectively.

scholarly journals In vitro Anthelmintic Activity of Two Aloe-derived Active Principles against Sheep Gastrointestinal Nematodes

2017 ◽  
Vol 12 (12) ◽  
pp. 1934578X1701201
Author(s):  
Gianluca Fichi ◽  
Matteo Mattellini ◽  
Elisa Meloni ◽  
Guido Flamini ◽  
Stefania Perrucci
Keyword(s):  
Larval Development ◽  
Larval Mortality ◽  
Plant Secondary Metabolites ◽  
Anthelmintic Activity ◽  
Gastrointestinal Nematodes ◽  
Egg Hatch ◽  
Reference Drug ◽  
Faecal Samples ◽  
Aloe Emodin

The in vitro anthelmintic activity on sheep gastrointestinal strongyle (GIS) eggs and larvae of 0.5% aloin and 0.1% aloe-emodin was investigated. From fresh faecal samples collected by ewes naturally infected by Haemonchus, Trichostrongylus and Teladorsagia nematodes, GIS eggs were isolated and cultivated in Petri dishes (100 eggs/dish). For the in vitro evaluation of the anthelmintic activity of tested compounds, the Egg hatch test (EHT), the Larval development test (LDT) and the Larval mortality/paralysis test (LMT) were used. In each assay, the activity of tested compounds was compared to untreated and treated (0.1% thiabendazole, TBZ) controls. Six repetitions were made through the experiment. Obtained data were statistically elaborated using the X2 test. In EHT, 0.5% aloin gave highly significantly different (P<0.01) results from the untreated controls. In LDT, both 0.1% aloe-emodin and 0.5% aloin almost completely prevented the larval development from L1 to L3, showing no significant differences (P<0.01) when compared to TBZ. In LMT, larval mortality observed in 0.5% aloin treated plates was significantly higher (P<0.01) than that observed in TBZ treated controls. These results show the in vitro anthelmintic properties on sheep GIS of the examined plant secondary metabolites. In LDT and/or LMT, the activity of 0.5% aloin and 0.1% aloe-emodin was comparable to or higher than that of the reference drug.

scholarly journals Natural Compounds from the Marine Brown Alga Caulocystis cephalornithos with Potent In Vitro-Activity against the Parasitic Nematode Haemonchus contortus

Pathogens ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 550
Author(s):  
Aya C. Taki ◽  
Robert Brkljača ◽  
Tao Wang ◽  
Anson V. Koehler ◽  
Guangxu Ma ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Brown Alga ◽  
Parasitic Nematode ◽  
Anthelmintic Activity ◽  
Adult Males ◽  
Terrestrial Plants ◽  
Third Stage ◽  
Future Work ◽  
Marine Brown Alga

Eight secondary metabolites (1 to 8) were isolated from a marine sponge, a marine alga and three terrestrial plants collected in Australia and subsequently chemically characterised. Here, these natural product-derived compounds were screened for in vitro-anthelmintic activity against the larvae and adult stages of Haemonchus contortus (barber’s pole worm)—a highly pathogenic parasitic nematode of ruminants. Using an optimised, whole-organism screening system, compounds were tested on exsheathed third-stage larvae (xL3s) and fourth-stage larvae (L4s). Anthelmintic activity was initially evaluated on these stages based on the inhibition of motility, development and/or changes in morphology (phenotype). We identified two compounds, 6-undecylsalicylic acid (3) and 6-tridecylsalicylic acid (4) isolated from the marine brown alga, Caulocystis cephalornithos, with inhibitory effects on xL3 and L4 motility and larval development, and the induction of a “skinny-straight” phenotype. Subsequent testing showed that these two compounds had an acute nematocidal effect (within 1–12 h) on adult males and females of H. contortus. Ultrastructural analysis of adult worms treated with compound 4 revealed significant damage to subcuticular musculature and associated tissues and cellular organelles including mitochondria. In conclusion, the present study has discovered two algal compounds possessing acute anthelmintic effects and with potential for hit-to-lead progression. Future work should focus on undertaking a structure-activity relationship study and on elucidating the mode(s) of action of optimised compounds.

