Oil-in-water nanoemulsions loaded with lycopene extracts encapsulated by spray drying:Formulation,characterization and optimization

Lycopene is very susceptible to degradation once released from the protective chromoplast environment.In this study,oil-in-water(O/W)nanoemulsions coupled with spray drying technology were applied for the encapsulation and stabilization of lycopene extracted from tomato waste.Tomato extract was obtained by ultrasound-assisted extraction.Nanoemulsions were prepared by a high-speed rotor stator using isopropyl myristate as the oil phase and Pluronic F-127 as the emulsifier for the aqueous external phase.The effect of emulsification process parameters was investigated.Spray drying of the produced emulsions was attempted to obtain a stabilized dry powder after the addition of a coating agent.The effect of different coating agents(maltodextrin,inulin,gum arabic,pectin,whey and polyvinylpyrrolidone),drying temperature(120-170℃),and feed flow rate(3-9 ml·min^(-1))on the obtained particles was evaluated.Results revealed that the emulsion formulation of 20/80(O/W)with 1.5%(mass fraction)of Pluronic F-127 as stabilizer in the aqueous phase resulted in a stable nanoemulsion with droplet sizes in the range of 259-276 nm with a unimodal and sharp size distribution.The extract in the nanoemulsion was well protected at room temperature with a degradation rate of lycopene of about 50%during a month of storage time.The most stable emulsions were then processed by spray drying to obtain a dry powder.Spray drying was particularly successful when using maltodextrin as a coating agent,obtaining dried spherical particles with mean diameters of(4.87±0.17)μm with a smooth surface.The possibility of dissolving the spray dried powder in order to repristinate.The original emulsion was also successfully verified.

Molecular mechanism of lycopene cyclases regulating carotenoids ratio in different branches during tea leaf and flower development

Carotenoids are essential components in tea quality, contributing to leaf color and aroma. However, little information about carotenoids in different tea cultivars and their biosynthesis regulation mechanism during leaf development is known. Here we analyzed carotenoids by HPLC in the buds and leaves of 113 tea cultivars harvested on the same day. By profile clustering, carotenoids were divided into five groups. Same group cultivars displayed divergence in the total content of carotenoids but a similar molar ratio. To figure out the molecular mechanisms of this phenomenon, we further characterized all functional lycopene cyclases, which are the branch point of the carotenoid biosynthesis pathway. Two β-lycopene cyclases(CsLCYB1 and CsLCYB2) and one ε-lycopene cyclase(CsLCYE1) were cloned. Subcellular localization analysis showed that all cloned CsLCYs were localized in plastids. Enzyme activity assays in E. coli indicated both CsLCYBs catalyzed lycopene into β-carotene, and CsLCYE1 produced δ-carotene and ε-carotene. We found CsLCYB1 and CsLCYE1 predominantly expressed in leaf, while CsLCYB2was mainly expressed during flowering stages. Suppression by antisense oligonucleotides reduced CsLCYB1 and CsLCYE1 transcripts and led to reduction of both β,β-branch and β,ε-branch carotenoids in leaf. The expression levels of CsLCYB1 showed a significant positive correlation withβ,β-branch carotenoids in leaf. Our study provides carotenoid profiles of different tea cultivars, which can assist tea producers in selecting cultivars of interest. Meanwhile, we proposed the molecular mechanism of carotenoids reflecting the tenderness of tea plant leaf from a metabolic flux perspective, and suggested lycopene cyclase that could be applied to the breeding of tea varieties with different branch carotenoids.

