Antiproliferative and apoptotic effects of proteins from black seeds (Nigella sativa) on human breast MCF-7 cancer cell line.

PMID: 

BMC Complement Med Ther. 2020 Jan 13 ;20(1):5. Epub 2020 Jan 13. PMID: 32020890

Abstract Title: 

Antiproliferative and apoptotic effects of proteins from black seeds (Nigella sativa) on human breast MCF-7 cancer cell line.

Abstract: 

BACKGROUND: Nigella sativa (NS), a member of family Ranunculaceae is commonly known as black seed or kalonji. It has been well studied for its therapeutic role in various diseases, particularly cancer. Literature is full of bioactive compounds from NS seed. However, fewer studies have been reported on the pharmacological activity of proteins. The current study was designed to evaluate the anticancer property of NS seed proteins on the MCF-7 cell line.METHODS: NS seed extract was prepared in phosphate-buffered saline (PBS), and proteins were precipitated using 80% ammonium sulfate. The crude seed proteins were partially purified using gel filtration chromatography, and peaks were resolved by SDS-PAGE. MTT assay was used to screen the crude proteins and peaks for their cytotoxic effects on MCF-7 cell line. Active Peaks (P1 and P4) were further studied for their role in modulating the expression of genes associated with apoptosis by real-time reverse transcription PCR. For protein identification, proteins were digested, separated, and analyzed with LC-MS/MS. Data analysis was performed using online Mascot, ExPASy ProtParam, and UniProt Knowledgebase (UniProtKB) gene ontology (GO) bioinformatics tools.RESULTS: Gel filtration chromatography separated seed proteins into seven peaks, and SDS-PAGE profile revealed the presence of multiple protein bands. Among all test samples, P1 and P4 depicted potent dose-dependent inhibitory effect on MCF-7 cells exhibiting ICvalues of 14.25 ± 0.84 and 8.05 ± 0.22 μg/ml, respectively. Gene expression analysis demonstrated apoptosis as a possible cell killing mechanism. A total of 11 and 24 proteins were identified in P1 and P4, respectively. The majority of the proteins identified are located in the cytosol, associate withbiological metabolic processes, and their molecular functions are binding and catalysis. Hydropathicity values were mostly in the hydrophilic range.CONCLUSION: Our findings suggest NS seed proteins as a potential therapeutic agent for cancer. To our knowledge, it is the first study to report the anticancer property of NS seed proteins.

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The protective effect of Nigella sativa extract on lung inflammation and oxidative stress induced by lipopolysaccharide.

PMID: 

J Ethnopharmacol. 2020 Feb 6 ;253:112653. Epub 2020 Feb 6. PMID: 32035219

Abstract Title: 

The protective effect of Nigella sativa extract on lung inflammation and oxidative stress induced by lipopolysaccharide in rats.

Abstract: 

ETHNOPHARMACOLOGICAL RELEVANCE: Oxidative stress during inflammation can increase inflammation and damage tissue. Nigella sativa L. (NS) showed many pharmacological properties including antioxidant and anti-inflammatory activities.AIM OF THE STUDY: In this study, the preventive effect of NS on lung inflammation and oxidative stress induced by lipopolysaccharide (LPS) in the rats was investigated.MATERIALS AND METHODS: Male rats were assigned to: Control, LPS (1 mg/kg, i.p.), LPS + NS (100, 200, 400 mg/kg, i.p.), (10 per group). Saline (1 ml/kg) was intra-peritoneal (i.p.) injected instead of LPS in the rats of the control group. LPS dissolved in saline and injected i.p. daily for 14 days. Treatment with NS extracts started two days before LPS administration and treatment continued during LPS administration. White blood cells (WBC), total and differential as well as oxidative stress index in bronchoalveolar fluid (BALF) and serum, TGF-β1, IFN-γ, PGE, and IL-4 levels in the BALF and lung histopathology were examined.RESULTS: LPS administration increased total WBC, eosinophils, neutrophils, basophils, and monocytes counts as well as oxidative stress markers in the BALF and serum as well as TGF-β1, IFN-γ, PGE, IL-4 levels in the BALF and pathological changes of the lung tissue. All of these effects were reduced by NS extract treatment dose-dependently.CONCLUSION: These results suggested the protective effects of NS extract on lung inflammation and oxidative stress as well as its effect on lung pathology induced by LPS dose-dependently.

