Free Radical Scavenging Agents Research Articles

The positive effect of eugenol on acute pancreatic tissue injury: a rat experimental model.

Introductionwe present a rat experimental model used to evaluate the possible reduction in the extent of pancreatic tissue injury in acute pancreatitis cases, after administration of eugenol.Methodsone hundred and twenty Wistar rats were used, which were randomly assigned in 3 groups: sham (n=20), control (n=50) and eugenol (n=50). Acute pancreatitis was induced by biliopancreatic ligation in the control and eugenol groups, but not in the Sham group. In the eugenol group, eugenol was administered per-os. Five histopathological parameters, such as edema, inflammatory infiltration, duct dilatation, hemorrhage and acinar necrosis were evaluated.Resultsat 72 h from acute pancreatitis induction, the total histological score was diminished in the eugenol group (p<0.0005) and duct dilatation and inflammatory infiltration were reduced compared to the control group (p<0.05). In addition, at 72 h, eugenol reduced pancreatic myeloperoxidase activity (p<0.0005).Conclusioneugenol, a highly free radical scavenger agent, may have a preventive role in acute pancreatic injury, as it was evident in our rat experimental model.

The effect of Myrmecodia pendans extract toward gastric histology in white rats treated with toxic dose of gentamicin: a preliminary report

Introduction: Myrmecodia pendans, or traditionally known as ant nest fruit, is one of popular traditional medicines in Indonesia with a wide range of effects such as antidiabetic, anticancer, and free-radical scavenging agents. Despite of the extensive research, reports regarding its toxicity are scarce. Gentamicin is also one of popular and widely used anti-biotic but it is associated with a wide range of toxicity, especially if administered at high dose. Therefore, this study aimed to evaluate the effect of Myrmecodia pendans extract in the mice what received toxic dose of gentamicin.Method: 24 male white rats were used and randomly divided into 4 groups: P0 (negative control), P1 (100 mg/kgBW intraperitoneal gentamicin), P2 (gentamicyn + 250 mg/kgBW myrmercodia extract), P3 (250 mg/kgBW myrmercodia extract which administered for 7 days and followed by gentamicin). The treatment was given for 10 days and all mice were sacrificed at the end of the study. The stomach was taken from each mouse and examined pathologically using Hematoxylin-Eosin (HE) staining.Results: The necrosis was found in all groups that received gentamicin (P1-P3) with P2 had the most severe necrosis. Hemorrhage was only found in group that received Myrmecodia extract (P2 and P3) and, again, P2 had higher score of hemorrhage compared to P3.Conclusion: The administration of Myrmecodia extract at 250 mg/kgBW together with toxic dose of gentamicin exacerbate the gastric tissue damage in Wistar rats.

Green Tea Derivative EGCG Protects Against Radiation Induced Intestinal Injury By Modulating Nrf2 Signaling Pathway

