Activation Of MAPK Signaling Cascades Research Articles

Eriodictyol protects skin cells from UVA irradiation-induced photodamage by inhibition of the MAPK signaling pathway.

Solar UVA irradiation-generated reactive oxygen species (ROS) induces the expression of matrix metalloproteinase 1 (MMP-1), leading to photoaging, however the molecular mechanism remains unclear. In the present study, we found that eriodictyol remarkably reduces UVA-mediated ROS generation and protects the skin cells from oxidative damage and the ensuing cell death. Moreover eriodictyol pretreatment significantly down-regulates the UVA-induced MMP-1 expression, and lowers the inflammatory responses within the skin cells. Pretreatment with eriodictyol upregulates the expression of tissue inhibitory metalloproteinase 1 (TIMP-1) and collagen-I (COL-1) at the transcriptional level in a dose-dependent manner. UVA-induced phosphorylation levels of c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK) and p38 leading to increased MMP-1 expression are significantly reduced in eriodictyol-treated skin cells. In addition, eriodictyol pretreatment significantly suppresses inflammatory cytokines and inhibits the activation of MAPK signaling cascades in skin cells. Taken together, our results demonstrate that eriodictyol has both potent anti-inflammatory and anti-photoaging effects.

Anaerobic metabolic patterns related to stress responses in hypoxia exposed mussels Mytilus galloprovincialis

Air exposure in mussels during summer coincides with the concomitant exposure of the organism to hypoxia and ambient heat. The present study examined the metabolic and molecular stress responses of M. galloprovincialis during exposure to air at different temperatures. Mussel mortality did not rise upon 48 h of air exposure at 18° but increased significantly within the first 20 h of exposure to 32 °C. Thermal responses and limits were determined by integrating information from various levels of biological organization. Metabolic impacts were assessed by examining the activity of the key glycolytic enzyme pyruvate kinase (PK). Molecular responses were addressed through the expression of heat shock proteins Hsp70 and Hsp90 and the phosphorylation of stress-activated protein kinases, p38 mitogen-activated protein kinase (p38 MAPK) and cJun-N-terminal kinases (JNKs). PK activity decreased during exposure in a temperature dependent manner indicating a marked reduction of metabolic rate especially when M. galloprovincialis was exposed to 32 °C. The heat shock response (HSR) was stimulated in all cases of exposure indicating that hypoxia by itself may induce the expression of Hsp70 and Hsp90 in the tissues of M. galloprovincialis. Increased phosphorylation of p38 MAPK and JNKs indicate activation of MAPK signaling cascades. The potential involvement of MAPKs in the induction of Hsp genes in tissues of M. galloprovincialis during hypoxia is discussed.

Cadmium-induced induction of cell death in human lens epithelial cells: Implications to smoking associated cataractogenesis

Cadmium is reported to accumulate in human eye tissues suggesting its implication in diverse ocular pathology. Using an in vitro cell culture model we investigated the effects of cadmium on human lens epithelial cells (HLECs) (HLE-B3). We observed cadmium-induced dose- as well as time-dependent decline in HLECs viability which was exacerbated significantly upon reduction of intracellular glutathione levels by buthionine sulfoximine (BSO). There was a dose-dependent significant increase in lactate dehydrogenase (LDH) release from HLECs suggesting cadmium-induced alteration of membrane integrity as well as necrotic cell death. The decline in cell viability was also due to apoptosis of the HLECs as determined by quantifying % apoptotic cells as well as PARP cleavage. Moreover, release of apoptosis inducing factor (AIF) into the cytosol was also detected. Cadmium was also observed to increase oxidative stress, lipid peroxidation and activation of MAPK pathway in HLECs. Antioxidants like N-acetylcysteine (NAC) and α-Tocopherol significantly prevented cadmium-induced toxicity in HLECs. Our findings suggest that cadmium-induced elevated oxidative stress as well as activation of MAPK signaling cascade eventually led to cell death of HLECs through apoptosis as well as necrosis. The loss of HLECs by cadmium could possibly explain its implication in cataract development particularly associated with smoking.