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Dietary supplementation of Morus nigra L. leaves decrease fat mass partially through elevating leptin-stimulated lipolysis.

PMID: 

J Ethnopharmacol. 2020 Mar 1 ;249:112416. Epub 2019 Nov 19. PMID: 31756448

Abstract Title: 

Dietary supplementation of Morus nigra L. leaves decrease fat mass partially through elevating leptin-stimulated lipolysis in pig model.

Abstract: 

ETHNOPHARMACOLOGICAL RELEVANCE: Mulberry leaves are the dry leaves of Morus nigra L. trees, which are widely cultivated in central and southern China. Mulberry has a long history of medicinal use, such as anti-stress, lowering blood glucose and anti-obesity.AIM OF THE STUDY: Explore the effects of mulberry leaves on fat deposition as well as the underlying mechanisms.MATERIALS AND METHODS: Total of 48 fattening pigs weighing about 70 kg were randomly allotted to normal diet or die supplemented with 5% (w/w) mulberry leave powder. Changes of fat mass, indicated by backfat thickness was measured with Piggyback tester, blood triglyceride and cholesterol were tested using commercial biochemical kits, serum hormones were estimatedby ELISA, and leptin-related signaling activity were assessed using western-blot.RESULTS: Supplementation with Mulberry leaf feed (MF) significantly reduced serum triglyceride and free cholesterol concentrations and increased the ratio of high-density lipoprotein cholesterol (HDL-c) to low-density lipoprotein cholesterol (LDL-c), while serum glucose and free fatty acids remained unchanged. Dietary MF resulted in a significant reduction in the size of adipocytes and backfat thickness (P 

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The effect of vitamin D on restless legs syndrome: prospective self-controlled case study.

PMID: 

Sleep Breath. 2020 Feb 20. Epub 2020 Feb 20. PMID: 32078093

Abstract Title: 

The effect of vitamin D on restless legs syndrome: prospective self-controlled case study.

Abstract: 

PURPOSE: To evaluate the effect of vitamin D on severity of restless legs syndrome in patients with idiopathic restless legs syndrome (RLS).METHODS: Patients with idiopathic RLS completed questionnaires including the International Restless Legs Severity Scale (IRLSS) and were evaluated for vitamin D deficiency. Patients with deficiency of vitamin D were treated with 50,000 units per week for 2 months. At the end of the 2 months, vitamin D levels were re-measured and disease severity was re-evaluated in patients who reached adequate vitamin D level. Subgroups of IRLSS questionnaire were also analyzed.RESULTS: Of 35 patients enrolled, 21 (60%) had vitamin D deficiency and received vitamin D therapy. In 2 patients, vitamin D levels did not rise to sufficient levels with supplementation and these 2 patients were excluded from analysis. The remaining 19 patients showed vitamin D levels increased from 13.2± 4.0 to 42.8 ± 9.6 ng/mL while IRLSS improved from 24.9 ± 5.1 to 21.1 ± 2.9 points (p

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Morus nigra leaves extract revokes the depressive-like behavior, oxidative stress, and hippocampal damage induced by corticosterone.

PMID: 

Behav Pharmacol. 2020 Feb 7. Epub 2020 Feb 7. PMID: 32040015

Abstract Title: 

Morus nigra leaves extract revokes the depressive-like behavior, oxidative stress, and hippocampal damage induced by corticosterone: a pivotal role of the phenolic syringic acid.

Abstract: 

