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Anesth Analg 2008; 107:534-540
© 2008 International Anesthesia Research Society
doi: 10.1213/ane.0b013e3181770be9
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ANESTHETIC PHARMACOLOGY

Propofol Protects Hepatic L02 Cells from Hydrogen Peroxide-Induced Apoptosis via Activation of Extracellular Signal-Regulated Kinases Pathway

Hao Wang, MD, PhD*, Zhanggang Xue, MD*, Qiong Wang, MS{dagger}, Xiaochen Feng, MS{dagger}, and Zonghou Shen, PhD{dagger}

From the *Department of Anesthesiology, Zhongshan Hospital, Fudan University, Shanghai, People’s Republic of China; and {dagger}Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, Shanghai, People’s Republic of China.

Address correspondence and reprint requests to Zhanggang Xue, MD, Department of Anesthesiology, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai 200032, People’s. Republic of China. Address e-mail to zhanggangxue{at}gmail.com.


   Abstract
Top
 Abstract
Introduction
 METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
BACKGROUND: Propofol protects cells against ischemia/reperfusion injury in several organs, but there are few reports of its effect on liver epithelial cells. We investigated the effect of propofol preconditioning on human hepatic L02 cells under hydrogen peroxide (H2O2)-induced oxidative stress and attempted to determine whether the extracellular signal-regulated kinases (ERK) pathway is involved in this process.

METHODS: Preconditioned or nonpreconditioned human hepatic L02 cells were exposed to H2O2 and the changes of apoptosis were evaluated by TUNEL assay, Caspase-3 and poly ADP-ribose polymerase (PARP) cleavage. Activation of ERK1/2 and mitogen-activated protein kinase//ERK Kinase 1/2 (MEK1/2) was measured by Western blot analysis. The mRNA expression of Bcl-2, Bcl-xL, Bad, and Bax was quantified by real-time quantitative reverse transcriptase polymerase chain reaction.

RESULTS: Propofol preconditioning reduced the population of apoptotic cells and Caspase-3 and PARP cleavage induced by H2O2 inhepatic L02 cells. L02 cells treated with propofol (0.01–0.3 mM) alone, led to a dose-dependent activation of ERK and MEK, and such activation was detected within 0.5 h and eventually declined to <50% at 4 h. The addition of the specific inhibitor PD98059 completely abolished the activation of ERK and aggravated the extent of apoptosis. Moreover, propofol treatment repressed the mRNA expression of proapoptotic genes Bad and Bax, and this repression could be partly reversed by PD98059.

CONCLUSIONS: These findings demonstrate that propofol protects hepatic L02 cells from H2O2-induced apoptosis, partly through activating the MEK-ERK pathway and further suppressing Bad and Bax expression.


   Introduction
Top
 Abstract
 Introduction
METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Liver dysfunction or failure results from ischemic-mediated cellular damage after transplantation surgery, tissue resections, and hemorrhagic shock.1,2 During these processes, intracellular generation of reactive oxygen by xanthine oxidase and, in particular, mitochondria, contribute to cell injury.3,4 The most promising protective strategy against ischemia/reperfusion (I/R) injury is preconditioning, such as ischemic preconditioning and heat shock or pharmacological interventions, which have great potential to enhance hepatocyte survival.5,6 As an IV anesthetic, propofol is used during liver transplantation because its metabolism is not greatly affected by liver failure.7 Consistent with its activity as an antioxidant, propofol has been proved to attenuate I/R injury in several organs.8–11 However, its effect on hepatocytes under oxidative stress is largely unknown.