Protective effect of lycopene on Parkinson's disease cell model based on endoplasmic reticulum stress

Objective:To evaluate the effect of lycopene on Parkinson's disease cell model and its possible mechanism.Methods:The SH-SY5Y cells were treated with 0.5μmol/L rotenone for 24 h to establish Parkinson's disease cell model.The experiments were randomly divided into the control group,the lycopene group,the rotenone group,the pretreatment groups of different concentrations lycopene(low,medium,high concentration).Cell viability was detected by CCK-8 assay,the morphological changes of cells were observed under an inverted microscope,Hoechst staining was used to observe cell apoptosis,the expression and distribution of endoplasmic reticulum stress marker proteins GRP78 and CHOP in each group were detected by Western blot and cell immunofluorescence.Results:The study found that compared with the control group,the cell viability in the rotenone group was significantly decreased with obvious apoptosis;compared with the rotenone group,the cell viability of the lycopene pretreatment group was improved,and the difference was statistically significant(P<0.05);The apoptosis in the lycopene pretreatment group was decreased.The expression of GRP78 and CHOP in the rotenone group was significantly higher than that in the control group(P<0.01),while the expression of both in the high concentration lycopene pretreatment group was lower than that in the rotenone group(P<0.05).Conclusion:Lycopene pretreatment had a significant protective effect on rotenone-induced SH-SY5Y cells,which may be related to the fact that lycopene pretreatment can effectively alleviate endoplasmic reticulum stress in SH-SY5Y cells damaged by rotenone.

RP-HPLC determination of lycopene in microcapsules

Aim A RP- HPLC method for determination of lycopene in microcapsules was established. Methods The HPLC assay was performed on an Alltima Cls （4.6 mm × 250 mm, 5μm） column with a mixture of methanol-THF-water （66：30：4, V/V/V） as mobile phase at a flow rate of 1.5 mL·min^-1 and the UV detection wavelength was 472 nm. Results The linear range of lycopene was 3.6-18 μg·mL^-1, r = 0.999 8, the average recovery was from 99.81% to 101.06% with RSD less than 1.83%. The RSD of intra-day and interday precision were less than 3.34%. Conclusion The method is simple, accurate and suitable for the determination of lycopene in microcapsules.

Determination of Lycopene Concentration in Dog Plasma by RP-HPLC

A method for determination of lycopene concentration in dog plasma wasestablished. Methods RP-HPLC was used; the mobile phase consisted of methanol-acetonitrile-methylenechloride (40:30:30, V/V) , the wavelength of detection was 472 nm, the column temperature wasambient temperature, and the flow rate was 1.0 mL·min^(-1). Results The standard curve was linearin the range from 0.012 4 to 0.496 μg·mL^(-1) with r=0.9992. The average extraction recovery was97.6% +-4.2%. The intra-day and inter-day RSD were 1.52% -4.95% and 2.31% -7.38%, respectively.Conclusion This method is sensitive, rapid, reproducible, and of good selectivity for the analysisof lycopene in dog plasma.

Possible mechanisms of lycopene amelioration of learning and memory impairment in rats with vascular dementia

Oxidative stress is involved in the pathogenesis of vascular dementia. Studies have shown that lycopene can significantly inhibit oxidative stress;therefore, we hypothesized that lycopene can reduce the level of oxidative stress in vascular dementia. A vascular dementia model was established by permanent bilateral ligation of common carotid arteries. The dosage groups were treated with lycopene(50, 100 and 200 mg/kg) every other day for 2 months. Rats without bilateral carotid artery ligation were prepared as a sham group. To test the ability of learning and memory, the Morris water maze was used to detect the average escape latency and the change of search strategy. Hematoxylin-eosin staining was used to observe changes of hippocampal neurons. The levels of oxidative stress factors, superoxide dismutase and malondialdehyde, were measured in the hippocampus by biochemical detection. The levels of reactive oxygen species in the hippocampus were observed by dihydroethidium staining. The distribution and expression of oxidative stress related protein, neuron-restrictive silencer factor, in hippocampal neurons were detected by immunofluorescence histochemistry and western blot assays. After 2 months of drug administration,(1) in the model group, the average escape latency was longer than that of the sham group, and the proportion of straight and tend tactics was lower than that of the sham group, and the hippocampal neurons were irregularly arranged and the cytoplasm was hyperchromatic.(2) The levels of reactive oxygen species and malondialdehyde in the hippocampus of the model group rats were increased, and the activity of superoxide dismutase was decreased.(3) Lycopene(50, 100 and 200 mg/kg) intervention improved the above changes, and the lycopene 100 mg/kg group showed the most significant improvement effect.(4) Neuron-restrictive silencer factor expression in the hippocampus was lower in the sham group and the lycopene 100 mg/kg group than in the model group.(5) The above data indicate that lycopene 100 mg/kg could protect against the learning-memory ability impairment of vascular dementia rats. The protective mechanism was achieved by inhibiting oxidative stress in the hippocampus. The experiment was approved by the Animal Ethics Committee of Fujian Medical University, China(approval No. 2014-025) in June 2014.