Intestine is one of the most radio-sensitive tissues, and radiation induced intestinal injury (RIII) remains one of the most prevalent dose-limiting toxicities in radiotherapy for pelvic and abdominal cancers. The failure of synthetic compounds for efficient prevention or treatment of RIII led to growing interest in the study of natural derivatives from nontoxic plants. Epigallocatechin-3-gallate (EGCG), the major polyphenol in green tea, is a free radical scavenger and potent antioxidant agent, and has been widely studied in cancer chemoprevention. However, whether EGCG has the therapeutic effect on RIII remains unknown. Male C57BL/6J mice were treated with 9Gy of total body irradiation (TBI). 25mg/kg of EGCG were intraperitoneally injected before (once a day for 5 days) and 30minutes after TBI. Then, the survival rate and body weight were recorded. Meanwhile, jejunum tissues were collected and the severity of RIII was measured by HE staining, immunohistochemistry (IHC), immunofluorescence (IF) and TUNEL. Next, human intestinal epithelial cells (HIEC) were treated with EGCG and irradiated. Then, colony formation assay, flow cytometry, IF were conducted, and the level of reactive oxygen species (ROS) was assessed. Further, the expression of pivotal anti-oxidative response regulator nuclear factor erythroid 2 related factor 2 (Nrf2) was detected by IF and western blotting. Compared to the control, EGCG treatment significantly improved the survival time (from 6.1days to 10.4days, P<0.05) and increased the body weights of mice after TBI. HE staining of jejunum tissues showed that EGCG mitigated RIII, as reflected by the dramatic attenuation of crypt-villi architecture destruction. IHC results showed that EGCG treatment remarkably increased the Ki67-positive (from 15.7 to 30.1, P<0.001) and Lgr5-positive (from1.5 to 3.0, P<0.001) cells per crypt, indicating that EGCG promoted proliferation and survival of intestinal stem cells after TBI. Meanwhile, in EGCG treatment group the number of TUNEL-positive cells (from 4.9 to 1.0, P<0.001), γH2AX foci (from 2.8 to 0.4, P<0.001) and 8-OHdG-positive cells (from 3.5 to 0.6, P<0.001) significantly decreased per crypt, indicating that EGCG inhibited radiation-induced apoptosis and DNA damage in intestine. Consistently, the EGCG treatment remarkably improved the colony forming ability, while decreased apoptosis ratio, γH2AX foci and 8-OHdG foci of irradiated HIEC cells. For the mechanism study, EGCG treatment diminished the radiation-induced production of ROS, and promoted the nuclear translocation of key anti-oxidative response regulator Nrf2 of irradiated HIEC cells, indicating that EGCG scavenged radiation-induced ROS, and subsequently attenuated ROS-regulated intestinal oxidative DNA damage. Collectively, our findings suggested that EGCG protects against RIII by reducing ROS-induced DNA damage, and might be a novel therapeutic option for the clinical treatment of RIII.

Effects of N-Acetylcysteine and N-Acetylcysteine Amide on Erythrocyte Deformability and Oxidative Stress in a Rat Model of Lower Extremity Ischemia-Reperfusion Injury.

N-acetylcysteine (NAC) is an antioxidant which works as a free radical scavenger and antiapoptotic agent. N-acetylcysteine-amide (NACA) is a modified form of NAC containing an amide group instead of a carboxyl group of NAC. Our study aims to investigate the effectiveness of these two substances on erythrocyte deformability and oxidative stress in muscle tissue. Materials and Methods. A total of 24 Wistar albino rats were used in our study. The animals were randomly divided into five groups as control (n: 6), ischemia (n: 6), NAC (n: 6), and NACA (n: 6). In the ischemia, NAC, and NACA groups, 120 min of ischemia and 120 min of reperfusion were achieved by placing nontraumatic vascular clamps across the abdominal aorta. The NAC and NACA groups were administered an injection 30 min before ischemia (100 mg/kg NAC; 100 mg/kg NACA; intravenous). Blood samples were taken from the animals at the end of the ischemic period. The lower extremity gastrocnemius muscle was isolated and stored at −80 degrees to assess the total antioxidant status (TAS), total oxidant status (TOS), and oxidative stress index (OSI) values and was analyzed. Results. The erythrocyte deformability index was found to be statistically significantly lower in rats treated with NAC and NACA before ischemia-reperfusion compared to the groups that received only ischemia-reperfusion. In addition, no statistically significant difference was found between the control group and the NAC and NACA groups. The groups receiving NAC and NACA before ischemia exhibited higher total antioxidative status and lower total oxidative status while the oxidative stress index was also lower. Conclusion. The results of our study demonstrated the protective effects of NAC and NACA on erythrocyte deformability and oxidative damage in skeletal muscle in lower extremity ischemia-reperfusion. NAC and NACA exhibited similar protective effects on oxidative damage and erythrocyte deformability.

Protective Effect of Astaxanthin on Blue Light Light-Emitting Diode-Induced Retinal Cell Damage via Free Radical Scavenging and Activation of PI3K/Akt/Nrf2 Pathway in 661W Cell Model.

