scholarly journals Marine Cyclic Dipeptide Cyclo (L-Leu-L-Pro) Protects Normal Breast Epithelial Cells from tBHP-induced Oxidative Damage by Targeting CD151

2021 ◽  
pp. 162-173
Author(s):  
Deepak KGK ◽  
Seema Kumari ◽  
Rama Rao Malla
Keyword(s):  
Oxidative Stress ◽  
Hydrogen Peroxide ◽  
Gene Silencing ◽  
Cyclic Peptide ◽  
Normal Breast ◽  
Cellular Damage ◽  
Cyclic Dipeptide ◽  
Protective Activity ◽  
Intracellular Ros ◽  
Breast Epithelial

Background: Oxidative stress plays a key role in breast carcinogenesis. Cyclo (L-Leu-L-Pro) (CLP) is a homodetic cyclic dipeptide with 2,5-diketopiperazine scaffold isolated from marine actinobacteria. This study aimed to evaluate the protective activity of CLP and linear - (L-Leu-L-Pro) (LP) from tert-butyl hydroperoxide (tBHP)-induced damage using normal breast epithelial cell line model (MCF-12A). Methods: The cytoprotective activity was evaluated by detecting the changes in intracellular ROS, mitochondrial superoxide, hydroxyl radical, hydrogen peroxide, and lipid peroxidation detection assays as well as cytotoxic assays of MTT, LDH assays and phase contrast microscopy. Genoprotective activity was evaluated by (Apurinic/Apyrimidinic) AP site, alkaline Comet, and 8-hydroxy-2-deoxyguanosine assays. Results: The marine cyclic peptide, CLP, significantly protected MCF-12A cells by scavenging tBHP induced intracellular ROS such as super oxide, hydroxyl radicals and hydrogen peroxide, and by reducing the cytotoxicity and genotoxicity effect compared to LP. Moreover, the results showed that CD151 gene silencing by shRNA significantly reduced the overexpression of CD151, tBHP-induced ROS generation, cytotoxicity and genotoxicity in MCF-12A cells. The overexpression of CD151 caused increased levels of cytochrome P450, but was reduced following the application of CD151shRNA and CLP which led to elevated levels of intracellular ROS. Conclusion: In the present study we noticed that CD151 gene silencing by shRNA and treatment with CLP have similar effects on reducing the intracellular ROS. This study uncovers the protective activity of CLP against a CD151-mediated oxidative stress-induced cellular damage. Our observations suggest that the anti-stress and anti-inflammation properties of CLP might have implications in cancer and are worth testing in cancer cell lines and tumor cells.

Dynamic Mitochondrial Responses to Oxidative Stress in Primary AML Cells Involve Structural Damage and Repair Activity.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4139-4139
Author(s):  
Myung-Geun Shin ◽  
Hye-Ran Kim ◽  
Hyeoung-Joon Kim ◽  
Mi-Ji Kim ◽  
Kook Hoon ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Hydrogen Peroxide ◽  
Mrna Expression ◽  
Blood Cells ◽  
Normal Blood ◽  
Mtdna Damage ◽  
Mtdna Mutations ◽  
Intracellular Ros ◽  
Repair Activity ◽  
Mtdna Repair