Lycopene and male infertility

Excessive amounts of reactive oxygen species （ROS） cause a state of oxidative stress, which result in sperm membrane lipid peroxidation, DNA damage and apoptosis, leading to decreased sperm viability and motility. Elevated levels of ROS are a major cause of idiopathic male factor infertility, which is an increasingly common problem today. Lycopene, the most potent singlet oxygen quencher of all carotenoids, is a possible treatment option for male infertility because of its antioxidant properties. By reacting with and neutralizing free radicals, lycopene could reduce the incidence of oxidative stress and thus, lessen the damage that would otherwise be inflicted on spermatozoa. It is postulated that lycopene may have other beneficial effects via nonoxidative mechanisms in the testis, such as gap junction communication, modulation of gene expression, regulation of the cell cycle and immunoenhancement. Various lycopene supplementation studies conducted on both humans and animals have shown promising results in alleviating male infertility--lipid peroxidation and DNA damage were decreased, while sperm count and viability, and general immunity were increased. Improvement of these parameters indicates a reduction in oxidative stress, and thus the spermatozoa is less vulnerable to oxidative damage, which increases the chances of a normal sperm fertilizing the egg. Human trials have reported improvement in sperm parameters and pregnancy rates with supplementation of 4-8 mg of lycopene daily for 3-12 months. However, further detailed and extensive research is still required to determine the dosage and the usefulness of lycopene as a treatment for male infertility.

Supplemental blue and red light promote lycopene synthesis in tomato fruits

Lycopene, one of the strongest natural antioxidants known and the main carotene in ripe tomato, is very important for human health. Light is well known to be one of the most important environmental stimuli influencing lycopene biosynthesis; specifically, red light induces higher lycopene content in tomato. However, whether blue light promotes lycopene synthesis remains elusive and exactly how light stimulation promotes lycopene synthesis remains unclear. We applied supplemental blue and red lighting on tomato plants at anthesis to monitor the effect of supplemental blue and red lighting on lycopene synthesis. Our results showed that supplemental blue/red lighting induced higher lycopene content in tomato fruits; furthermore, we found that the expression of key genes in the lycopene synthesis pathway was induced by supplemented blue/red light. The expression of light signaling components, such as red-light receptor phytochromes(PHYs), blue-light receptor cryptochromes(CRYs) and light interaction factors, phytochrome-interacting factors(PIFs) and ELONGATED HYPOCOTYL 5(HY5) were up-or down-regulated by blue/red lighting. Thus, blue and red light increased lycopene content in tomatoes by inducing light receptors that modulate HY5 and PIFs activation to mediate phytoene synthase 1(PSY1) gene expression. These results provide a sound theoretical basis for further elucidation of the light regulating mechanism of lycopene synthesis in tomatoes, and for instituting a new generation of technological innovations for the enhancement of lycopene accumulation in crop production.

Hepatoprotective and antioxidant effects of lycopene on non-alcoholic fatty liver disease in rat