Metabolic and molecular stress responses of the gilthead seabream Sparus aurata during long-term exposure to increasing temperatures

Tolerance to a changing climate regime and persistence in the natural environment depends on the limited capacity to acclimate to changing temperatures. The present study aimed to identify and characterize thermal limits of the Mediterranean fish Sparus aurata as well as the processes providing heat protection during exposure to high temperatures. Processes studied included heat shock protein expression, protein kinase activity and metabolic adjustments. Molecular responses were addressed through the expression of Hsp70 and Hsp90 and the phosphorylation of stress-activated protein kinases, p38 mitogen-activated protein kinase (p38 MAPK) and cJun-N-terminal kinases (JNKs). Thermal impacts on metabolic capacities were assessed by studying the maximum activities of citrate synthase (CS), malate dehydrogenase (MDH) and 3-hydroxyacyl CoA dehydrogenase (HOAD) as well as pyruvate kinase (PK) and lactate dehydrogenase (L-LDH). The expression of Hsp70 and hsp90 was activated when the fish were exposed to temperatures beyond 20°C. Increased phosphorylation of p38 MAPK and JNKs indicated the parallel activation of MAPK signaling cascades and the potential involvement of MAPKs in the induction of Hsp genes. Exposure to extreme temperatures beyond 24°C caused an increase in the enzymatic activity of PK and LDH indicating an enhanced glycolytic potential.

Reactive oxygen species regulate properties of transformation in UROtsa cells exposed to monomethylarsonous acid by modulating MAPK signaling

UROtsa cells exposed to 50 nM monomethylarsonous acid [MMA(III)] for 52 wk (MSC52) achieved hyperproliferation, anchorage independent growth, and enhanced tumorgenicity. MMA(III) has been shown to induce reactive oxygen species (ROS), which can lead to activation of signaling cascades causing stress-related proliferation of cells and even cellular transformation. Previous research established the acute activation of MAPK signaling cascade by ROS produced by MMA(III) as well as chronic up regulation of COX-2 and EGFR in MSC52 cells. To determine if ROS played a role in the chronic pathway perturbations by acting as secondary messengers, activation of Ras was determined in UROtsa cells [exposed to MMA(III) for 0–52 wk] and found to be increased through 52 wk most dramatically after 20 wk of exposure. Ras has been shown to cause an increase in O 2 − and be activated by increases in O 2 −, making ROS important to study in the transformation process. COX-2 upregulation in MSC52 cells was confirmed by real time RT-PCR. By utilizing both antioxidants or specific COX inhibitors, it was shown that COX-2 upregulation was dependent on ROS, specifically, O 2 −. In addition, because previous research established the importance of MAPK activation in phenotypic changes associated with transformation in MSC52 cells, it was hypothesized that ROS play a role in maintaining phenotypic characteristics of the malignant transformation of MSC52 cells. Several studies have demonstrated that cancer cells have lowered superoxide dismutase (MnSOD) activity and protein levels. Increasing levels of MnSOD have been shown to suppress the malignant phenotype of cells. SOD was added to MSC52 cells resulting in slower proliferation rates (doubling time = 42 h vs. 31 h). ROS scavengers of OH also slowed proliferation rates of MSC52 cells. To further substantiate the importance of ROS in these properties of transformation in MSC52 cells, anchorage independent growth was assessed after the addition of antioxidants, both enzymatic and non-enzymatic. Scavengers of OH, and O 2 − blocked the colony formation of MSC52 cells. These data support the role for the involvement of ROS in properties of transformation of UROtsa cells exposed to MMA(III).