scholarly journals Isorhamnetin: A Nematocidal Flavonoid from Prosopis laevigata Leaves Against Haemonchus contortus Eggs and Larvae

Biomolecules ◽  
2020 ◽  
Vol 10 (5) ◽  
pp. 773
Author(s):  
Edgar Jesús Delgado-Núñez ◽  
Alejandro Zamilpa ◽  
Manasés González-Cortazar ◽  
Agustín Olmedo-Juárez ◽  
Alexandre Cardoso-Taketa ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Laser Scanning ◽  
Larval Mortality ◽  
Small Ruminants ◽  
Environmental Scanning Electron Microscopy ◽  
Anthelmintic Activity ◽  
Ethyl Acetate Fraction ◽  
Confocal Laser ◽  
Eggs And Larvae

Haemonchus contortus affect small ruminants all over the world. Anthelmintics cause resistance, contamination, and a risk of public health. Prosopis laevigata is a plant used as a home remedy against many diseases in Mexico. This study arose from a preliminary study where a P. laevigata hydroalcoholic extract (Pl-hae) showed anthelmintic activity (aa) against H. contortus. Searching for bioactive compounds (bac) with high aa, the Pl-hae was fractioned obtaining an aqueous (Aq-F) and an ethyl acetate fraction (EtAc-F), and a flavonoid with aa identified as isorhamnetin was obtained from EtAc-F. Both fractions were in vitro assessed by the egg hatch test (eht) and larval mortality (lm) assays. The bac obtained from EtAc-F were characterised by NMR analysis. The highest aa were recorded with EtAc-F, resulting in 100% eht and 80.45% lm at 0.75 and 30 mg/mL, respectively. Alterations in eggs and larvae attributed to isorhamnetin were recorded by environmental scanning electron microscopy, confocal laser scanning and by high-resolution digital-coupled camera. This flavonoid caused 100% eht at 0.07 mg/mL after 48 h and 100% lm at 7.5 mg/mL after 72 h exposure. Isorhamnetin has promising potential as an anthelmintic against sheep haemonchosis.

Effect of Different Intermediary Bases on Microleakage of a Restorative Material in Class II Box Cavities of Primary Teeth

2017 ◽  
Vol 40 (2) ◽  
pp. 82-87 ◽  
Author(s):  
Faika Y. Abdelmegid ◽  
Fouad S. Salama ◽  
Waleed M. Al-Mutairi ◽  
Saud K. Al-Mutairi ◽  
Sultan O. Baghazal
Keyword(s):  
Primary Teeth ◽  
In Vitro Study ◽  
Class Ii ◽  
Positive Control ◽  
Negative Control ◽  
The Other ◽  
Control Groups ◽  
Significant Difference ◽  
The Mean

Introduction The aim of this in vitro study was to assess and compare the effect of different intermediary bases on microleakage between tooth and a nanocomposite interface in Class II box cavities in primary teeth. Methods Standard Class II box cavities were prepared in 52 primary molars and randomly divided into 9 groups according to the intermediary base used (Multicore Flow, Fuji II LC, SDR, Smart Dentin Replacement, and Biodentine). All specimens were subjected to thermocycling and prepared for microleakage testing and evaluation. Results There was significant difference in the mean ranks of microleakage between the 9 groups, which was observed in the gingival side (p<0.0001) and the occlusal side (p<0.0001). The mean ranks microleakage was significantly higher with experimental SDR, experimental Multicore Flow, and positive control materials when compared with the other 6 groups. The microleakage mean ranks were statistically significantly lower in experimental Fuji II LC, experimental Biodentine, and all negative control groups when compared with the other 3 groups. Conclusions Microleakage is affected by the application of intermediate material. Experimental Biodentine and Fuji II LC showed the lowest microleakage while experimental SDR and experimental Multicore Flow showed the highest microleakage.