Reactive oxygen species (ROS), including hydrogen peroxide (H2O2), are generated during I/R as a consequence of aerobic metabolism. The cellular toxicity of H2O2 is associated with the rapid modification of cellular constituents, including the depletion of intracellular glutathione and ATP, a decrease in NAD+ level, an increase in free cytosolic Ca2+, and lipid peroxidation.12 It causes cell death either by necrosis or apoptosis. Apoptosis is an active process characterized by cell shrinkage, chromatin condensation, formation of apoptotic bodies, and activation of caspases.13 Caspase-3 plays a key role in apoptotic cell death and the Poly (ADP-ribose) polymerase (PARP), a nuclear enzyme involved in DNA repair, is a well known substrate for caspase-3 cleavage. Many studies have shown that mitogen-activated protein kinase (MAPKs) activation plays a critical role in I/R cell events. Among these, extracellular signal-regulated kinases (ERK) activation has been implicated in protecting against cell injury and is considered a sign of regeneration and protection.14–16 ERK1/2 are components of a three-kinase phosphorylated module that includes MKKK c-Raf1, B-Raf, or A-Raf, which can be activated by the proto-oncogene Ras. Oncogenic Ras persistently activates ERK1/2 pathways, which contributes to the increased proliferative rate of cells. In differentiated cells, ERKs have different roles and are involved in responses.17 Via phosphorylation on Thr(202) and Tyr(204), activated ERKs interact with their substrates, including several transcription factors and other protein kinases, which in turn are responsible for the final effect of MEK-ERK signaling.18–19 Although the ERKs response has been implicated in the pathophysiology of ischemic injury, its role in propofol-engaged preconditioning in hepatic injury has not been fully identified.

In this study, we examined the effect of propofol on hepatic cell apoptosis induced by H2O2, as well as its effect on ERK1/2 activation. We designed to test the hypothesis that oxidative injury might be reversed by propofol through enhancing ERK1/2 activation in human hepatic L02 cells.


   METHODS
Top
 Abstract
 Introduction
 METHODS
RESULTS
 DISCUSSION
 REFERENCES
 
Materials
Human hepatic L02 cells are normal hepatocytes from adult liver tissue and have distinct ultrastructure compared with hepatic carcinoma cells. The cells were obtained from the Institute of Cell Biology, Academic Sinica. RPMI-1640 medium and fetal calf serum were purchased from GIBCO/BRL (Grand Island, NY). Monoclonal antibodies against human phospho-ERK1/2, {alpha}-tublin, and rabbit polyclonal antibodies against human total ERK1/2, MEK1/2, phospho-MEK1/2, Caspase-3 and PARP were the products of Cell Signaling Technology (Danvers, MA). Horseradish peroxidase (HRP)-labeled second antibodies (goat anti-rabbit IgG and anti-mouse IgG) were purchased from Dako Company. DNA in situ terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP)-biotin nick end-labeling (TUNEL) kit was from Roche (Nutley, NJ). Propofol (Diprivan, 2, 6 diisopropylphenol) was obtained from Astrazeneca Corps (Milan, Italy). Dimethyl sulfoxide (DMSO) was the product of Sigma (St. Louis, MO). ECL assay kit was from Pierce (Rockford, IL). Trizol and DNase I were from Invitrogen (Carlsbad, CA). Real-time polymerase chain reaction (PCR) regent kit ExScriptTM was purchased from TaKaRa (Dalian, China). Other regents were commercially available in China.

Cell Culture and Treatment
L02 cells were cultured at 37°C, 5% CO2 in RPMI-1640 medium containing 10% fetal calf serum, 1% penicillin (100 IU/mL), and 1% streptomycin (100 µg/mL).

Propofol dissolved in DMSO was added to the culture medium. The cells were pretreated with a final concentration of 0.01, 0.03, 0.1, 0.3 mM propofol or DMSO for 1 h, respectively. The final concentration of DMSO in all samples was <0.3% (v/v). After pretreatment with propofol, cells were stimulated with 0.2 mM H2O2 for 4–12 h to test apoptotic effects.20

To determine whether the oxidative injury caused by H2O2 or the effect of propofol were directly through the ERK pathway, L02 cells were incubated at 37°C for 1 h with 50 µM PD98059 (specific inhibitor of MEK1/2) or 0.1% DMSO, and followed by 0.3 mM propofol for 1 h before H2O2 exposure.

TUNEL Assay
Apoptosis was detected using DNA in TdT-mediated dUTP-biotin nick end-labeling (TUNEL) staining, according to the manufacturer’s protocol (Roche).21 In brief, adherent L02 cell smears and cytospin were prepared, fixed and permeated. After equilibration, cells were end labeled with digoxigenin-11-dUTP by TdT enzyme in buffer for 1 h at 37°C in a humidifying chamber. After treatment with stop-wash buffer, slides were incubated with 4'-6-diamidino-2-phenylindole (DAPI) for 30 min and observed under a fluorescence microscope. The population of apoptotic cells was counted in 10 random fields (100x).