Light-emitting diodes (LEDs) are widely used and energy-efficient light sources in modern life that emit higher levels of short-wavelength blue light. Excessive blue light exposure may damage the photoreceptor cells in our eyes. Astaxanthin, a xanthophyll that is abundantly available in seafood, is a potent free radical scavenger and anti-inflammatory agent. We used a 661W photoreceptor cell line to investigate the protective effect of astaxanthin on blue light LED-induced retinal injury. The cells were treated with various concentrations of astaxanthin and then exposed to blue light LED. Our results showed that pretreatment with astaxanthin inhibited blue light LED-induced cell apoptosis and prevented cell death. Moreover, the protective effect was concentration dependent. Astaxanthin suppressed the production of reactive oxygen species and oxidative stress biomarkers and diminished mitochondrial damage induced by blue light exposure. Western blot analysis confirmed that astaxanthin activated the PI3K/Akt pathway, induced the nuclear translocation of Nrf2, and increased the expression of phase II antioxidant enzymes. The expression of antioxidant enzymes and the suppression of apoptosis-related proteins eventually protected the 661W cells against blue light LED-induced cell damage. Thus, our results demonstrated that astaxanthin exerted a dose-dependent protective effect on photoreceptor cells against damage mediated by blue light LED exposure.

Antioxidant and Cytotoxicity Profile of the Selected Alcoholic Beverages Widely Consumed in the Maharashtra State of India

The health concern of the alcoholic beverages has remained a key issue to be addressed to the consumers. The aim of the present study was to assess the antioxidant activity and cytotoxicity of the selected 22 bottled commercial alcoholic brands widely consumed in the Maharashtra state of India. The result of the present study shows that the brands of wines followed by whiskies and rums were more effective as free radical scavenging agents, containing considerable amount of polyphenols. Overall, the vodaka brands were observed to be more cytotoxic as compared to other brands. With few exceptions, there exists a positive correlation in the amount of polyphenols and free radical scavenging activity, while the former was negatively correlated with cytotoxicity.

The protective effect of alpha lipoic acid (ALA) on social interaction memory, but not passive avoidance in sleep-deprived rats.

Sleep is involved in maintaining energy, regulating heat, and recovering tissues. Furthermore, proper cognitive functions need sufficient sleep. Many studies have revealed the impairment effect of sleep deprivation (SD) on cognitive functions including learning and memory. Alpha lipoic acid (ALA) is a potent free radical scavenger, biological antioxidant, and neuroprotective agent. Furthermore, ALA improves learning and memory performance, decreases oxidative stress, and enhances antioxidant biomarkers. In this study, we aimed to investigate the effect of ALA on social interaction and passive avoidance memories in sleep-deprived rats. Total sleep deprivation (TSD) apparatus was used to induce SD (for 24h). Three-chamber paradigm test and shuttle box apparatus were used to evaluate social interaction and passive avoidance memory, respectively. Rats' locomotor apparatus was used to assess locomotion. ALA was administered intraperitoneally at doses of 17 and 35mg/kg for 3 consecutive days. The results showed SD impaired both types of memories. ALA at the dose of 35mg/kg restored social interaction memory in sleep-deprived rats; while, at the dose of 17mg/kg attenuated impairment effect of SD. Moreover, ALA at the dose of 35mg/kg impaired passive avoidance memory in sham-SD rats and at both doses did not rescue passive avoidance memory in sleep-deprived rats. In conclusion, ALA showed impairment effect on passive avoidance memory, while improved social interaction memory in sleep-deprived rats.

Ilex khasiana - The silent holly species of Aquifoliaceae on its pharmacognostical importance as a free radical scavenger and antibacterial agent

Ilex khasiana is a rare and endangered medicinal plant endemic to northeast India. Remaining largely unexplored,it is purported to have a range of medicinal values and may soon join the medicinal plant hall of fame.Extracts of the leaves were prepared using solvents of increasing polarity, namelypetroleum ether (IKP), chloroform (IKC) and methanol (IKM).The preliminary phytochemical screening indicated the presence of alkaloids, phytosterols, triterpenoids, saponins, reducing sugars, glycosides and carbohydrates. Free radical-scavenging activity was assessed using2,2-diphenyl-1-picrylhydrazyl(DPPH). IKC exhibited the highest activity with an IC50 of 17.22 ± 1.87, followed by IKM with an IC50 of 26.93 ± 5.14 and IKP with an IC50 of 37.16 ± 5.11. Butylated hydroxytoluene (BHT)was used as the standard DPPH scavenger and showed an IC50 of 8.31 ±0.72. IKM and IKC also showed positive antibacterial activity againstGram-negative bacteria such asEscherichia coli, Pseudomonas aeruginosa, andKlebsiella pneumoniae, and a Gram-positive bacteriaBacillus subtilis. IKP did not indicate any inhibition against the selected bacteria. Our findings substantiate the basis for further investigations on the medicinal potentials of the plant.&#x0D;