AIM To evaluate the hepatoprotective effect of lycopene(Ly) on non-alcoholic fatty liver disease(NAFLD) in rat. METHODS A rat model of NAFLD was first established by feeding a high-fat diet for 14 wk. Sixty-five rats were randomly divided into normal group, model group and Ly treatment groups. Alanine aminotransferase(ALT), aspartate aminotransferase(AST), triglycerides(TG), total cholesterol(TC) in serum and low density lipoproteincholesterol(LDL-C), high density lipoprotein-cholesterol(HDL-C), free fatty acid(FFA), malondialdehyde(MDA), superoxide dismutase(SOD), glutathione(GSH) in liver tissue were evaluated, respectively. While the hepatoprotective effect was also confirmed by histopathological analysis, the expression levels of TNF-α and cytochrome P450(CYP) 2E1 in rat liver were determined by immunohistochemistry analysis.RESULTS A significant decrease was observed in the levels of serum AST(2.07-fold), ALT(2.95-fold), and the blood lipid TG(2.34-fold) and TC(1.66-fold) in the dose of 20 mg/kg Ly-treated rats(P < 0.01), compared to the model group. Pretreatment with 5, 10 and 20 mg/kg of Ly significantly raised the levels of antioxidant enzyme SOD in a dose-dependent manner,to 90.95 ± 9.56, 109.52 ± 11.34 and 121.25 ± 10.68(P < 0.05, P < 0.01), as compared with the model group. Similarly, the levels of GSH were significantly increased(P < 0.05, P < 0.01) after the Ly treatment. Meanwhile, pretreatment with 5, 10 and 20 mg/kg of Ly significantly reduced MDA amount by 30.87, 45.51 and 54.49% in the liver homogenates, respectively(P < 0.01). The Ly treatment group showed significantly decreased levels of lipid products LDL-C(P < 0.05, P < 0.01), improved HDL-C level and significantly decreased content of FFA, compared to the model group(P < 0.05, P < 0.01). Furthermore, the Ly-treated group also exhibited a down-regulated TNF-α and CYP2E1 expression, decreased infiltration of liver fats and reversed histopathological changes, all in a dosedependent manner(P < 0.05, P < 0.01). CONCLUSION This study suggests that Ly has a protective effect on NAFLD, down-regulates expression of TNF-α, and that CYP2E1 may be one of the action mechanisms for Ly.

Tomato lycopene attenuates myocardial infarction induced by isoproterenol:Electrocardiographic,biochemical and anti-apoptotic study

Objective:To assess the protective effects of lycopene on electrocardiographic,hemodynamic, biochemical and apoptotic changes in isoproterenol induced myocardial infarction.Methods: Myocardial infarction was induced in rats by subcutaneous injection of isoproterenol(200 mg/kg) for two consecutive days at an interval of 24 h.Rats were treated with lycopene(10 mg/kg/day, p.o.) for a period of 30 days and isoproterenol(ISO) was injected on the 29th and 30th day.At the end of experiment i.e.on the 31st day electrocardiographic,hemodynamic,biochemical and apoptotic changes were monitored from control and experimental groups.Results:ISO injected ruts showed a significant alteration in electrocardiograph pattern and hemodynamic changes(i.e. systolic,diastolic and mean arterial pressure).It also showed significant increase in C-reactive protein,myeloperoxidase,nitrite levels and Caspase-3 protease activity.In addition,it also exhibited alteration in the levels of electrolytes(Na^+,K^+ and Ca^(2+),vitamin E,uric acid and serum protein.Cel electrophoresis of ISO injected rats showed increase in DNA fragmentation.Triphenyl tetrazolium chloride staining of the heart section shows increase area of infarction in ISO injected rats.Pre-co-treatment with lycopene significantly prevented the ISO induced ullerution in ECC, haemodynamic,biochemical and apoptotic changes.Conclusions:The present result shows that treatment of lycopene in ISO injected rats significantly attenuates induced myocardial infarction.

The Protective Role of Procyanidins and Lycopene Against Mercuric Chloride Renal Damage in Rats