Western Blot Analysis
In brief, cells were lysed in 1 x SDS lysis buffer (50 mM Tris-HCl pH8.0, 150 mM NaCl, 1% SDS, 100 µg/mL PMSF). Protein concentration was determined by BCA kit (Bio-color, Shanghai, China). Equal amounts of protein were separated by SDS-PAGE and transferred to NC membranes (CNI, Canada). Membranes were then blocked in TBS containing 0.1% (v/v) Tween 20 and 5% (w/v) nonfat dried milk, after which they were incubated for 2 h with the primary antibody at 1:500 in. blocking buffer, and then for 1 h with HRP-conjugated secondary antibody at 1:2000. The blots were developed using enhanced chemiluminescence and recorded by Kodak films. The intensities of the target protein bands were normalized by the intensity of {alpha}-tublin or phospho-kinase/total kinase.

Real-Time Quantitative Reverse Transcriptase
We applied quantitative real-time reverse-transcription polymerase chain reaction (RT-PCR) to quantify the expression of Bcl-2 family members, including proapoptotic genes and antiapoptotic genes. RT-PCR analysis was performed on cells treated 8 h later. Total RNA was isolated from cells by use of Trizol. First-strand cDNA was synthesized from 2 µg RNA according to the manufacturer’s protocol. Real-time quantitative RT-PCR was performed for the candidate genes and for glyceraldehydes-3-phosphate-dehydrogenase as internal control. Primer sequences specific for humans were designed using the Primer Express 2.0 software. Quantitative real-time PCR was performed in an ABI PRISM 7300 Sequencing Detector (Applied Biosystems, Foster City, CA) for SYBR green PCR master mix. Individual primers are shown in Table 1. Specificity of PCR products was verified by melting curve analysis and subsequent agarose gel electrophoresis. Each sample was analyzed in triplicate.


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  		Table 1. Gene Names and Primer Sequences Used for Real-Time Reverse-transcription Polymerase Chain Reaction

 

Statistics
All data are presented as mean ± sd. Multiple groups were compared using analysis of variance (ANOVA) followed by pairwise comparisons using Tukey’s post hoc procedure. Simple pairwise comparisons were performed using Student’s t-test where appropriate. P values <0.05 were considered statistically significant.


   RESULTS
Top
 Abstract
 Introduction
 METHODS
 RESULTS
DISCUSSION
 REFERENCES
 
H2O2 (0.2 mM) induced cell apoptosis in a time-dependent manner, whereas pretreatment with propofol in 0.01, 0.03, 0.1, or 0.3 mM gradually suppressed H2O2-induced apoptosis (Fig. 1A). Propofol preconditioning decreased the amount of cleaved caspase-3 (17 kDa) and PARP (85 kDa) (Fig. 1B). Thus, our data suggest that propofol exerts its protective role in hepatocytes cultured in vitro.


Figure 127
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  		Figure 1. Propofol protects L02 cells from H2O2-induced apoptosis. A, L02 cells were preconditioned with 0.01, 0.03, 0.1, or 0.3 mM propofol for 1 h then exposed to 0.2 mM H2O2 for 4, 8, or 12 h. Apoptotic cells were labeled by TUNEL assay. Percentage of apoptotic cells was determined by the ratio of TUNEL-positive nuclei to total nuclei number (10 fields per experiment, n = 5). *P < 0.05 versus DMSO (0 mM propofol) preconditioning. B, L02 cells were treated with different concentration of propofol then exposed to 0.2 mM H2O2 for 8 h. Expression of Caspase-3 and PAPR were determined by Western blot analysis.

 

Propofol rapidly activates ERK1/2 and MEK1/2 phosphorylation of L02 cells in a dose-dependent manner (Fig. 2A–C). Activation of ERK1/2 and MEK1/2 was detected within 0.5 h at maximum level and eventually declined to <50% at 4 h (Fig. 2D–F).