Angiopoietin-1 and Osteopontin Expression by CD138+ Myeloma Cells Rather Than VEGF and CD45 Correlates with Bone Marrow Angiogenesis in Multiple Myeloma Patients at the Diagnosis.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 2501-2501
Author(s):  
Nicola Giuliani ◽  
Simona Colla ◽  
Francesca Morandi ◽  
Sabrina Bonomini ◽  
Mirca Lazzaretti ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
Linear Trend ◽  
The Other ◽  
Vegf Mrna ◽  
Myeloma Cells ◽  
Other Hand ◽  
Significant Difference ◽  
Angiopoietin 1

Abstract Bone marrow (BM) angiogenesis is increased in Multiple Myeloma (MM) patients and correlates with disease progression and patient survival. Myeloma cells secrete the main endothelial growth factor VEGF. In mouse models VEGF secretion as well as the angiogenic properties of MM cells correlate with the lack of CD45 expression by MM cells. However, recent data indicate that VEGF plasma cell expression is similar between MGUS and MM patients suggesting that other molecules could be involved. In line with this hypothesis we have recently demonstrated that myeloma cells may also produce factors with angiogenic properties as angiopoietin-1 (ANG-1) and osteopontin (OPN) that are involved in myeloma induced angiogenesis in vitro. In order to identify which factors correlate with BM angiogenesis in MM patients, we have investigated in a cohort of 121 newly diagnosed MM patients (stage I–III) the expression of the angiogenic molecules VEGF, ANG-1 and OPN and their correlation with bone marrow (BM) angiogenesis and CD45 expression by MM cells. We found that 90% of CD138+ MM cells tested were positive for VEGF mRNA. On the other hand we found that 50% and 40 % of MM patients were positive for ANG-1 and OPN mRNA respectively. Using the previously published cut off for CD45 expression we found that 61 out of 121 MM patients were positive for CD45 and 60 out of 121 were negative for CD45 expression. Any correlation was not observed between VEGF expression and BM angiogenesis in MM patients (p=0.5), whereas the number of microvessels X field was higher in Ang-1 positive patients in comparison with Ang-1 negative ones (mean±SE: 6.23±0.2 vs. 2.94±0.1, median: 6.21 vs. 2.79; p=0.001,) and the microvascular density (MVD) was significantly increased (32.98±1.7 vs. 14.55±1.3, median: 34.69 vs. 13.04; p&lt;0.01; capillaries: 26.73±1.3 vs. 10.42±0.8, median: 24.06 vs. 9.04; p&lt;0.01, small venules: 9.56 ±0.5 vs. 4.14±0.5, median: 10.60 vs. 3.65; p&lt;0.01). Furthermore a significantly positive correlation between Ang-1 expression and MVD was found (Pearson Chi-square: p=0.036, Cochran’s Linear Trend: p=0.01). A significantly higher MVD was also observed in the group of patients positive for OPN, (mean±SE: 29.1±0.7 vs. 17.55±0.37; p&lt;0.01) and similarly, the number of microvessels per field was higher in OPN positive patients in comparison with OPN negative ones (mean±SE: 6.7±0.15 vs. 4.28±0.04; p=0.05). On the other hand, any significant difference was not observed between CD45 positive and CD45 negative patients for the expression of VEGF (p=0.4), ANG-1 (p=0.3) and OPN (p=0.09). Consistently we did not find any significant difference in both MVD and number of vessels X field between CD45 positive patients as compared with CD45 negative ones (p=0.5 and p=0.4, respectively). Finally, a multivariate analysis confirmed that VEGF and CD45 did not correlate with the BM angiogenesis showing that ANG-1 expression by MM cells was more tightly correlated with MVD and the number of vessels X field as compared to OPN. Our data indicate that ANG-1 and in part OPN rather than VEGF and CD45 expression by MM cells are the critical determinants correlated with the increase of BM angiogenesis that occurs in MM patients at the diagnosis.