Activity Guided Fractionation of Anchomanes difformis (Blume) Engl. (ARACEAE) Stem Ethanol Extract: in Search of Free Radical Scavenging Agents

Activity Guided Fractionation of Anchomanes difformis (Blume) Engl. (ARACEAE) Stem Ethanol Extract: in Search of Free Radical Scavenging Agents

Quinoline Based Monocarbonyl Curcumin Analogs as Potential Antifungal and Antioxidant Agents: Synthesis, Bioevaluation and Molecular Docking Study.

In search for new fungicidal and free radical scavenging agents, we synthesized a focused library of 2-chloroquinoline based monocarbonyl analogs of curcumin (MACs). The synthesized MACs were evaluated for in vitro antifungal and antioxidant activity. The antifungal activity was evaluated against five different fungal strains such as Candida albicans, Fusarium oxysporum, Aspergillus flavus, Aspergillus niger, and Cryptococcus neoformans, respectively. Most of the synthesized MACs displayed promising antifungal activity compared to the standard drug Miconazole. Furthermore, molecular docking study on a crucial fungal enzyme sterol 14α-demethylase (CYP51) could provide insight into the plausible mechanism of antifungal activity. MACs were also screened for in vitro radical scavenging activity using butylated hydroxytoluene (BHT) as a standard. Almost all MACs exhibited better antioxidant activity compared to BHT.

Phytochemical and Pharmacological Profile of Terminalia chebula

Plant secondary metabolites have been of interest to man for a long time due to their pharmacological relevance. In this search Terminalia chebula is widely used to enhance the natural resistance to various diseases. It is called the “king of medicines” and always listed first in the Ayurvedic meteria medica because of its extraordinary powers of healing. Traditionally it has been used as a popular folk medicine for homeostatic, antitussive, laxative, diuretic and cardio tonic treatments. Recent studies showed that it has also been used as antidiabetic, antiviral, cardioprotective, anticancer, antioxidant, free radical scavenging and hypolipidemic agent. The present review is an attempt to highlight the various ethnopharmacological and traditional uses as well as phytochemical and pharmacological aspect of Terminalia chebula.

Enterochromaffin cells as the source of melatonin: key findings and functional relevance in mammals

The enteroendocrine cells in gastrointestinal (GI) tract synthesize more than thirty hormones in mammals. Among these cells, the enterochromaffin (EC) cells are probably the most important one due to the fact that they produce melatonin. The rate-limiting enzymes for melatonin synthesis including arylalkylamine-N-acetyltransferase (AANAT, currently the SNAT) and hydroxyindole-O-methyltransferase (HIOMT, currently the ASMT) have been identified in EC cells and this has confirmed the local melatonin production in GI tract by these cells. EC cells play a critical role in regulation of gastrointestinal physiology, particularly, in protection of the GI tract from free radical attack and inflammatory reaction. GI tract is the major site exposed to the oxidative stress and inflammation because of the food residue metabolism and the presence of trillions of microbes including the pathological bacteria. Thus, it requires strong protection. Melatonin synthesized by the EC cells provides the onsite protection in GI tract since this molecule is the potent free radical scavenger and effective ant-inflammatory agent. In this review we summarize the available information regarding the structural and functional variability of the EC cells as well as their pathophysiological roles in the GI tract. The focus is given to the protective effects of melatonin produced by the EC cells on the oxidative stress, inflammation and microbiota balance in GI tract.

In vitro and in vivo biocompatibility assessment of free radical scavenging nanocomposite scaffolds for bone tissue regeneration.