Objective This study aims to investigate the protection of procyanidins and lycopene from the renal damage induced by mercuric chloride.Methods Rats were treated with either procyanidins or lycopene 2h before HgCl 2 subcutaneously injection,once daily treatment for 2 successive days.Results In comparison with HgCl 2 group,markers of renal function such as blood urea nitrogen in serum and urinary protein were decreased to （18.45±11.63） mmol/L and （15.93±9.36） mmol/L,（4.54±0.78） g/（g Cr） and （4.40±1.12） g/（g Cr）.N‐acetyl‐beta‐D‐glucosaminidase,lactate dehydrogenase,alkaline phosphatase in urine were depressed to （125.49±11.68） U/（g Cr）,（103.73±21.79） U/（g Cr）,（101.99±12.28） U/（g Cr）,and （113.19±23.74） U/（g Cr）,（71.14±21.80） U/（g Cr）,（73.64±21.51） U/（g Cr） in procyanidins and lycopene groups.Indicators of oxidative stress,for example,Glutathion was reduced to （45.58±9.89） μmol/（g pro） and （45.33±5.90） μmol/（g pro）,and antioxidant enzymes such as superoxide dismutase,glutathione‐peroxidase were enhanced to （43.07±10.97） U/（mg pro） and （39.94±6.04） U/（mg pro）,（83.85±18.48） U/（mg pro）,and （85.62±12.68） U/（mg pro）.Malondialdehyde was lowered to （0.95±0.12） （μmol/g pro） and （1.03±0.12） μmol/（g pro） in procyanidins and lycopene groups.ROS generation was decreased by 27.63% and 16.40% and apoptosis was also decreased in procyanidins and lycopene groups respectively.Pathological changes were much better as well.Conclusion Procyanidins and Lycopene play some protective role against mercury kidney damage.

Lycopene stabilizes lipoprotein levels during D-galactosamine/lipopolysaccharide induced hepatitis in experimental rats

Objective:To investigate the effect of lycopene on lipoprotein metabolism during D-galactosamine/lipopolysacchoride(D-Gal/LPS)induced hepatitis in experimentul rats.Methods:The efficacy of lycopene was validated during D-Gal/LPS induced hepatitis by analyzing the activity of lipid metabolizing enzymes such as lipoprotein lipase(LPL),lecithincholesterol acyl transferase(LCAT)and hepatic triglyceride lipase(HTCL).Lipo protein analyses were done by the estimation of very low density lipoprotein cholesterol(VLDL),low density lipoprotein cholesterol(LDL)and high density lipoprotein cholesterol(HDL).Remits:The toxic insult of D-galactosamine/lipopolysaccharide(D-Gal/LPS)in experimental group of animals reduces the normal values of lipid metabolizing enzymes due to liver injury.The significant drop in the levels of HDL and concomitant increase in the values of VLDL and LDL were observed.The pretreatment of lycopene restore these altered values to near normal level in experimental group of animals.Conclusions:In the light of results,it can be concluded that administration lycopene stabilizes the lipoprotein levels by regulating the lipid metabolizing enzymes through its antioxidant defense and helps to maintain the normal lipid metabolism during toxic injury in liver.

Tissue differential expression of lycopene β-cyclase gene in papaya

Carotene pigments in flowers and fruits are distinct features related to fitness advantages such as attracting insects forpollination and birds for seed dispersal.In papaya,the flesh color of the fruit is considered a quality trait that correlateswith nutritional value and is linked to shelf-life of the fruit.To elucidate the carotenoid biosynthesis pathway in papaya,we took a candidate gene approach to clone the lycopene β-cyclase gene,LCY-B.A papaya LCY-B ortholog,cpLCY-B,was successfully identified from both cDNA and bacterial artificial chromosome(BAC)libraries and complete genomicsequence was obtained from the positive BAC including the promoter region.This cpLCY-B shared 80% amino acididentity with citrus LCY-B.However,full genomic sequences from both yellow- and red-fleshed papaya were identical.Quantitative real-time PCR(qPCR)revealed similar levels of expression at six different maturing stages of fruits forboth yellow-and red-fleshed genotypes.Further expression analyses of cpLCY-B showed that its expression levels wereseven- and three-fold higher in leaves and,respectively,flowers than in fruits,suggesting that cpLCY-B is down-regulatedduring the fruit ripening process.

