AMELIORATING THE METABOLIC DISORDER IN STREPTOZOTOCIN-INDUCED DIABETIC RATS BY HAEMATOCOCCUS PLUVIALIS EXTRACT

Objective: The purpose of the current study is to evaluate the antidiabetic influence of Haematococcus pluvialis (H. pluvialis) ethanolic extract on streptozotocin (STZ)-induced rats.Methods: This influence was assessed by measuring; asymmetric dimethylarginine (ADMA), von willebrand factor (vWf), 15-lipoxygenase (15-LOX), 5-lipoxygenase (5-LOX), epidermal growth factor (EGF) and lipoxin-A4 (LXA4). The histopathological examinations of cardiac and renal tissues were also performed.Results: The presented  results showed an elevation in the levels of ADMA, vWf, 15-LOXand 5-LOX with percentage increase  430.30, 77.54, 61.05 and 81.09 %, respectively in diabetic rats. However, the EGF and LXA4 levels were decreased significantly (44.17 and51.94%, respectively) compared to normal control rats. On the other hand, treatment of diabetic rats with H. pluvialis improved ADMA, vWf, 15-LOX, 5LOX, EGF and LXA4 levels with amelioration percentages; 269.69, 54.77, 55.78, 72.68, 53.39 and 56.58%, respectively. Additionally, the histopathological examination of diabetic cardiac tissue revealed congestion of myocardial blood vessel, vacuolation of the sarcoplasm of cardiac myocytes and focal necrosis of myocytes associated with inflammatory cells infiltration. Beside, vacuolation of epithelial lining of renal tubules and endothelial lining glomerular tuft as well as cystic dilatation of renal tubules were detected. However, cardiac tissue  and renal tubules of diabetic rats treated with H. pluvialis ethanolic extract (150 mg/kg b.wt.) showed normal cardiac myocytes, vacuolation of epithelial lining renal tubules and endothelial lining glomerular tuft, congestion of glomerular tuft and intertubular renal blood vessels.Conclusion: These results confirmed the improving role of H. pluvialis in controlling type 2diabetes.Â

Antioxidant and AntiproliferativeEffects on Human Liver HePG2Epithelial Cells from Artichoke (Cynara scolymus L.) By-Products

Cynara scolymus L.is a medicinal plant frequently used in traditional medicine for liver diseases. This study aimed to evaluate the antioxidant and Anti-tumer effect in human Liver HePG2 epithelial cells of artichoke. The antioxidant activity of the free phenolic of bracts methanolic extract, as measured with the DPPH.freeradical scavenging activity and iron chelating showed significant antioxidant activity > 0.05  as well as yielded the largest quantity of  phenolics and flavonoids  compounds. A significant increase was observed in hepatocellular carcinoma cell line (HePG2) 78.3 % treated with free phenolic extract of bracts higher than that found in by the heart of artichoke 36.7%. This work showed that extracts with high antioxidant activity can be obtained from artichoke and artichoke by-products. Therefore, this study indicate that the free phenolic extract from bracts of Cynara scolymusL. might be of interest within the developing market of nutritional ingredients and is capable of yielding nutritional supplements with antioxidant and anticancer activities. Key words: Artichoke by-products, antioxidant, antitumor, HepG

A Comparative Study on the Active Constituents, Antioxidant Capacity and Anti-Cancer Activity of Cruciferous Vegetable Residues