scholarly journals Anthelmintic Activity Ethanol Extract of Ocimum sanctum Linn. Leaves Against Ascaridia galli In Vitro

2019 ◽  
Vol 2 (1) ◽  
pp. 21
Author(s):  
Vanna Lidya Kharisma ◽  
Setiawan Koesdarto ◽  
Koesnoto Supriandono ◽  
Lucia Tri Suwanti ◽  
Sri Agus Sudjarwo ◽  
...  
Keyword(s):  
Exposure Time ◽  
Anthelmintic Activity ◽  
Ethanol Extract ◽  
Probit Analysis ◽  
Ocimum Sanctum ◽  
Ascaridia Galli ◽  
Time Interaction ◽  
Randomized Design ◽  
Lc50 And Lc90

The aims of this research are to determine concentration, exposure time, interaction between concentration and exposure time of ethanol extract of Ocimum sanctum Linn. Leaves to cause death toward Ascaridia galli in vitro, and the value of LC50 and LC90 ethanol extract of Ocimum sanctum Linn. Leaves. Research design that has been used in the research was completely randomized design. This research used 200 samples of Ascaridia galli with length 7-11 cm without differentiating their sex. The concentration ethanol extract of Ocimum sanctum Linn. leaves were 1.25%, 2.5%, 5%, 10%. The control was using CMC-Na 0.5%. Each treatment then being replicated four times. The observation and recording of dead worm were done at 0, 3, 6, 12 and 24 hours. Ascaridia galli were declared dead if there was no movement while disturbed by anatomy tweezers and dipped in slightly warm water (50ºC). The obtained data was analyzed using Anova Factorial and continued with Duncan Multiple Range Test by SPSS for Windows 22. The result were 10% concentration and exposure time for 24 hours caused the most mortality toward Ascaridia galli. Interaction between concentration and exposure time resulted 10% concentration ethanol extract of Ocimum sanctum Linn. leaves in 24 hours caused the most mortality towards Ascaridia galli. Probit analysis was used to calculate the LC50 and LC90 of Ocimum sanctum Linn. leaves. The results were LC50 ethanol extract of Ocimum sanctum Linn. leaves at 6 hours was 14.8%, at 12 hours was 4.8% and at 24 hours was 3.0% and the LC90 at 24 hours was 9.1%.

scholarly journals Cytotoxic Effect of Addition of Different Concentrations of Nanohydroxyapatite to Resin Modified and Conventional Glass Ionomer Cements on L929 Murine Fibroblasts

2021 ◽  
Author(s):  
Parisa Golkar ◽  
Ladan Ranjbar Omrani ◽  
Shide Zohourinia ◽  
Elham Ahmadi ◽  
Faeze Asadian
Keyword(s):  
Cytotoxic Effect ◽  
In Vitro Study ◽  
Biological Properties ◽  
The Other ◽  
Glass Ionomer Cements ◽  
Glass Ionomer ◽  
Beneficial Effects ◽  
Significant Difference ◽  
Murine Fibroblasts