Study on the Separation,Extraction of Lycopene and Its Effects on Cell Cycle

The separation, extraction of lycopene and its effects on the proliferation and cells cycle of the chemical-induced cells were investigated in order to research on its extraction method and the mechanism in inhibiting neoplastic transformation. The best extraction condition of lycopene with super-critical carbon dioxide was under the pressure of 25MPa, the temperature of 50℃ and duration of 3. 0h. Lycopene could inhibit cell growth rate and cells proliferation significantly, while increase the cell numbers of G1 -phase and decrease that of S-phase and G2+M-phase. The potency of the effects of lycopene on cells cycle might be one of the important reasons for inhibiting neoplastic transformation.

Two Lycopene β-Cyclases Genes from Sweet Orange (Citrus sinensis L. Osbeck) Encode Enzymes With Different Functional Efficiency During the Conversion of Lycopene-to-Provitamin A

Citrus fruits are rich in carotenoids.In the carotenoid biosynthetic pathway,lycopene β-cyclase（LCYb,EC：1.14.-.-） is a key regulatory enzyme in the catalysis of lycopene to β-carotene,an important dietary precursor of vitamin A for human nutrition.Two closely related lycopene β-cyclase cDNAs,designated CsLCYb1 and CsLCYb2,were isolated from the pulp of orange fruits（Citrus sinensis）.The expression level of CsLCYb genes is lower in the flavedo and juice sacs of a lycopeneaccumulating genotype Cara Cara than that in common genotype Washington,and this might be correlated with lycopene accumulation in Cara Cara fruit.The CsLCYb1 efficiently converted lycopene into the bicyclic β-carotene in an Escherichia coli expression system,but the CsLCYb2 exhibited a lower enzyme activity and converted lycopene into the β-carotene and the monocyclic γ-carotene.In tomato transformation studies,expression of CsLCYb1 under the control of the cauliflower mosaic virus（CaMV） 35S constitutive promoter resulted in a virtually complete conversion of lycopene into β-carotene,and the ripe fruits displayed a bright orange colour.However,the CsLCYb2 transgenic tomato plants did not show an altered fruit colour during development and maturation.In fruits of the CsLCYb1 transgenic plants,most of the lycopene was converted into β-carotene with provitamin A levels reaching about 700 μg g-1DW.Unexpectedly,most transgenic tomatoes showed a reduction in total carotenoid accumulation,and this is consistent with the decrease in expression of endogenous carotenogenic genes in transgenic fruits.Collectively,these results suggested that the cloned CsLCYb1 and CsLCYb2 genes encoded two functional lycopene β-cyclases with different catalytic efficiency,and they may have potential for metabolite engineering toward altering pigmentation and enhancing nutritional value of food crops.

Studies on biological effect of lycopene on Hippocampus of hyperlipemia rats

Objective: This study investigated into the ef-fect of lycopene on expression of APP, bax and bcl-2 in hippocampal CA1 region of rats with hyperlipidemia. Methods: By total cholesterol (TC) and body weight, 48 adult male SD rats were randomized into six groups, a normal control group, fed with basic feed;a high-fat model group, fed with high-fat feed;a positive drug control group, fed with high-fat feed and administrated with fluvastatin sodium at a dose of 10 mg?kg?bw-1?d-1 by gastric perfusion;and lycopene groups at three dose levels, fed with high-fat feed and administrated with lycopene at doses of 11, 22 and 44 mg?kg?bw-1?d-1 respec-tively also by gastric perfusion. Caudal venous blood samples of rats in all groups were taken at week 0, week 1 and week 3 after the experiment started so as to assay TC, TG, LDL-C and HDL-C;at the end of the experiment, rat brains were taken and sections of the hippocampal CA1 re-gion were prepared. Expression of APP, bax and bcl-2 in the CA1 region was determined by im-munohistochemical methods and morphologi-cal examination was carried out. Results: One week after fed with high-fat feed, models of hy-perlipidemia rats were established;at the end of experiment, hippocampal APP and bax expres-sion was enhanced while bcl-2 expression was significantly weakened (p&amp;lt;0.05);to rats with hyperlipidemia, both lycopene and fluvastatin sodium could reduce TC, TG and LDL-C, inhibit expression of hippocampal APP and bax and promote expression of bcl-2 (p&amp;lt;0.05). Conclu- sion: Lycopene down-regulates the expression of bax and up-regulates that of bcl-2 mainly by reducing serum TC and LDL-C and weakening expression of APP in the hippocampal CA1 re-gion of rats with hyperlipidemia, thereby main-taining normal morphology of hippocampal neurons and facilitating the protection of the brain.