أجريت هذه الدراسة لتقييم كفاءة وفعالية مخلفات أربعة أنواع من خضروات العائلة الصليبية (البراسيكا) تم ستخلاصهم بمذيبين (الميانولي  - المائي) وهم كاتالي البروكولي ، كرنب بروكسل ، القرنبيط ، الفجل الأحمر. وتم تقييم المستخلصات من حيث محتواها الكلي من المركبات الفعالة مثل الفينولات و الفلاڤونيدات والتانينات وڤيتامين سي واتعرف عليها بالـ HPLC ودراسة نشاطاتها المضادة للأكسدة بأكثر من طريقة مثل الـ DPPH· والـ Fe2+-chelating والـ Reducing power وكذلك دراسة كفاءتها في كبح جماح ومنع إنتشار وتكاثر ثلاثة أنواع من الخلايا السرطانية للإنسان علي النطاق المعملي مثل خلايا سرطان الكبد HePG2 و خلايا سرطان القولون HTC116 وخلايا سرطان الثدي MCF7. أظهرت النتائج أن المستخلص الميثانولي لمخلفات البروكولي وكرنب بروكسيل أعطي أعلي محتوي من المركبات الفعالة و أعلي نشاط مضاد للأكسدة في كبح جماح الشق الحر الـ DPPH· و وأعلي قدرات إختزلية ضد الحديديك الـ Reducing power. بينما أظهر المستخلص المائي لكل من كرنب بروكسيل والقرنبيط اعلى نشاط في عملية خلب الحديدوز الـ Fe2+-chelating. أكد تحليل HPLC للتعرف علي المركبات الفينولية الموجودة في مستخلصات مخلفات خضروات البراسيكا التي تم التعرف عليها أن المستخلص الميثانولي يمتلك كميات مرتفعة من الكاتشين والروتين والكيوماريك والبنزويك و الليتيولين. كما أظهر المستخلص الميثانولي لمخلفات كرنب بروكسيل تأثيراً مثبطاً و مانع للإنتشار وقاتل مع كافة أنواع الخلايا السرطانية وكان ذلك متبوعاً بالبروكولي والفجل الاحمر على التوالي. أكدت هذه النتائج أن مخلفات كرنب بروكسيل تحتوي على مركبات كيميائية فعالة يمكنها أن تمنع تكاثر الخلايا السرطانية. لذلك ، اقترحت هذه النتائج أن مخلفات خضروات البراسيكا يمكن إستخدامها كمضادات أكسدة قوية ومضادة لإنتشار الخلايا السرطانية وذلك في تطبيقات الصناعات الغذائية و الدوائية.This study is pointed out to estimate the effectiveness of two solvents in the extraction and evaluating the active ingredients and their antioxidant activity as well as anti-cancer efficiency. Therefore, residues from four different Brassica vegetables viz. broccoli, Brussels sprout, cauliflower, and red cherry radish were extracted using two procedures methods: methanolic and water crude extracts. Methanol extracts showed the highest content of total phenolic (TP), total flavonoids (TF), and total tannins (TT) for broccoli and Brussels sprouts residues. Methanolic extract of broccoli and Brussels sprouts residues showed the highest DPPH· scavenging activity (IC50 = 15.39 and 18.64 µg/ml). The methanol and water extracts of Brussels sprout residues showed the highest chelating activity (IC50 = 11.77 and 5.94 µg/ml) and exhibited the highest reducing power (EC50 =14.38 and 20.18 µg/ml) with broccoli respectively. The HPLC analysis of phenolic compounds confirmed that the methanol extract of all the residues examined possessed high amounts of catchine, rutin, cumaric, benzoic, and luteolin. The methanol extract at 100 µg/ml of Brussels sprouts residues displayed a rise cytotoxic effect on HePG2 (80.40%), MCF7 (75.49%) and HCT116 (22.74%) followed by broccoli and red cherry radish, respectively. This result confirmed that Brussels sprouts residue contain effective chemical compounds that can inhibit the proliferation of cancer cells. Therefore, these results proposed that those Brassica vegetable residues might be beneficial as a potent antioxidant and anticancer agents and strongly recommended as fixing in constituent's food applications and pharmaceutical industries

BERRY EXTRACTS IMPROVED INFLAMMATORY CYTOKINES, ANTIOXIDANT ENZYME AND SUPPRESSED THE GENE EXPRESSION ALTERATIONS IN DIABETIC RATS

Objective: The present work was conducted to evaluate the ameliorative effect of red berry (Morus rubra) and white berry (Morus alba) extracts in diabetic rats.Methods: Inflammatory biomarkers; tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), transforming growth factor-β (TGF-β), paraoxanase1 (PON1), gene expression of apoptosis and inflammatory related genes were carried out.Results: Treatment of male rats with streptozotocin (STZ) to induce diabetes increased the levels of TNF-α, IL-1β, TGF-β, the apoptosis rates and the alterations of the pro-apoptosis (eotaxin, caspase-1, and caspase-2) and inflammation; nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB1) and allograft inflammatory factor 1 (AIF-1) related genes. While marked reduction in PON1 level was detected in STZ-induced diabetic rats. However, treatment of STZ-exposed rats with red and white berry extracts exhibited noticeable ameliorations in TNF-α, IL-1β, TGF-β levels and PON1 activity as well as low apoptosis rates and decreased the alteration of the pro-apoptosis and inflammatory related genes.Conclusion: Red and white berry extracts exhibited attenuation in antioxidant status, suppressed inflammatory cytokines, apoptosis and the alterations in the apoptosis and inflammatory related genes induced by STZ in male rats