Objectives: In this study we assessed the cytotoxic effect of nanohydroxyapatite (NHA) incorporated into resin modified and conventional glass ionomer cements (RMGICs and CGICs) on L929 murine fibroblasts. Materials and Methods: In this in vitro study, 0wt%, 1wt%, 2wt%, 5wt%, 7wt% and 10wt% concentrations of NHA were added to Fuji II LC RMGIC and Fuji IX CGIC powders. Eighteen samples (5×3mm) were fabricated from each type of glass ionomer, in six experimental groups (n=3): CG0, CG1, CG2, CG5, CG7, CG10, RMG0, RMG1, RMG2, RMG5, RMG7, and RMG10. Samples were incubated for 72h. The overlaying solution was removed and added to L929 fibroblasts. The methyl thiazolyl tetrazolium bromide (MTT) assay was performed at 24, 48 and 72h. The wavelength was read by a spectrophotometer. Data were analyzed by ANOVA and Tukey’s test. Results: There was no significant difference in cytotoxicity of the two types of glass ionomers, with and without NHA, except for CG0 and RMG0 groups after 72h. RMG0 group was significantly more cytotoxic than the CG0 group (P<0.05). In CG groups during the first 24h, the cytotoxicity of CG5 and CG7 groups was significantly higher than that of CG1; while, there was no significant difference between the RMG groups. Cytotoxicity significantly decreased in all groups after 24h (P<0.05). Conclusion: Incorporation of NHA into Fuji II LC RMGIC and Fuji IX CGIC did not affect their biocompatibility and therefore its addition to these materials can provide favorable biological properties, especially considering its beneficial effects on the other properties of GICs.

scholarly journals Anthelmintic Activity of Hymenodictyon pachyanta Stem Bark Extracts against Haemonchus Contortus

2019 ◽  
Vol 63 (2) ◽  
pp. 30-36
Author(s):  
D. O. Olayemi ◽  
M. M. Onakpa ◽  
O. C. Jegede
Keyword(s):  
Larval Development ◽  
Crude Extract ◽  
Haemonchus Contortus ◽  
Anthelmintic Activity ◽  
Stem Bark ◽  
Parasitic Nematodes ◽  
Egg Hatching ◽  
Crude Extracts ◽  
Significant Difference

Abstract The development of host resistance to anthelmintics and the increasing cost of commercial anthelmintics have encouraged the need for the in vitro anthelmintic evaluation of crude extract and fractions of Hymenodictyon pachyanta plant as alternative drugs against Haemonchus contortus. H. contortus is one of the most prevalent and highly pathogenic parasitic nematodes in small ruminant farming globally. H. pachyanta stem bark is a prospective plant used by the local and indigenous farmers of Nsukka, Enugu state, Nigeria. The stem bark of H. pachyanta were collected, dried, pulverized and extracted with 80 % methanol. The purpose of this study was to investigate the in vitro anthelmintic effects of these crude extract and fractions against H. contortus in sheep and goats. The two extracts (crude and fractions) of H. pachyanta were tested by the egg hatch assay (EHA) and the larval development inhibition assays (LDIA) and to compared the results with albendazole (as the positive control). The concentrations for the crude extract and albendazole used for this study were 0.78, 1.56, 3.125, 6.25 and 12.5 mg.ml−1. The results demonstrated that the crude extracts, fractions and albendazole all at the concentration doses of 12.5 mg.ml−1 produced 100 % inhibition of egg hatching and larval development. Statistically, there was no significant difference (P > 0.05) in the mean percentage inhibition of egg hatching and larval development inhibition of the crude extracts and fractions when compared with albendazole. However, a significant difference (P < 0.05) was observed with n-butanol fraction which inhibited 96.17 % of egg hatchability. All of the extracts and albendazole showed ovicidal and larvicidal effects and were able to induce over 50 % of the egg hatching and mortality of larvae at the concentration ranges of 0.78—12.5 mg.ml−1. The results obtained from our study suggest that H. pachyanta had ovicidal and larvicidal activity against H. contortus and that the bioactive plants compounds responsible for this effect could be attributed to the presence of tannins, alkaloids and the saponins contained in the crude extracts.