Lycopene Ameliorates Diabetic-Induced Changes in Erythrocyte Osmotic Fragility and Lipid Peroxidation in Wistar Rats

Introduction: Diabetes mellitus has remained one of the serious health problems in the world;and oxidative stress has been reported to be a root cause for the progression and development of diabetes mellitus and its associated complications. Aim: This study investigated the possible ameliorative effects of lycopene on diabetic-induced changes in erythrocyte osmotic fragility and lipid peroxidation in Wistar rats. Methodology: The animals were made diabetic by single intraperitoneal injection of streptozotocin at 60 mg/kg b w. Diabetes was confirmed by the presence of high fasting blood glucose level ≥ 200 after 72 hours. Thereafter, diabetic rats were randomly assigned into six groups (1, 2, 3, 4, 5 and 6) comprising five animals each. Group 1 (Diabetic control) and group 2 (Normal control) rats received 0.5 ml of olive oil, groups 3, 4, 5 rats received 10, 20, 40 mg/kg bw of lycopene respectively, while those in group 6 received 2 mg/kg bw of glibenclamide orally once daily for a period of four weeks. At the end of the treatment, all animals were sacrificed;blood samples collected for determination of erythrocyte osmotic fragility (EOF) and lipid peroxidation (LPO). Results: The results obtained showed that there was a significantly (P Conclusion: From the available findings, it can be concluded that administration of lycopene to diabetic rats attenuated diabetic-induced changes in EOF and LPO and these observed effects may be attributed to anti-oxidative property of lycopene.

Lycopene modulates cellular proliferation, glycolysis and hepatic ultrastructure during hepatocellular carcinoma

AIMTo investigate the effect of lycopene extracted from tomatoes (LycT) on ultrastructure, glycolytic enzymes, cell proliferation markers and hypoxia during N-Nitrosodiethylamine (NDEA)-induced hepatocarcinogenesis. METHODSFemale BALB/c mice were randomly divided into four groups: The Control, NDEA (200 mg NDEA/kg b.w. given i.p.), LycT (5 mg/kg b.w. given orally on alternate days) and LycT + NDEA group. The mRNA and protein expression of various cell proliferation markers (PCNA, Cyclin D1, and p21) were assessed by reverse transcription-polymerase chain reaction and enzyme linked immunosorbent assay, respectively. The ultrastructure of hepatic tissue was analyzed using scanning and transmission electron microscopy. The enzymatic activity of glycolytic enzymes was estimated using standardized protocols, while glucose-6-phosphate dehydrogenase activity level was estimated using a kit obtained from Reckon Diagnostic P. Ltd. (India). RESULTSUncontrolled proliferation in the liver of NDEA (P ≤ 0.001) mice was evident from the high expression of cell-proliferation associated genes (PCNA, Cyclin D1, and p21) when compared to control and LycT mice. In addition, enhanced activities of hexokinase, phosphoglucoisomerase, aldolase, glucose-6-phosphate dehydrogenase and hypoxia-inducible factor-1&alpha; were observed in NDEA mice as compared to control (P ≤ 0.001) and LycT (P ≤ 0.001) mice. The alterations in hepatic ultrastructure observed in the NDEA group correlated with the changes in the above parameters. LycT pre-treatment in NDEA-challenged mice ameliorated the investigated pathways disrupted by NDEA treatment. Moreover, hepatic electron micrographs from the LycT + NDEA group showed increased macrophages, apoptotic bodies and well-differentiated hepatocellular carcinoma (HCC) in comparison to undifferentiated HCC as observed in the NDEA treated group. CONCLUSIONThis study demonstrates that dietary supplementation with LycT has a multidimensional role in preventing HCC development.