Figure 227
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  		Figure 2. Propofol activates MEK-ERK pathway in dose- and time-dependent manners. A, L02 cells were treated with indicated concentrations of propofol for 1 h, and the expression levels of pERK1/2, total EKR1/2, pMEK1/2, and total MEK1/2 were determined by Western blot analysis. B,C, The amount of pERK/ERK and pMEK/MEK was normalized to the values obtained from DMSO (0 mM propofol) treated cells. Data are presented as mean ± sd from four separated experiments. *#P < 0.05 versus DMSO (0 mM propofol) treatment. D, L02 cells were treated with 0.3 mM propofol for 0.5, 1, 2, or 4 h, and the expression levels of pERK1/2, total EKR1/2, pMEK1/2, and total MEK1/2 were determined by Western blot analysis. E-F, The amount of pERK/ERK and pMEK/MEK was normalized to the values obtained from 0 h cells. *P < 0.05 versus 0 h cells. Data are presented as mean ± sd from four separated experiments. #P < 0.05 versus 0.5 h cells.

 

H2O2 alone only slightly stimulated ERK1/2 phosphorylation in L02 cells, whereas propofol significantly induced ERK1/2 phosphorylation, and that induction was completely dependent on propofol, but not on H2O2 (Fig. 3A). PD98059 abolished almost all the phosphorylation of ERK1/2 induced by propofol (Fig. 3B). More importantly, PD98059 increased the population of apoptotic cells in H2O2-induced apoptosis when compared with control (65% ± 13% vs 30% ± 8%, P < 0.05), reversing the most protective effect of propofol (Fig. 3C and D). Results of Western blot analysis also confirmed that the amount of cleaved caspase-3 and PARP was increased after PD98059 treatment (Fig. 3I). These data suggest that ERK1/2 activation is the key effector of propofol’s antiapoptotic function.


Figure 327
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  		Figure 3. PD98059 abolishes anti-apoptotic role of propofol. A, L02 cells preconditioned with DMSO or 0.3 mM propofol for 1 h were exposed to 0.2 mM H2O2 for 1 h, and then the expression levels of pERK1/2 and total ERK1/2 were determined by Western blot analysis. The amount of pERK/ERK was normalized to the values obtained from DMSO treated cells. Data are presented as mean ± sd from four separated experiments. *#P < 0.05 versus DMSO treatment, $+P < 0.05 versus DMSO + H2O2 treatment. B, L02 cells were pretreated with 50 µM PD98059 or DMSO for 1 h before preconditioning with propofol and exposure to H2O2. Expression levels of pERK1/2 and total EKR1/2 at 1 h after H2O2 exposure were determined by Western blot analysis. The amount of pERK/ERK was normalized to the values obtained from DMSO + H2O2 treated cells. Data are presented as mean ± sd from four separated experiments. *#P < 0.05 versus DMSO + H2O2 treatment. $+P < 0.05 versus Propofol + H2O2 treated cells. C-G, Cell apoptosis at 8 h after H2O2 exposure was measured by TUNEL assay. C: DMSO, D: DMSO + H2O2, E: Propofol + H2O2, F: PD98059 + Propofol+H2O2, G: PD98059 + H2O2. Bar: 200 µm. H. Percentage of apoptotic cells in the presence of DMSO or PD98059 was determined by the ratio of TUNEL-positive nuclei to total nuclei number (10 fields per experiment, n = 5). *P< 0.05 versus Propofol + H2O2 treatment. I. Western blot analysis of Caspase-3 and PARP of apoptotic cells. The amount of cleaved Caspase-3 and PARP was normalized to the values obtained from DMSO + H2O2 treated cells. *P < 0.05 versus Propofol + H2O2 treatment.

 

Real-time RT-PCR results showed that propofol alone repressed the mRNA expression of Bad and Bax, two proapoptotic genes, whereas the expression of Bcl-xLand Bcl-2, two anti-apoptotic genes, had no significant alteration (Fig. 4A). During H2O2-induced apoptosis, Bad, Bax, Bcl-xL, and Bcl-2 were all moderately repressed. The mRNA levels of Bad and Bax were further reduced under propofol preconditioning (Fig. 4B). PD98059 partly abolished this repression (Fig. 4C), indicating that the Bad and Bax expression was downregulated by propofol in an ERK1/2-dependent manner.