Metabolic and molecular stress responses of sublittoral bearded horse musselModiolus barbatusto warming sea water: implications for vertical zonation

The present study set out to investigate the thermal limits of the Mediterranean bivalve Modiolus barbatus, acclimated to various temperatures, and includes a comparison of laboratory determined limits with its temperature-dependent restriction to deeper water layers in its natural habitat. Thermal responses and limits were determined by integrating information from various levels of biological organization, including the expression of Hsp70 and Hsp90, the phosphorylation of stress-activated protein kinases, p38 mitogen-activated protein kinase (p38 MAPK) and cJun-N-terminal kinases (JNKs) as well as metabolic adjustments. The latter were assessed by examining temperature effects on the activity of the key glycolytic enzyme pyruvate kinase (PK). The expression of Hsp70 and Hsp90 was activated when mussels were acclimated to temperatures above 20 degrees C. Increased phosphorylation of p38 MAPK and JNKs at about the same temperatures indicate activation of MAPK signaling cascades and their potential involvement in the induction of Hsp genes. As indicated by the activity of PK, Modiolus barbatus maintains some aerobic capacity when acclimated to temperatures up to 24 degrees C, while further warming probably caused metabolic depression and a shift from aerobic to anaerobic metabolism. An increase in mortality occurred in parallel, during acclimation to temperatures above 24 degrees C. Our results indicate that both the biochemical stress indicators and metabolic status respond in parallel once hypoxemia becomes extreme. Comparison with our previous study of thermal limits and vertical distribution in M. galloprovincialis dwelling in shallow waters emphasizes the relevance of maintained aerobic scope over that of passive tolerance for permanent vertical zonation at higher temperatures in the field. These findings and conclusions are in line with the concept of oxygen and capacity limited thermal tolerance and the associated systemic to molecular hierarchy of thermal limitation.

Metabolic and molecular stress responses of the gilthead seabream Sparus aurata during long term exposure to increasing temperatures

Tolerance to a changing climate regime and persistence in the natural environment depends on the limited capacity to acclimate to changing temperatures. The present study aimed to identify and characterize thermal limits of the Mediterranean fish Sparus aurata as well as the processes providing heat protection during exposure to high temperatures. Processes studied included heat shock protein expression, protein kinase activity and metabolic adjustments. Molecular responses were addressed through the expression of Hsp70 and Hsp90 and the phosphorylation of stress-activated protein kinases, p38 mitogen-activated protein kinase (p38 MAPK) and cJun-N-terminal kinases (JNKs). Thermal impacts on metabolic capacities were assessed by studying the maximum activities of citrate synthase (CS), malate dehydrogenase (MDH) and 3-hydroxyacyl CoA dehydrogenase (HOAD) as well as pyruvate kinase (PK) and lactate dehydrogenase (L-LDH). The expression of Hsp70 and hsp90 was activated when the fish were exposed to temperatures beyond 20°C. Increased phosphorylation of p38 MAPK and JNKs indicated the parallel activation of MAPK signaling cascades and the potential involvement of MAPKs in the induction of Hsp genes. Exposure to extreme temperatures beyond 24°C caused an increase in the enzymatic activity of PK and LDH indicating an enhanced glycolytic potential.

Behavioral, metabolic, and molecular stress responses of marine bivalve Mytilus galloprovincialis during long-term acclimation at increasing ambient temperature

The present study aimed to determine the thermal response of the Mediterranean mussel Mytilus galloprovincialis by integrating information from various levels of biological organization including behavior, metabolic adjustments, heat shock protein expression, and protein kinase activity. Behavioral responses were determined by examining the effect of warming on valve closure and opening. Metabolic impacts were assessed by examining the activity of the key glycolytic enzyme pyruvate kinase (PK). Molecular responses were addressed through the expression of Hsp70 and Hsp90 and the phosphorylation of stress-activated protein kinases, p38 mitogen-activated protein kinase (p38 MAPK) and cJun-N-terminal kinases (JNKs). Mussels increased the duration of valve closure by about sixfold when acclimated to 24 degrees C rather than to 17 degrees C. As indicated by the activity of PK, such behavior caused metabolic depression and probably a shift from aerobic to anaerobic metabolism. Acclimation to temperatures higher than 24 degrees C caused an increase in mortality and induced the expression of Hsp72. Increased phosphorylation of p38 MAPK and JNKs indicated activation of MAPK signaling cascades. The potential involvement of MAPKs in the induction of Hsp genes in the tissues of M. galloprovincialis is discussed. In conclusion, it seems that M. galloprovincialis lives close to its acclimation limits and incipient lethal temperature and that a small degree of warming will elicit stress responses at whole organism and molecular levels.