155 EFFECT OF CO-CULTURE WITH CUMULUS-DERIVED SOMATIC CELLS DURING IN VITRO MATURATION ON PORCINE CUMULUS–OOCYTE COMPLEXES AND SUBSEQUENT EMBRYONIC DEVELOPMENT AFTER IN VITRO FERTILIZATION

2014 ◽  
Vol 26 (1) ◽  
pp. 191 ◽  
Author(s):  
J. D. Yoon ◽  
L. Cai ◽  
S. U. Hwang ◽  
Y. Jeon ◽  
E. Kim ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Mrna Expression ◽  
In Vitro Maturation ◽  
Somatic Cells ◽  
The Other ◽  
Control Group ◽  
Cortical Granules ◽  
Developmental Potential ◽  
Significant Difference

The purpose of this study was to investigate the effects of co-culture with cumulus-derived somatic cells (CSC) during porcine in vitro maturation (IVM) and subsequent embryonic development after IVF. The CSC were cultured in Dulbecco's modified Eagle medium for 48 h with various numbers of cumulus-derived somatic cells (0.0, 2.5, 5.0, and 10.0 × 104), and then cultured in TCM-199 for 4 h before the oocytes were added. Cumulus-oocytes complexes from 3- to 6-mm follicles were matured in 500 μL of TCM-199, with eCG and hCG, for 22 h, and then cultured in M199 without hormones for 22 h. Each experiment consisted of at least 4 replicates. Statistical analyses were carried out using SPSS 17.0 software (SPSS Inc., Chicago, IL). Percentage data were compared by one-way ANOVA, followed by Duncan's multiple range test. Data were presented as means ± s.e.m. Differences were considered to be significant if the P-value was 0.05. After IVM, no significant difference (P < 0.05) was observed in nuclear maturation rate among the 0.0, 2.5, 5.0, and 10.0 × 104 groups (88.0 ± 2.37, 81.5 ± 2.17, 87.0 ± 1.98 and 86.0 ± 1.93%, respectively). The 2.5 × 104 group showed a significant (P < 0.05) increase in intracellular glutathione (GSH) levels compared with that of the other groups. Intracellular reactive oxygen species (ROS) levels of mature oocyte in all groups showed no significant differences. The developmental competence of matured oocytes in all groups was evaluated after IVF. The 2.5 and 5.0 × 104 groups showed significantly (P < 0.05) high cleavage rates (60.0 ± 4.7 and 64.52 ± 5.9%, respectively) compared with the 0 and 10.0 × 104 groups (43.15 ± 5.0 and 53.8 ± 5.0%, respectively). The 2.5 × 104 group showed a significantly (P < 0.05) higher BL formation rate (35.7 ± 2.9) than control group (21.0 ± 3.8%, respectively), and higher total cell number (127.25 ± 7.7) compared with the 0 and 10 × 104 groups (89.3 ± 4.0 and 92.6 ± 3.7, respectively). In the analysis of gene expression, IVF-BL derived from the 2.5 and 5.0 × 104 groups showed higher (P < 0.05) mRNA expression of PCNA, which is an essential component of the DNA replication and repair machinery and POU5F1 has been used to evaluate developmental potential in embryos. The 10.0 × 104 group showed higher (P < 0.05) mRNA expression of caspase-3 and Bak as known pro-apoptotic factors, compared with the control group IVF-BL. The results of cortical granules distribution which leads digesting sperm receptor proteins ZP2 and ZP3 to block polyspermy, showed that the 2.5 × 104 group was increased significantly (P < 0.05) compared with the other co-culture groups (13.7 ± 6.1, 29.2 ± 9.5, 18.3 ± 0.8 and 19.52 ± 5.3, respectively). In conclusion, co-culture with 2.5 × 104 cumulus-derived somatic cells during IVM improved the developmental potential of porcine IVF embryos by increasing the intracellular GSH level and distribution of cortical granules during oocyte maturation. This work was supported, in part, by a grant from the Next-Generation BioGreen 21 Program (No. PJ00956901), Rural Development Administration, and the National Research Foundation of Korea Grant funded by the Korean Government (NRF-2012R1A1A4A01004885, NRF-2013R1A2A2A04008751), Republic of Korea.

Sign in / Sign up

Export Citation Format

Share Document