Bone is the second most transplanted tissue in the world, resulting in increased demand for bone grafts leading to the fabrication of synthetic scaffold grafting alternatives. Fracture sites are under increased oxidative stress after injuries, affecting osteoblast function and hindering fracture healing and remodeling. To counter oxidative stress, free radical scavenging agents, such as cerium oxide nanoparticles, have gained traction in tissue engineering. Toward the goal of developing a functional synthetic system for bone tissue engineering, we characterized the biocompatibility of a porous, bioactive, free radical scavenging nanocomposite scaffold composed of poly(1,8 octanediol-co-citrate), beta-tricalcium phosphate, and cerium oxide nanoparticles. We studied cellular and tissue compatibility utilizing in vitro and in vivo models to assess nanocomposite interactions with both human osteoblast cells and rat subcutaneous tissue. We found the scaffolds were biocompatible in both models and supported cell attachment, proliferation, mineralization, and infiltration. Using hydrogen peroxide, we simulated oxidative stress to study the protective properties of the nanocomposite scaffolds via a reduction in cytotoxicity and recovered mineralization of osteoblast cells in vitro. We also found after implantation in vivo the scaffolds exhibited biocompatible properties essential for successful scaffolds for bone tissue engineering. Cells were able to infiltrate through the scaffolds, the surrounding tissues elicited a minimal immune response, and there were signs of scaffold degradation after 30 days of implantation. After the array of biological characterization, we had confirmed the development of a nanocomposite scaffold system capable of supporting bone-remodeling processes while providing a protective free radical scavenging effect.

Synthesis, biological evaluation and molecular docking studies of novel 3,5-disubstituted 2,4-thiazolidinediones derivatives

A series of thirteen novel 2,4-thiazolidinedione derivatives were synthesized through three step reaction procedure. The title compounds were synthesized by Knoevenagel condensation at the 5th position of the 2,4-thiazolidinedione ring. Various physicochemical and spectral studies were conducted to characterize the synthesized derivatives including- IR, Mass, 1H NMR, 13C NMR and elemental analysis. The derivatives were screened for in vivo anti diabetic, in vivo anti-inflammatory and in vitro free radical scavenging activities by carrageenan induced rat paw edema method, alloxan induced diabetes in wistar rats method and FRAP (ferric reducing antioxidant power) method respectively. Some of the derivatives emerged out as potent antidiabetic, anti inflammatory and free radical scavenging agents. Molecular docking was carried out to investigate some possible structural insights into the potential binding patterns of the most potent anti-diabetic molecules NB7,NB12 and NB13 with the active sites of target PPARγ (PDB ID: 2PRG) using MOE software. Dichloro derivative compound NB-7 has shown great potential in the present study as it not only has maximum antidiabetic activity but also possess excellent anti-inflammatory and antioxidant potential.

PHYTOCHEMICAL AND BIOACTIVITY DIVERSITY IN THE EXTRACTS FROM BULBS AND LEAVES OF DIFFERENT POPULATIONS OF Allium jesdianum, A VALUABLE UNDERUTILIZED VEGETABLE

Allium jesdianum Boiss. &amp; Buhse (Yazdi onion) belonging the family Alliaceae, is an endemic species of Iran that grows wild in the Zagros Mountains range, southwestern Iran. The indigenous people of Iran use the leaves and bulbs of A. jesdianum for the treatment of colds and kidney problems. The bulbs and leaves of various populations of the plant were collected from the alpine regions in Chaharmahal va Bakhtiari province, southwestern Iran. The total phenolic content of ethanol extract was determined by Folin-Ciocalteu method, the antioxidant activity was evaluated measuring 1,1-diphenyl-2-picrylhydrazyl (DPPH), and the antibacterial activity of the extracts against four bacteria was determined by serial dilution assay. Results indicated that the total phenolic content in ethanol extracts from leaves and bulbs of A. jesdianum ranged between 27.83 to 98.23 mg GAE/g extract. A comparison of all plant extracts in the DPPH assay indicated that ethanol extracts from the populations of A. jesdianum leaves were the most effective free radical scavenging agents. The extracts indicated moderate-to-good inhibitory activities against four bacteria, especially against B. cereus. This finding suggests that the bulbs and leaves of A. jesdianum may be considered a natural source of antioxidants and antimicrobial agents.

Selenium⁻Fascinating Microelement, Properties and Sources in Food.