HEALING POTENCY OF HAEMATOCOCCUS PLUVIALIS EXTRACT FOR TREATING TYPE 2 DIABETES IN RATS

Objective: The present study aims to evaluate the antidiabetic effect of ethanolic extract of Haematococcus pluvialis (H. pluvialis) in streptozotocin (STZ)-induced diabetic rats.Methods: The antidiabetic activity of H. pluvialis was investigated by the determination of glucose and insulin levels, aspartate (AST), alanine transaminases (ALT), lipid profile including total cholesterol (TC), triglycerides (TG), low-density lipoprotein-cholesterol (LDL-C) and high-density-lipoprotein-cholesterol (HDL-C). Histopathological examination of pancreas and liver were also carried out.Results: The results revealed that the levels of glucose, TC, TG, LDL-C as well as AST and ALT enzyme activities were increased significantly in diabetic rats. While, insulin and HDL-C levels decreased significantly in STZ-induced diabetic rats. The remediation of diabetic rats with H. pluvialis attenuated the elevated levels of glucose, TC, TG, LDL-C as well as AST and ALT activities in diabetic rats. Besides, it improved insulin, HDL-C levels, pancreas and hepatic architectures.Conclusion: H. pluvialis extract has a promising antidiabetic potency through attenuation of several metabolic disorders associated diabetes

STEROLS BIOACTIVITY OF RUTA GRAVEOLENS L. AND MURRAYA PANICULATA L.

Objective: Ruta graveolens L. (R. graveolens) and Murraya paniculata L. (M. paniculata) are medicinal plants belonging to Rutaceae family have many uses in traditional medicine. The aim of the present study was to investigate sterols bioactivity of the two Rutaceae plant leaves.Methods: Sterols of the two Rutaceae plant leaves were identified using GC/MS. The antioxidant activities of the sterols of these herbs were evaluated by three different methods; free radical scavenging using 2,2′-Azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), 1,1-Diphenyl-2-picryl-hydrazyl (DPPH) and total antioxidant activity. The anticancer activity of the sterols was determined by MTT assay against colorectal cancer HCT116, breast cancer MCF7, liver cancer HepG2 and lung cancer A549 cell lines. Anti-inflammatory activity was evaluated using albumin denaturation assay and antiviral activities against H5N1 virus were carried out using plaque reduction assay.Results: GC/MS assay showed β-Sitosterol (36%) as the most abundant sterols of R. graveolens followed by stigmasterol (18%), while stigmasterol (25.2%) was the most abundant one of M. paniculata steroids. The anti-inflammatory potential of R. graveolens steroids was significantly higher than that of diclofenac sodium (standard drug). M. paniculata sterols have higher antiviral activity (IC50= 0.15 of µg/ml) than R. graveolens sterols (IC50= 7.8 of µg/ml). The sterols of R. graveolens showed anticancer activity against MCF7 and A549 cells with inhibition 84.3 and 81%, at 100 µg/ml respectively. While M. paniculata sterols showed 77.3% inhibition against A549 cells.Conclusion: The current study suggests that the sterols of M. paniculata have more anti-viral activity than R. graveolens sterols which showed more anticancer and anti-inflammatory activities