The pathophysiology of depression includes glucocorticoids excess, glutamatergic excitotoxicity, and oxidative stress impairment. Previous study demonstrated Morus nigra L. leaves extract and syringic acid (4-hydroxy-3,5-dimethoxybenzoic acid), its major phenolic compound, administered orally for 7 days, decreased the immobility time in the tail suspension test, without locomotor alteration. Therefore, the purpose of this study was to investigate the antidepressant-like effects, antioxidant effects, and neuroprotective effects of M. nigra leaves extract and syringic acid in an animal model of depression induced by corticosterone. Herein, corticosterone administered in male Swiss mice, 60-90 days of age, at 20 mg/kg, once a day, for 21 days, was effective to induce depressive-like phenotype. This alteration was accompanied by the increase of oxidative stress markers (lipid peroxidation, nitrite, and protein carbonyl) and the decrease in nonprotein thiols level, besides impairmentin the hippocampus. Conversely, the treatment with M. nigra leaves extract (10 mg/kg), syringic acid (1 mg/kg), or fluoxetine (10 mg/kg), administered once a day for the last 7 days of the corticosterone treatment, was able to abolish the behavioral alterations elicited by corticosterone, reinforcing evidence of the M. nigra leaves extract and syringic acid having antidepressant-like effect. Both treatments also exerted antioxidant property in the mice's brain, reducing the amount of oxidative stress and abolishing the corticosterone-induced damage in the hippocampal slices. In addition, the treatments protected the hippocampus against the damage induced by the association between corticosterone administration and glutamate excess. In conclusion, M. nigra leaves extract and syringic acid revoke depressive-like behavior induced by corticosterone via inhibition of oxidative stressand hippocampal damage.

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Thymoquinone can be developed as a potential therapeutic modularity in bleomycin-induced lung fibrosis for human use.

PMID: 

Exp Lung Res. 2020 Feb 13:1-11. Epub 2020 Feb 13. PMID: 32053036

Abstract Title: 

Thymoquinone treatment modulates the Nrf2/HO-1 signaling pathway and abrogates the inflammatory response in an animal model of lung fibrosis.

Abstract: 

The present study investigates the therapeutic potential of thymoquinone (TQ) in bleomycin-induced lung fibrosis (BMILF) and elucidates the target-signaling pathway for its effect. Lung fibrosis was induced in rats by a single intra-tracheal instillation of bleomycin (BM) (6.5 U/kg) followed by thymoquinone treatment (10 and 20 mg/kg p.o.) for 28 days. Control rats received saline instead of TQ. Changes in body weight, inflammatory cells count, cytokines levels, and biochemical parameters of the broncho-alveolar lavage fluid (BALF) were recorded. In addition, a histopathology examination and western blotting were performed on lung tissues. BM administration resulted in a significant weight loss, which was ameliorated by TQ treatment. BMILF was associated with a reduction in the antioxidant mechanisms and increased lipid peroxidation. Furthermore, elevated levels of inflammatory cytokines, MMP-7 expression, apoptotic markers (caspase 3, Bax, and Bcl-2), and fibrotic changes including TGF-β and hydroxyproline levels in lung tissues were evident. These abnormalities were diminished with TQ treatment. Likewise, altered total and differential cell count inBALF was significantly improved in rats treated with TQ. TQ also produced a dose-dependent reduction in the expressions of Nrf2, Ho-1 and TGF-β. These results propose that the Nrf2/Ho-1 signaling pathway is a principal target for TQ protective effect against BMILF in rats. Furthermore, TQ decreasesinflammatory oxidative stress possibly through the modulation of nuclear factor Kappa-B (NF-κB) and thereby minimization of collagen deposition in the lung. Therefore, TQ can be developed as a potential therapeutic modularity in BMILF for human use.

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Astragaloside IV suppresses inflammatory response via suppression of NF-κB, and MAPK signalling in human bronchial epithelial cells.

PMID: 

Arch Physiol Biochem. 2020 Feb 14:1-10. Epub 2020 Feb 14. PMID: 32057253

Abstract Title: 

Astragaloside IV suppresses inflammatory response via suppression of NF-κB, and MAPK signalling in human bronchial epithelial cells.

Abstract: 

Astragaloside IV isolated from(Fisch.), which was reported to have anti-tumor, anti-asthma, and suppressed cigarette smoke-induced lung inflammation in mice.This study investigated whether astragaloside IV reduced the expression of inflammatory mediators and oxidative stress in BEAS-2B cells.BEAS-2B cells treated with astragaloside IV, and then stimulated with TNF-α or TNF-α/IL-4. The levels of cytokine and chemokine were analysed with ELISA and real-time PCR.Astragaloside IV significantly inhibited the levels of CCL5, MCP-1, IL-6 and IL-8. Astragaloside IV also reduced ICAM-1 expression for blocked THP-1 monocyte adhesion to BEAS-2B cells. Furthermore, astragaloside IV attenuated the phosphorylation of MAPK, and reduced the translocation of p65 into the nucleus. Astragaloside IV could increase the expression of HO-1 and Nrf2 for promoting the oxidant protective effect.Aastragaloside IV has an anti-inflammatory and oxidative effect via regulated NF-κB, MAPK and HO-1/Nrf2 signalling pathways in human bronchial epithelial cells.