Abstract Mitochondrial DNA (mtDNA) is particularly susceptible to oxidative damage and mutations because of the high level of reactive oxygen species (ROS) generated and the inefficiency of the mtDNA repair system. The oxidative stress elicited by chronic inflammation increases the number of mtDNA mutations and might correlate with a cancerous status. We postulated that increased oxidative stress in primary AML cells might cause mtDNA damage, which can lead to mtDNA mutations, structural changes, perturbation of mtDNA repair and biogenesis. Many mutation and polymorphisms (a total of 606 mtDNA sequence variants) were identified from 48 matched AML bone marrow and buccal mucosa samples, and blood samples from 57 control subjects. There were profound alterations in the 303 poly C, 16184 poly C, and 514 CA repeats. The intracellular ROS generation of cells can be investigated using the 2′,7′-dichlorfluorescein-diacetate and flow cytometry. The results were expressed as mean fluorescence intensity (MFI). MFI in primary AML cells (4,435±709) was significantly higher than those in control blood cells (1,562±141) (P<0.05). After then, we checked mRNA expression of peroxisome proliferators activated receptor gamma coactivator-1α (PGC-1α), PGC-1-related coactivator (PRC) and nuclear respiratory factor 1 (NRF-1) because these have been identified as an important regulator of intracellular ROS level and crucial factors linking external stimuli to mitochondrial biogenesis. As compared to normal blood cells, about 2.0 fold increase in NRF-1 mRNA expression was noted in primary AML, whereas PGC-1α and PRC mRNA expression were not remarkably changed. The supercoiled and open relaxed forms of mtDNA reflect functional and damaged molecules. Thus, sensitive detection of the relaxed and total mtDNA from the same DNA templates should allow quantitative measurements on mtDNA damage, repair and copy number change in stressed cells (Nucleic Acids Res2007;35:1377–88). Primary AML cells and normal blood cells were exposed to sublethal concentration of hydrogen peroxide to study mtDNA damage and repair activity using real-time PCR quantification. When AML cells and normal blood cells were treated with 240uM hydrogen peroxide, an average of 2.1 and 2.2 fold decreases in mtDNA amplification of AML cells were detected in two mtDNA markers (HV2 and cytochrome b) after 1-h and 2-h exposure, respectively. The exposure time-dependent increase in mtDNA amplification due to increased proportion of open relaxed forms was observed in normal blood cells after 15 min to 2-h treatment of hydrogen peroxide. In conclusion, the high level of intracellular ROS in primary AML cells might cause mtDNA damage, which can lead to mtDNA mutations and increased mitochondrial biogenesis by NRF-1 activation. Although the limited repair capacity hypothesis has been validated experimentally in some experimental systems, this study showed elevated mtDNA repair activity compared to normal blood cells. Our data also support the possibility that NRF-1 targeting approach may aid in the treatment of AML.

scholarly journals Hydrogen Peroxide Causes Cell Death via Increased Transcription of HOXB13 in Human Lung Epithelial A549 Cells

Toxics ◽  
2020 ◽  
Vol 8 (4) ◽  
pp. 78
Author(s):  
Naoki Endo ◽  
Takashi Toyama ◽  
Akira Naganuma ◽  
Yoshiro Saito ◽  
Gi-Wook Hwang
Keyword(s):  
Oxidative Stress ◽  
Hydrogen Peroxide ◽  
Cell Death ◽  
Human Lung ◽  
A549 Cells ◽  
Protein Levels ◽  
Intracellular Ros ◽  
Lung Epithelial ◽  
Homeobox Protein

Although homeobox protein B13 (HOXB13) is an oncogenic transcription factor, its role in stress response has rarely been examined. We previously reported that knockdown of HOXB13 reduces the cytotoxicity caused by various oxidative stress inducers. Here, we studied the role of HOXB13 in cytotoxicity caused by hydrogen peroxide in human lung epithelial A549 cells. The knockdown of HOXB13 reduced hydrogen peroxide-induced cytotoxicity; however, this phenomenon was largely absent in the presence of antioxidants (Trolox or N-acetyl cysteine (NAC)). This suggests that HOXB13 may be involved in the cytotoxicity caused by hydrogen peroxide via the production of reactive oxygen species (ROS). Hydrogen peroxide also increased both the mRNA and protein levels of HOXB13. However, these increases were rarely observed in the presence of a transcriptional inhibitor, which suggests that hydrogen peroxide increases protein levels via increased transcription of HOXB13. Furthermore, cell death occurred in A549 cells that highly expressed HOXB13. However, this cell death was mostly inhibited by treatment with antioxidants. Taken together, our findings indicate that HOXB13 may be a novel factor involved in the induction of oxidative stress, which causes cell death via intracellular ROS production.

Integrin β3 prevents apoptosis of HL-1 cardiomyocytes under conditions of oxidative stressThis article is one of a selection of papers published in a Special Issue on Oxidative Stress in Health and Disease.