NEURORESTORATIVE MULBERRIES POTENTIAL OF ALZHEIMER'S DISEASE IN ANIMAL MODEL

Objective: Alzheimer's disease (AD) is one of the most common neurodegenerative disorders. The present study aimed to identify the potential neurorestorative of Morus alba and Morus rubra fruits extract on aluminum chloride (AlCl3)-induced AD rats.Methods: Each Morus species extract M. alba and M. rubra at a dose 300 mg/kg body weight (b.wt.) was orally administered, daily for 6 weeks post AlCl3 induction (100 mg/kg b.wt./day/6 weeks, orally). The polyphenols content was determined according to the colorimetric method of Folin–Ciocalteu using gallic acid as a standard. In addition, brain neurotransmitters, total protein content (TPC), and antioxidant biomarkers were determined. Histopathological assessment was performed on the brains of rats.Results: Administration of AlCl3 neurotoxicity induced a significant increment of brain acetylcholinesterase, and nitric oxide, while significant decrease in four monoamines (norepinephrine, epinephrine, 5-hydroxytryptamine serotonin, and dopamin), along with brain tissue ATPase, and TPC were determined. AlCl3-induced changes in peroxidative products, such as protein carbonyl, catalase, and superoxide dismutase activity. AlCl3 induction resulted in significant decrease in antioxidant enzymes, serum level of total antioxidant capacity, and caused histological alteration for brain tissue. Treatment of AD-rats with M. alba and M. rubra (300 mg/kg b.wt./day) causes modulation of most biochemical markers brain function and confirmed by brain histopathological study.Conclusion: Collectively, the present results indicated that M. rubra and M. alba exerts neuroprotection against AlCl3-induced brain pathology. To the best of our knowledge, none of the literature studies have demonstrated the effect of these Morus species against Al-neurotoxicity at biochemical and electron microscopic levels

PHYTOCHEMICAL ANALYSIS, ASSESSMENT OF ANTIPROLIFERATIVE AND FREE RADICAL SCAVENGING ACTIVITY OF MORUS ALBA AND MORUS RUBRA FRUITS

Objective: Mulberry is a nontoxic commonly eaten plant, belongs to the Morus and used in folk medicine in the remedy of dysentery, antiphlogistic, diuretic, expectorant, and antidiabetic. The purpose of this study is to evaluate the antiproliferative and radical scavenging activity of the total alcoholic and successive fractions thereof of Morus alba and Morus rubra fruits. In addition, the chemical composition of the bioactive fractions of each species was investigated.Methods: The antiproliferative potential of 8 extracts on 4 human cancer cell lines, hepatocellular carcinoma (HepG2), Caucasian breast adenocarcinoma (MCF7), prostate (PC3), and colon carcinoma (HCT116) in addition to one normal cell line namely human normal immortalized skin fibroblast cells (BJ1) were carried out. Cell viability was determined using MTT assay. The potency was compared with the reference drug doxorubicin. These extracts were also assayed for 1,1-diphenyl-2-hydrazyl free radical scavenging activities. After saponification of the n-hexane fraction, unsaponifiable matter and fatty acid methyl esters were analyzed by gas liquid chromatography (GLC). The chemical composition of the bioactive fractions was investigated using gas chromatography/mass spectrometry (GC/MS) analysis.Results: All the extracts showed significant free radical scavenging activity dose-dependently. The n-hexane and dichloromethane (DCM) fractions of M. rubra exhibited potent cytotoxic activity on almost cancer cell lines. In the same pattern, ethyl acetate (EtOAc) of M. rubra has moderate cytotoxic activity against all cell lines except HepG2. DCM fraction of M. alba possessed both radical scavenging and high potential antiproliferated activities against HCT116 and MCF7 with inhibitory concentration of 43.9 and 32.3 μg/ml, respectively, while it showed no cytotoxic effect on BJ1. GLC analysis showed the major hydrocarbons in M. alba and M. rubra were heptacosane and docosane, respectively. Sterols were similar in both species but with different ratios and cholesterol was the major one. Palmitic and margaric were the major saturated fatty acid while arachidonic was the major unsaturated fatty acid in both species. GC/MS analysis showed the main compound in DCM fraction of each Morus species was palmitic acid. Furthermore, 1,11-bis-(methoxycarbonyl-ethenyl)-10,2-dihydroxy-cycloeicosane and linolelaidic acid, methyl ester were the main compounds in the EtOAc fraction of each Morus species. Whereas, the main compounds in alcoholic extract of M. alba and M. rubra were methyl-14-methyl-pentadecanoate and 1,2-O-isopropylyidene-4-nonene-1,2,3-triol, respectively.Conclusions: The results observed remarkable biological activity of the successive fractions of M. rubra more than those of M. alba and confirmed its importance as a natural bioactive source. Morus species are good candidates to be promising as possible sources for future antitumor and antioxidants in food and pharmaceutical formulations. The strong activity partly explains the potential effects of Morus species for the treatment of cancer and degenerative diseases caused by free radicals