Figure 427
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  		Figure 4. Propofol regulates the mRNA expression of Bad and Bax. A, L02 cells were treated with 0.3 mM propofol or DMSO for 8 hours, and then the relative mRNA levels of Bcl-2, Bcl-xL, Bad, and Bax genes were detected by quantitative RT-PCR. Data are presented as mean ± sd from four separated experiments. *P < 0.05 versus control (DMSO treatment). B, L02 cells were preconditioned with DMSO or propofol for 1 h before exposure to H2O2 for 8 hours. Relative mRNA levels of Bcl-2, Bcl-xL, Bad, and Bax genes were detected by quantitative RT-PCR. Data are presented as mean ± sd from four separated experiments. *P < 0.05 versus control (DMSO treatment). #P < 0.05 versus DMSO + H2O2 treatment. C, L02 cell were treated with PD98059 or DMSO before propofol preconditioning and exposure to H2O2. Relative mRNA levels of Bcl-2, Bcl-xL, Bad, and Bax genes at 8 h after H2O2 exposure were detected by quantitative RT-PCR. Data are presented as mean ± sd from four separated experiments. *P < 0.05 versus control (DMSO treatment). #P < 0.05 versus DMSO + Propofol + H2O2 treatment.

 


   DISCUSSION
Top
 Abstract
 Introduction
 METHODS
 RESULTS
 DISCUSSION
REFERENCES
 
The purpose of this study was to elucidate the role of propofol in hepatocytes under oxidative stress. All data here provide strong evidence that propofol acts as a protective modifier of apoptosis in hepatic L02 cells, and this effect is partly brought about through activation of the MEK-ERK pathway and further repression of particular proapoptotic gene expression.

To investigate the role of propofol during oxidant-mediated apoptosis, we used H2O2 as the apoptosis inducer. According to our experiments, H2O2 caused apparent apoptosis in L02 cells after 8 h exposure. To assess the effects of propofol accurately, it is important to exclude the interference of the solvent. Some reports have implied that intralipid affected neutrophil activity in a concentration-dependent manner and inflicted significant damage on the perfused murine liver.22 Therefore, we used propofol dissolved in DMSO to exclude the potential impact of emulsion. In our study, the solvent DMSO alone (<0.3%) did not affect the apoptosis induced by H2O2, whereas propofol preconditioning exerted protective role in a dose-dependent manner. At the clinically relevant concentration 0.03 mM, propofol started to decrease the population of apoptotic cells and the amount of cleaved caspase-3 and PARP, and more efficient protection was observed at 0.1 and 0.3 mM. Our study shows that propofol protects hepatocytes in vitro, suggesting its potential benefit in liver surgery.

We also have shown that propofol, at clinically achievable concentrations (≥0.01 mM), activated ERK1/2 phosphorylation in hepatocytes. Although some researchers have reported the relationship of ERK activation with propofol, this subject remains controversial. Nagata et al. found that propofol inhibits ERK activation in human neutrophils.23 Kozinn et al. found that propofol eliminated the glutamatergic activation of the ERK pathway in hippocampal neurons.24 In our experiment, 0.2 mM H2O2 alone had no obvious effect on ERK activation in hepatic L02 cells, unlike in cardiac myocytes, where ERK could be activated by H2O2. Interestingly, propofol activates the ERK pathway in both dose- and time-dependent manners. Furthermore, PD98059 markedly inhibits propofol-induced ERK activation and partly reverses the protective role of propofol towards H2O2-induced apoptosis. These data suggest an essential role of ERK activation for propofol activity, but also indicate that other signaling pathway might be involved in the protective effect of propofol.