2010 ◽  
Vol 88 (3) ◽  
pp. 324-330 ◽  
Author(s):  
Lee J. Shewchuk ◽  
Sean Bryan ◽  
Marina Ulanova ◽  
Neelam Khaper
Keyword(s):  
Oxidative Stress ◽  
Hydrogen Peroxide ◽  
Flow Cytometry ◽  
Gene Silencing ◽  
Cardiac Remodeling ◽  
Caspase 3 ◽  
Annexin V ◽  
Fold Increase ◽  
Cardiomyocyte Apoptosis ◽  
Active Caspase

Integrin receptors are essential in the regulation of vital cardiac functions, and impaired integrin activity has been associated with cardiac remodeling. Oxidative stress is known to be involved in apoptosis and cardiac remodeling and thus may profoundly influence cardiac function via integrin modulation. The aim of this study was to determine the expression pattern and functional role of integrins in HL-1 cardiomyocytes under conditions of oxidative stress. Gene expression was studied by end-point and real-time PCR; surface protein expression was studied by flow cytometry; integrin knockdown was accomplished by siRNA gene silencing; and apoptosis was studied by annexin V staining and active caspase-3/7 using flow cytometry. Among the various subunits under study (αv, α5, α6, and β1, β3, β4, and β5), the expression of β3 integrin was significantly increased at both the mRNA and protein levels in cardiomyocytes exposed to 100 µmol/L hydrogen peroxide for 3 h. Gene silencing of β3 integrin by using siRNA resulted in a 2-fold increase in cardiomyocyte apoptosis upon treatment with hydrogen peroxide. This increase in apoptosis, as measured by annexin V staining, correlated with an increase in active caspase-3/7. Integrin β3 plays a vital role in preventing cardiomyocyte apoptosis under conditions of oxidative stress.

scholarly journals Modulation of Hydrogen Peroxide-Induced Oxidative Stress in Human Neuronal Cells by Thymoquinone-Rich Fraction and Thymoquinone via Transcriptomic Regulation of Antioxidant and Apoptotic Signaling Genes

2016 ◽  
Vol 2016 ◽  
pp. 1-15 ◽  
Author(s):  
Norsharina Ismail ◽  
Maznah Ismail ◽  
Nur Hanisah Azmi ◽  
Muhammad Firdaus Abu Bakar ◽  
Hamidon Basri ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Hydrogen Peroxide ◽  
Nigella Sativa ◽  
Morphological Observation ◽  
Cell Viability Assay ◽  
Viability Assay ◽  
Expression Of Genes ◽  
Intracellular Ros ◽  
Human Neuronal Cells ◽  
Better Than

Nigella sativaLinn. (N. sativa) and its bioactive constituent Thymoquinone (TQ) have demonstrated numerous pharmacological attributes. In the present study, the neuroprotective properties of Thymoquinone-rich fraction (TQRF) and TQ against hydrogen peroxide- (H2O2-) induced neurotoxicity in differentiated human SH-SY5Y cells were investigated. TQRF was extracted using supercritical fluid extraction while TQ was acquired commercially, and their effects on H2O2were evaluated using cell viability assay, reactive oxygen species (ROS) assay, morphological observation, and multiplex gene expression. Both TQRF and TQ protected the cells against H2O2by preserving the mitochondrial metabolic enzymes, reducing intracellular ROS levels, preserving morphological architecture, and modulating the expression of genes related to antioxidants (SOD1, SOD2, and catalase) and signaling genes (p53, AKT1, ERK1/2, p38 MAPK, JNK, and NF-κβ). In conclusion, the enhanced efficacy of TQRF over TQ was likely due to the synergism of multiple constituents in TQRF. The efficacy of TQRF was better than that of TQ alone when equal concentrations of TQ in TQRF were compared. In addition, TQRF also showed comparable effects to TQ when the same concentrations were tested. These findings provide further support for the use of TQRF as an alternative to combat oxidative stress insults in neurodegenerative diseases.

scholarly journals Vitamin E Can Ameliorate Oxidative Damage of Ovine Hepatocytes In Vitro by Regulating Genes Expression Associated with Apoptosis and Pyroptosis, but Not Ferroptosis

Molecules ◽  
2021 ◽  
Vol 26 (15) ◽  
pp. 4520
Author(s):  
Luyang Jian ◽  
Ying Xue ◽  
Yuefeng Gao ◽  
Bo Wang ◽  
Yanghua Qu ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Cell Death ◽  
Vitamin E ◽  
Oxidative Damage ◽  
Treated Group ◽  
Cellular Damage ◽  
Control Group ◽  
Group 4 ◽  
Intracellular Ros