ANTINEURODEGENERATIVE ACTIVITY OF MICROALGAE DUNALIELLA SALINA IN RATS WITH ALZHEIMER'S DISEASE

Objective: The present study is aimed to investigate the promising action of Dunaliella salina extract as a natural protector against Alzheimer's disease (AD) and reported to possess a variety of activities, including antioxidant effects due to its ability to create large amount of carotenoids.Methods: D. salina is a type of halophile green microalgae was used in the present study. 50 male rats were used in this study, where aluminum chloride was orally administered to induce AD in a dose of 100 mg/kg, daily for 6 weeks. Al-intoxicated rats treated orally daily with D. salina ethanolic extract for 6 weeks in a dose of 150 mg/kg b.wt., whereas standard anti-Alzheimer drug donepezil tartrate was administered at the dose of 10 mg/kg b.wt./day for 6 consecutive weeks. The anti-Alzheimer properties of D. salina extract were achieved through measuring the calmodulin (CaM) level, paraoxonase 1 (PON1) activity, the antiapoptotic marker (Bcl2), brain-derived neurotrophic factor (BDNF), the generation of the DNA adducts (8-hydroxy-2-deoxyguanosine [8-OHdG]/2-deoxy guanosine [2-dG]), and alteration in the expression of amyloid precursor protein, β-site APP-cleaving enzyme 1 (BACE1), and β-site APP-cleaving enzyme 2 (BACE2) in AD rats.Results: The current results demonstrated that supplementation of AD rats with D. salina extract-enhanced CaM level, and increased PON1 activity, upregulated Bcl2 and BDNF, decreased the levels of DNA adducts (8-OHdG/2-dG), and suppressed the alterations of the expression levels of APP, BACE1, and BACE2-m RNAs as compared with those in AD rats.Conclusion: It could be concluded that the biological activity of D. salina extract might be regulated by 9-cis b-carotene protecting the brain cells from the oxidative stress in AD rats.Keywords: Dunaliella salina, Calmodulin, Paraoxonase 1, Bcl2, Brain-derived neurotrophic factor, Alzheimer's disease, DNA adduct, Amyloid precursor protein

HAEMATOCOCCUS PLUVIALIS EXTRACT PROMOTING THE RECOVERY OF MEMORY IMPAIRMENT IN ALZHEIMER'S RATS: ANTI-INFLAMMATORY AND ANTIAPOPTOTIC EFFECTS

ABSTRACTObjective: The present study was conducted to investigate the role of Haematococcus pluvialis extract against oxidative damage, the inflammatory,and apoptotic impacts characterizing the neurodegenerative disorders.Methods: Oxidative stress, B-cell lymphoma 2, brain-derived neurotrophic factor, the inflammation, apoptotic and antiapoptotic impacts in Alzheimer'sdisease (AD) rats were determined through assessment of glutathione reduced (GSH), GSH peroxidase (GPx), lipid peroxide (malondialdehyde), thecytokines level such as tumor necrosis factor-alpha (TNF-α), interleukins (IL-6 and IL-1β), and macrophage inflammation protein (MIP1α) in AD rats.Moreover, the expression of phosphoinositide 3-kinase (PI3K) and serine-threonine protein kinase (Akt) genes regulating the apoptosis in AD ratswas measured.Results: The results revealed that levels of TNF-α, IL-6, IL-1β, and MIP1α were significantly increased in AD rats. Moreover, the expression of PI3Kand Akt genes was downregulated which it was coincided with the increase of apoptosis in AD rats. On the other hand, treatment of AD rats withH. pluvialis extract decreased the oxidative stress of AD in the form of prevention the inflammatory and apoptotic impacts.Conclusion: H. pluvialis could be used for ameliorating AD due to its role in decreases the oxidative stress of AD in the form of prevention theinflammatory and apoptotic impacts. H. pluvialis is a very attractive candidate for uses against neurodegenerative disorders that are caused byincreases oxidative stress inducing neuroinflammation and apoptosis.Keywords: Haematococcus pluvialis, Oxidative stress, Inflammation biomarkers, Apoptotic and antiapoptotic impacts