The Bcl-2 family of proteins function as key regulators of apoptosis. It consists of two classes: anti-apoptotic genes, such as Bcl-xL and Bcl-2, proven to promote survival, and pro-apoptotic genes including Bad and Bax, which oppose survival function. Indeed, the ratio between these two subsets determines the susceptibility of cells to a death signal.25,26 In clinical investigations, some antioxidative drugs, such as manganese superoxide dismutase (MnSOD), regulate the ratio of Bax and Bcl-2. Based on previous reports that both ROS and MAPKs regulate the expression of Bcl-2, Bcl-xL, Bad, and Bax, we examined whether propofol regulates the expression of these genes. Interestingly, after propofol treatment, mRNA expression of Bcl-2 and Bcl-xL did not change, but Bad and Bax were greatly downregulated. Consequently, the ratio between anti- and proapoptotic genes was considerably increased. The changes of these molecules further explained why propofol promoted L02 cell survival. Although some studies showed that increased expression of Bcl-2 confers cellular resistance to apoptosis under various stresses,27 we did not observe obvious changes in Bcl-2 expression upon propofol treatment. The changes of these molecules further explained why propofol promoted L02 cell survival. It has been reported that multiple protein kinases, including MAPKs, may phosporylate Bcl-2 and stimulate its antiapoptotic function.28 A recent study found that propofol reduced tumor necrosis factor induced apopotosis by effecting Bcl-2 and Bax expression.29 But Oshiro et al. reported that overexpression of Bcl-2 promotes apoptosis in hepatocyte during I/R injury.30 In our experiments, we observed not upregulation but, rather, a slight reduction of Bcl-xL by propofol. These data implied that supression of Bcl-xL by PD 98059 might be a secondary, but not a direct, effect of ERK inhibition.

Considering that all data came from the in vitro experiments, our results are not in agreement with some animal and clinical experiments.31 Considering the limited number of reports, the effect of propofol on the liver during I/R injury remains controversial. Navapurkar et al.9 reported an antiapoptotic role of propofol in rat hepatocytes under oxidant stress, but Shimono et al. found that propofol had no effect on hypoxia injury in rat liver slices.32,33 The reasons the discrepancy might be as following: 1) L02 cells are uniform hepatocytes derived from a single clone, excluding the other cell types, such as kupper cells and neutrophils which produce ROS and counteract antioxidative effects, 2) we used a wide range of propofol concentrations exceeding the clinically relevant concentration to test the effect; 3) Since propofol is highly lipid soluble, it is likely to react in lipophilic environments. The in vitro environment cannot completely imitate in vivo conditions, which may mask the role of propofol.

Because our data showed that propofol activates MEK/ERK rapidly, we proposed that propofol should directly activate certain upstream activators of the MEK/ERK pathway, such as receptors on cellular membrane, or intracellular signaling molecules. Consistent with our hypothesis, Oscarsson et al. reported recently that the {gamma}-aminobutyric acid receptor regulates ERK activation induced by propofol in neurones.34 Whatever the ERK pathway involved in propofol’s protective role, our study does not exclude the possible involvement of other kinases, including PI-3K.35 Acquaviva et al. suggested that propofol utilizes heme oxygenase (HO)-1 in exerting its antiapoptotic effect and this pathway has not yet been examined in hepatocytes.8 Based on our present results, further examination is required to determine whether propofol activates the MEK-ERK pathway in other, different I/R models, such as hyperosmotic stress, anoxia/reoxygenation or mechanical overload.36–38 In addition, it is still unknown whether the ERK affects the phosphorylation of Bcl-2 family proteins after propofol treatment, which also determines cell fate.

In conclusion, our results suggest that the protective effect of propofol on hepatocytes is partly mediated by activating the ERK-MAPK pathway and downregulating the mRNA expression of Bax and Bad.


   Footnotes
 
Accepted for publication March 20, 2008.


   REFERENCES
Top
 Abstract
 Introduction
 METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 