Selenium is a micronutrient that is essential for the proper functioning of all organisms. Studies on the functions of selenium are rapidly developing. This element is a cofactor of many enzymes, for example, glutathione peroxidase or thioredoxin reductase. Insufficient supplementation of this element results in the increased risk of developing many chronic degenerative diseases. Selenium is important for the protection against oxidative stress, demonstrating the highest activity as a free radical scavenger and anti-cancer agent. In food, it is present in organic forms, as exemplified by selenomethionine and selenocysteine. In dietary supplementation, the inorganic forms of selenium (selenite and selenate) are used. Organic compounds are more easily absorbed by human organisms in comparison with inorganic compounds. Currently, selenium is considered an essential trace element of fundamental importance for human health. Extreme selenium deficiencies are widespread among people all over the world. Therefore, it is essential to supplement the deficiency of this micronutrient with selenium-enriched food or yeast cell biomass in the diet.

Melatonin attenuates the detrimental effects of UVA irradiation in human dermal fibroblasts by suppressing oxidative damage and MAPK/AP-1 signal pathway in vitro.

People living in Mediterranean countries are mostly exposed to solar ultraviolet (UV) radiation that damages skin and results in photoaging which involves activation of epidermal growth factor receptor (EGFR) and downstream signal transduction through mitogen-activated protein kinases (MAPKs) in fibroblasts. Generation of reactive oxygen/nitrogen species by UV radiation is also critical for EGFR and MAPKs activation. MAPKs are responsible for activation of AP-1 subunits in the nucleus which induce matrix metalloproteinases. Melatonin, along with its metabolites, are known to be the most effective free radical scavenger and protective agent due to its ability to react with various radicals, lipophilic/hydrophilic structures. In this study, we investigated the effects of melatonin on UVA-irradiated primary human dermal fibroblasts (HDFs) by following the alteration of molecules from cell membrane to the nucleus and oxidative/nitrosative damage status of the cells in a time-dependent manner which have not been clearly elucidated yet. To mimic UVA dosage in Mediterranean countries, HDFs were exposed to UVA with sub-cytotoxic dosage (20J/cm2 ) after pretreatment with melatonin (1μmol/L) for 1hour. Changes in the activation of the molecules and oxidative/nitrosative stress damage were analyzed at different time points. Our results clearly show that melatonin decreases UVA-induced oxidative/nitrosative stress damage in HDFs. It also suppresses phosphorylation of EGFR, activation of MAPK/AP-1 signal transduction pathway and production of matrix metalloproteinases in a time-dependent manner. Melatonin can be used as a protective agent for skin damage against intracellular detrimental effects of relatively high dosage of UVA irradiation.

Melatonin restricts the viability and angiogenesis of vascular endothelial cells by suppressing HIF-1α/ROS/VEGF.

Angiogenesis is an essential process involved in various physiological, including placentation, and pathological, including cancer and endometriosis, processes. Melatonin (MLT), a well-known natural hormone secreted primarily in the pineal gland, is involved in regulating neoangiogenesis and inhibiting the development of a variety of cancer types, including lung and breast cancer. However, the specific mechanism of its anti-angiogenesis activity has not been systematically elucidated. In the present study, the effect of MLT on viability and angiogenesis of human umbilical vein endothelial cells (HUVECs), and the production of vascular endothelial growth factor (VEGF) and reactive oxygen species (ROS), under normoxia or hypoxia was analyzed using Cell Counting kit 8, tube formation, flow cytometry, ELISA and western blot assays. It was determined that the secretion of VEGF by HUVECs was significantly increased under hypoxia, while MLT selectively obstructed VEGF release as well as the production of ROS under hypoxia. Furthermore, MLT inhibited the viability of HUVECs in a dose-dependent manner and reversed the increase in cell viability and tube formation that was induced by hypoxia/VEGF/H2O2. Additionally, treatment with an inhibitor of hypoxia inducible factor (HIF)-1α (KC7F2) and MLT synergistically reduced the release of ROS and VEGF, and inhibited cell viability and tube formation of HUVECs. These observations demonstrate that MLT may serve dual roles in the inhibition of angiogenesis, as an antioxidant and a free radical scavenging agent. MLT suppresses the viability and angiogenesis of HUVECs through the downregulation of HIF-1α/ROS/VEGF. In summary, the present data indicate that MLT may be a potential anticancer agent in solid tumors with abundant blood vessels, particularly combined with KC7F2.