Effect of ploidy level on expression of lycopene biosynthesis genes and accumulation of phytohormones during watermelon(Citrullus lanatus) fruit development and ripening

The difference between lycopene and phytohormone levels among diploid, triploid and tetraploid plants of two watermelon cultivars during fruit growth and ripening was studied. The expression pattern of five genes（phytoene synthase（PSY1）, phytoene desaturase（PDS）, ζ-carotene desaturase（ZDS）, carotenoid isomerase（CRTISO）, and lycopene β-cyclase（LCYB）） was analyzed in details. In red-fleshed cultivar Mimei, lycopene content increased rapidly from 25 to 35 days after pollination（DAP）, and then decreased at 40 DAP. Triploid and tetraploid fruit had higher levels of lycopene than diploid. Moreover, triploid tended to contain more lycopene than tetraploid during fruit growth and ripening stages. However, little amount of lycopene（0–2 mg kg–1 fresh weight（FW）） in yellow-fleshed cultivar Huangmei was found during all fruit development stages. In Mimei, transcript level of PSY1 was generally higher than the other four genes, and LCYB gene expression was the lowest among all five genes being tested. PSY1, CRTISO and LCYB genes showed higher transcript levels in polyploid than in diploid fruit. By contrast, in Huangmei, transcript level of LCYB was not the lowest, but only lower than that of PSY1. PSY1, CRTISO and LCYB genes showed higher expression levels in diploid than in polyploid fruit. In Mimei, the negative correlation between gibberellane（GA） content and lycopene accumulation was determined in all three different ploidy fruits, while a positive correlation was observed between abscisic acid（ABA） content and lycopene accumulation only in diploid watermelon. These results indicated that different lycopene contents in different ploidy watermelons is regulated by the differential transcription expression of the lycopene metabolic genes and phytohormones.

Antioxidant efficiency of lycopene on oxidative stress-induced damage in bovine spermatozoa

Background： Lycopene（LYC） is a natural carotenoid with powerful reactive oxygen species（ROS） scavenging activities. The aim of this study was to investigate if lycopene has the ability to reverse ROS-mediated alterations to the motility, viability and intracellular antioxidant profile of bovine spermatozoa subjected to ferrous ascorbate（Fe AA）. Spermatozoa were washed out of fresh bovine semen, suspended in 2.9 % sodium citrate and subjected to LYC treatment（0.25, 0.5, 1 or 2 mmol/L） in the presence or absence of Fe AA（150 μmol/L Fe SO4 and 750 μmol/L ascorbic acid） during a 6 h in vitro culture. Spermatozoa motion characteristics were assessed using the Sperm Vision？computer-aided sperm analysis（CASA） system. Cell viability was examined with the metabolic activity（MTT） assay,ROS generation was quantified via luminometry and the nitroblue-tetrazolium（NBT） test was applied to quantify the intracellular superoxide formation. Cell lysates were prepared at the end of the in vitro culture to investigate the intracellular activity of superoxide dismutase（SOD）, catalase（CAT）, glutathione peroxidase（GPx） as well as the concentrations of glutathione（GSH） and malondialdehyde（MDA）.Results： FeA A treatment led to a reduced spermatozoa motility（P 〈 0.001）, viability（P 〈 0.001） and a decline of the antioxidant capacity of spermatozoa（P 〈 0.001） but increased the ROS generation（P 〈 0.001）, superoxide production（P 〈 0.001） and lipid peroxidation（P 〈 0.001）. LYC administration resulted in a preservation of the spermatozoa motion parameters（P 〈 0.001）, mitochondrial activity（P 〈 0.001） and antioxidant characteristics（P 〈 0.001 with respect to SOD;P 〈 0.01 in relation to CAT; P 〈 0.05 as for GPx and GSH） with a concentration range of 1 and 2 mmol/L LYC revealed to be the most effective.Conclusions： Our results suggest that LYC exhibits significant ROS-scavenging and antioxidant properties which may prevent spermatozoa alterations caused by oxidative stress, and preserve the functionality of male reproductive cells.