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Total glucosides of paeony decreases apoptosis of hepatocytes and inhibits maturation of dendritic cells in autoimmune hepatitis.

PMID: 

Biomed Pharmacother. 2020 Apr ;124:109911. Epub 2020 Jan 28. PMID: 32004939

Abstract Title: 

Total glucosides of paeony decreases apoptosis of hepatocytes and inhibits maturation of dendritic cells in autoimmune hepatitis.

Abstract: 

Total glucosides of paeony (TGP), an active mixture extracted from paeony root, has anti-inflammatory and immunoregulatory effects and is widely used for the treatment of autoimmune diseases such as rheumatoid arthritis. However, the role of TGP in autoimmune hepatitis (AIH) is still unknown. In this study, we aimed to investigate the effect of TGP in autoimmune liver disease (AILD) patients and in concanavalin A (Con A)-induced experimental autoimmune hepatitis (EAH). Changes in biochemical parameters of AILD patients showed that treatment with TGP exerts significant protective effects on liver function, as reflected by decreased levels of serum alanine transaminase, aspartate transaminase,γ-glutamyl transpeptidase and total bilirubin. In EAH mice, we found that pretreatment with TGP reduced the levels of serum liver enzyme levels, histopathological damage and hepatocyte apoptosis. Importantly, flow cytometry analysis showed that pretreatment with TGP reduced the infiltration of mature dendritic cells in the liver. In vitro, TGP pretreatment ameliorated the Con A-induced mitochondrial membrane potential decline, reactive oxygen species increase, and apoptosis increase in hepatocytes. In addition, the levels of Bax, Cleaved Caspase-3 and cytoplasmic Cytochrome C decreased duringthis process, whereas those of Bcl-2 and mitochondrial Cytochrome C increased. Therefore, TGP might decrease hepatocyte apoptosis through the mitochondrial apoptotic pathway. Moreover, the maturation of bone marrow dendritic cells was also inhibited by TGP treatment. In conclusion, TGP treatment ameliorates AIH by regulating hepatocyte apoptosis and DC maturation. TGP is a potential compound for AIH treatment.

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Synergistic effects of tanshinone IIA and andrographolide on the apoptosis of cancer cells.

PMID: 

Pharmacol Rep. 2020 Feb 11. Epub 2020 Feb 11. PMID: 32048269

Abstract Title: 

Synergistic effects of tanshinone IIA and andrographolide on the apoptosis of cancer cells via crosstalk between p53 and reactive oxygen species pathways.

Abstract: 

BACKGROUND: Tanshinone IIA (Tan IIA) and andrographolide (Andro) are natural compounds that are reported to exhibit anticancer activities against various types of cancers. The aim of this study is to evaluate the synergistic anticancer effects of the combination of Tan IIA and Andro, and to investigate the mechanisms of pharmacological effect and their potential applications as an anticancer therapy in clinics.METHODS: The anticancer effects of the combination of Tan IIA and Andro on MCF7, SMMC7721, and BGC823 cells were explored. The apoptosis of the cancer cells was determined by MTT and AV-PI dual stain assays. The intracellular GSH level was measured by DTNB assay, and the intracellular levels of reactive oxygen species (ROS) were examined by flow cytometry. The expression of the proteins in the apoptosis pathway was determined by immunobloting.RESULTS: The combination of Tan IIA and Andro exhibited significant synergistic anticancer effects against cancer cells, especially at low concentrations. Andro reacted with the thiol group of intracellular GSH, thus disrupting the GSH redox cycle and eventually increasing the level of intracellular ROS. Tan IIA triggered p53 responses and apoptosis by binding to the DNA of cancer cells. The crosstalk between ROS and p53 exhibited a synergistic effect on the apoptosis of cancer cells.CONCLUSION: The combination of Tan IIA and Andro showed significant synergistic effects on cancer cell apoptosis by promoting crosstalk between ROS and p53, providing a novel and effective combination that has the potential to be applied in clinical anticancer therapy.

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