  1. Jaeschke H. Molecular mechanisms of hepatic ischemia-reperfusion injury and preconditioning. AJP-Gastrointest Liver Physiol 2003;284:15–26
  2. Li CY, Jcakson RM. Reactive species mechanisms of cellular hypoxia-reoxygenation injury. Am J Physiol Cell Physiol 2002;282:227–41
  3. Petrosillo G, Ruggiero FM, Pistolese M, Paradies G. Ca2+-induced reactive oxygen species production promotes cytochrome c release from rat liver mitochondria via mitochondrial permeability transition (MPT)-dependent and MPT-independent mechanisms: role of cardiolipin. J Biol Chem 2004;279:53103–8[Abstract/Free Full Text]
  4. Zhong Z, Lemasters JJ, Thurman RG. Role of purines and xanthine oxidase in reperfusion injury in perfused rat liver. J Pharmacol Exp Ther 1989;250:470–75[Abstract/Free Full Text]
  5. Pasupathy S, Homer-Vanniasinkam S. Surgical implications of ischemic preconditioning. Arch Surg 2005;140:405–9[Abstract/Free Full Text]
  6. Zaugg M, Lucchinetti E, Uecker M, Pasch T, Schaub MC. Anaesthetics and cardiac preconditioning. Part I. Signalling and cytoprotective mechanisms. Br J Anaesth 2003;91:551–65[Abstract/Free Full Text]
  7. Takizawa D, Sato E, Hiraoka H, Tomioka A, Yamamoto K, Horiuchi R, Goto F. Changes in apparent systemic clearance of propofol during transplantation of living related donor liver. Br J Anaesth 2005;95:643–7[Abstract/Free Full Text]
  8. Acquaviva R, Campisi A, Murabito P, Giuseppina R, Avola R, Mangiameli S, Musumeci L, Barcellona ML, Vanella A, Volti GL. Propofol attenuates peroxynitrite-mediated DNA damage and apoptosis in cultured astrocytes. Anesthesiology 2004;101: 363–71
  9. Navapurkar VU, Skepper JN, Jones JG, Menon DK. Propofol preserves the viability of isolated rat hepatocyte suspensions under an oxidant stress. Anesth Analg 1998;87:1152–57[Abstract/Free Full Text]
  10. Englelhard K, Werner C, Eberspacher E, Pape M, Blobner M, Hutzler P, Kochs E. Sevoflurane and propofol influence the expression of apoptosis-regulating proteins after cerebral ischaemia and reperfusion in rats. Eur J Anaesthesiol 2004;21: 530–37[Web of Science][Medline]
  11. Luo T, Xia ZY, Ansey DM, Ouyang JP, Granville DJ, Li YP, Xia ZY, Zhou QS, Liu ZY. Propofol dose-dependently reduced tumor Necrosis factor-alpha-induced human umbilical vein endothelial cell apoptosis: effects on Bcl-2 and Bax expression and nitric oxide generation. Anesth Analg 2005;100:1653–9[Abstract/Free Full Text]
  12. Navarro A, Boveris A. The mitochondrial energy transduction system and the aging process. Am J Physiol Cell Physiol 2007;292:670–86
  13. Jones BA, Gores GJ. Physiology and pathophysiology of apoptosis in epithelial cells of the liver, pancreas, and intestine. Am J Physiol Gastrointest Liver Physiol 1997;273:1174–88
  14. Cavanaugh JE, Jaumotte JD, Lakoski JM, Zigmond MJ. Neuroprotective role of ERK1/2 and ERK5 in a dopaminergic cell line under basal conditions and in response to oxidative stress. J Neurosci Res 2006;84:1367–75[Web of Science][Medline]
  15. Nair VD, Yuen T, Olanow CW, Sealfon SC. Early single cell bifurcation of pro- and antiapoptotic states during oxidative stress. J Biol Chem 2004;279:27494–501[Abstract/Free Full Text]
  16. Xia ZG, Dickens M, Raingeaud J, Davis RJ, Greenberg ME. Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis. Science 1995;270:1326–31[Abstract/Free Full Text]
  17. Johnson GL, Lapadat R. Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases. Science 2002 6;298:1911
  18. Di Mari JF, Davis R, Safirstein RL. MAPK activation determines renal epithelial cell survival during oxidative injury. Am J Physiol 1999;277:195–203
  19. Gaitanaki C, Konstantina S, Chrysa S, Beis I. Oxidative stress stimulates multiple MAPK signaling pathways and phosphorylation of the small HSP27 in the perfused amphibian heart. J Exp Biol 2003;206:2759–69[Abstract/Free Full Text]