(1) Background: the current research was conducted to investigate the potential non-antioxidant roles of vitamin E in the protection of hepatocysts from oxidative damage. (2) Methods: primary sheep hepatocytes were cultured and exposed to 200, 400, 600, or 800 μmol/L hydrogen peroxide, while their viability was assessed using a CCK-8 kit. Then, cells were treated with 400 μmol/L hydrogen peroxide following a pretreatment with 50, 100, 200, 400, and 800 μmol/L vitamin E and their intracellular ROS levels were determined by means of the DCF-DA assay. RNA-seq, verified by qRT-PCR, was conducted thereafter: non-treated control (C1); cells treated with 400 μmol/L hydrogen peroxide (C2); and C2 plus a pretreatment with 100 μmol/L vitamin E (T1). (3) Results: the 200–800 μmol/L hydrogen peroxide caused significant cell death, while 50, 100, and 200 μmol/L vitamin E pretreatment significantly improved the survival rate of hepatocytes. ROS content in the cells pretreated with vitamin E was significantly lower than that in the control group and hydrogen-peroxide-treated group, especially in those pretreated with 100 μmol/L vitamin E. The differentially expressed genes (DEGs) concerning cell death involved in apoptosis (RIPK1, TLR7, CASP8, and CASP8AP2), pyroptosis (NLRP3, IL-1β, and IRAK2), and ferroptosis (TFRC and PTGS2). The abundances of IL-1β, IRAK2, NLRP3, CASP8, CASP8AP2, RIPK1, and TLR7 were significantly increased in the C1 group and decreased in T1 group, while TFRC and PTGS2 were increased in T1 group. (4) Conclusions: oxidative stress induced by hydrogen peroxide caused cellular damage and death in sheep hepatocytes. Pretreatment with vitamin E effectively reduced intracellular ROS levels and protected the hepatocytes from cell death by regulating gene expression associated with apoptosis (RIPK1, TLR7, CASP8, and CASP8AP2) and pyroptosis (NLRP3, IL-1β, and IRAK2), but not ferroptosis (TFRC and PTGS2).

Effect of Turmeric and Ginger on Lipid Profile in Male Rats Exposed to Oxidative Stress

2019 ◽  
Vol 10 (01) ◽  
Author(s):  
Eman A. Al-Rekabi ◽  
Dheyaa K. Alomer ◽  
Rana Talib Al-Muswie ◽  
Khalid G. Al-Fartosi
Keyword(s):  
Oxidative Stress ◽  
Hydrogen Peroxide ◽  
Drinking Water ◽  
Lipid Profile ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Male Rats ◽  
Control Group ◽  
Very Low Density Lipoprotein ◽  
Low Density

The present study aimed to investigate the effect of turmeric and ginger on lipid profile of male rats exposed to oxidative stress induced by hydrogen peroxide H2O2 at a concentration of 1% given with consumed drinking water to male rats. Methods: 200 mg/kg from turmeric and ginger were used, and the animals were treatment for 30 days. Results: the results showed a significant increase in cholesterol, triglycerides, low density lipoprotein (LDL), very low density lipoprotein (VLDL), whereas it explained a significant decrease in high density lipoprotein (HDL) of male rats exposed to oxidative stress when compared with control group. the results showed a significant decrease in cholesterol, triglycerides, (LDL), (VLDL), whereas it explained a significant increase in (HDL) of rats treated with turmeric and ginger at dose 200 mg/kg when compared with male rats exposed to oxidative stress.

Antioxidative Effect of Rhus javanica Linne Extract Against Hydrogen Peroxide or Menadione Induced Oxidative Stress and DNA Damage in HepG2Cells

2004 ◽  
Vol 9 (2) ◽  
pp. 150-155 ◽  
Author(s):  
Chi-Sung Chun ◽  
Ji-Hyun Kim ◽  
Hyun-Ae Lim ◽  
Ho-Yong Sohn ◽  
Kun-Ho Son ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Hydrogen Peroxide ◽  
Dna Damage ◽  
Antioxidative Effect
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