Effect of various culture conditions on shoot multiplication and GC–MS analysis of Plumbago zeylanica accessions for plumbagin production

Plumbago zeylanica, a pharmaceutically important medicinal plant, contains a wide range of phytocompounds. Culture parameters like carbon source, nitrogen source, and culture media are essential for the development and growth of explants. In this investigation, the influence of various carbon sources (sucrose, glucose, and fructose at 3% concentration), nitrogen source (ammonium nitrate, sodium nitrate, and potassium nitrate) and plant tissue culture media (MS medium, Gamborg’s B5 medium, White medium and Nitsch medium) on shoot multiplication of five different accessions was studied. Optimum growth of all five accessions was observed in MS media containing 3% sucrose and ammonium nitrate as a source of carbon and nitrogen. Out of five accessions, IC-524441 showed the highest shoot multiplication. Further, methanolic extracts of all accessions (grown in MS media containing 3% sucrose and ammonium nitrate as nitrogen source) were prepared and comparison of extracts in DPPH assay indicated that accession number IC-524441 was the most effective free radical scavenging agent. Total phenolic, flavonoid and tannin content ranges were from 20 to 70 µg/ml, 40 to 100 µg/ml and 55 to 120 µg/ml, respectively, and the highest amount was found in accession number IC-524441. Sucrose and ammonium nitrate content may be responsible for increased antioxidant activity, flavonoids content, phenolic content, and tannin content in accession number IC-524441. GC–MS of ethyl acetate extract of all five accessions of P. zeylanica was conducted (grown in MS media containing 3% sucrose and ammonium nitrate as nitrogen source). GC–MS analysis of the aerial part showed the presence of various phytocompounds, which include 1,4-naphthalenedione, 3-eicosene, 5-eicosene, phthalic acid, o-anisic acid, thioctic acid, 1-octadecene, 5-t-butyl-cycloheptene, 2-benzoyl-1,2-dihydro-1-isoquinolinecarbonitrile, octadecanal, silane, 3-methoxy-2-methyl-2-(1-phenyl-ethylamino)-propionic acid, and 1-nonadecene. Accession number IC-524441 contains the highest amount of plumbagin, i.e. 14.19 ± 0.5 µg/ml as compared to the others.

Melatonin supplementation during prolonged in vitro maturation improves the quality and development of poor-quality porcine oocytes via anti-oxidative and anti-apoptotic effects.

Poor-quality oocytes (those with 1-2 layers of cumulus cells) typically possess low meiotic competence and development. Prolonging the duration of in vitro maturation (IVM; 52 hr) can enhance the maturation rate of poor-quality oocytes, but it does not improve subsequent embryonic development. This likely reflects the increased reactive oxygen species (ROS) production and apoptosis seen in these oocytes compared with the non-prolonged IVM (44 hr) group. Melatonin is a free radical scavenger, anti-oxidant and anti-apoptotic agent that reported to enhance the quality of embryos by inhibiting ROS generation and apoptosis. Therefore, we herein investigated whether melatonin combined with prolonged IVM (52 hr) could improve the quality and development of poor-quality oocytes. We supplemented IVM and/or in vitro culture (IVC) media with various concentrations (0, 10-7 , 10-6 , 10-5 M) of melatonin, and estimated parameters related to oocyte quality and development. The addition of melatonin (10-6 M) to a prolonged IVM system improved the oocyte quality and development compared with those of the melatonin-free poor-quality oocytes group, and that this was due to decreases in ROS generation, apoptosis, and DNA damage. When melatonin was added during both IVM (10-6 M) and IVC (10-6 M), we observed a cumulative positive influence on the embryonic development and quality; this treatment enhanced the expression level of Oct4 and decreased the levels of ROS, DNA damage, and apoptosis. Together, these findings suggest that the combination of melatonin plus prolonged IVM can improve the quality and development of poor-quality porcine oocytes via anti-oxidative and anti-apoptotic effects.