  20. Hermann C, Zeiher AM, Dimmeler S. Shear stress inhibits H2O2-induced apoptosis of human endothelial cells by modulation of the glutathione redox cycle and nitric oxide synthase. Arterioscler Thromb Vasc Biol 1997;17:3588–92[Abstract/Free Full Text]
  21. Gavrieli Y, Sherman Y, Ben-Sasson SA. Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol 1992;119:493–501[Abstract/Free Full Text]
  22. Baker MT, Gregerson MS, Naguib M. Role of lipid in sulfite-dependent propofol dimerization. Anesthesiology 2004;100: 1235–41[Web of Science][Medline]
  23. Nagata T, Kansha M, Irita K, Takahashi S. Propofol inhibits FMLP-stimulated phosphorylation of p42 mitogen-activated protein kinase and chemotaxis in human neutrophils. Br J Anaesth 2001;86:853–58[Abstract/Free Full Text]
  24. Kozinn J, Mao L, Arora A, Yang L, Fibuch EE, Wang JQ. Inhibition of glutamatergic activation of extracellular signal-regulated protein kinases in hippocampal neurons by the intravenous anesthetic propofol. Anesthesiology 2006;105:1182–91[Web of Science][Medline]
  25. Pantano C, Shrivastava P, McElhinney B, Janssen-Heininger Y. Hydrogen peroxide signaling through tumor necrosis factor receptor 1 leads to selective activation of c-Jun N-terminal kinase. J Biol Chem 2003;278:44091–96[Abstract/Free Full Text]
  26. Adams JM, Cory S. The Bcl-2 protein family: arbiters of cell survival. Science 1998;281:1322–26[Abstract/Free Full Text]
  27. Deng XM, Gao FQ, May WS. Bcl2 retards G1/S cell cycle transition by regulating intracellular ROS. Blood 2003;102: 3179–85[Abstract/Free Full Text]
  28. Hockenbery DM, Oltvai ZN, Yin XM, Milliman CL, Korsmeyer SJ. Bcl-2 functions in an antioxidant pathway to prevent apoptosis. Cell 1993;75:241–51[Web of Science][Medline]
  29. Luo T, Xia ZY. A small dose of hydrogen peroxide enhances tumor necrosis factor-alpha toxicity in inducing human vascular endothelial cell apoptosis: reversal with propofol. Anesth Analg 2006;103:110–6[Abstract/Free Full Text]
  30. Oshiro T, Shiraishi M, Muto Y. Adenovirus mediated gene transfer of antiapoptotic protein in hepatic ischemia-reperfusion injury: the paradoxical effect of Bcl-2 expression in the reperfused liver. J Surg Res 2002;103:30–6[Web of Science][Medline]
  31. Tsuchiya M, Asada A, Arita K, Utsumi T, Yoshida T, Sato EF, Utsumi K, Inoue M. Induction and mechanism of apoptotic cell death by propofol in HL-60 cells. Acta Anaesthesiol Scand 2002;46:1068–74[Web of Science][Medline]
  32. Anand K, Ramsay MA, Crippin JS. Hepatocellular injury following the administration of propofol. Anesthesiology 2001 95:1523–4[Web of Science][Medline]
  33. Shimono H, Goromaru T, Kadota Y, Tsurumaru T, Kanmura Y. Propofol displays no protective effect against hypoxia/ reoxygenation injury in rat liver slices. Anesth Analg 2003;97: 442–8[Abstract/Free Full Text]
  34. Oscarsson A, Juhas M, Sjolander A, Eintrei C. The effect of propofol on actin, ERK-1/2 and GABA(A) receptor content in neurones. Acta Anaesthesiol Scand 2007;51:1184–89[Web of Science][Medline]
  35. Datta SR, Brunet A, Greenberg ME. Cellular survival: a play in three Akts. Genes Dev 1999;13:2905–27[Free Full Text]
  36. Fayard E, Tintignac LA, Baudry A, Hemmings BA. Protein kinase B/Akt at a glance. J Cell Sci 2005;118:5675–78[Free Full Text]
  37. Bickler PE. Clinical perspectives: neuroprotection lessons from hypoxia-tolerant organisms. J Exp Biol 2004;207:3243–49[Abstract/Free Full Text]
  38. Bell LM, Leong ML, Kim B, Wang E, Park J, Hemmings BA, Firestone GL. Hyperosmotic stress stimulates promoter activity and regulates cellular utilization of the serum- and glucocorticoid-inducible protein kinase (Sgk) by a p38 MAPK-dependent pathway. J Biol Chem 2000;275:25262–272[Abstract/Free Full Text]




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