Curcumin Treatment Protects Against Renal Ischemia and Reperfusion InjuryeInduced Cardiac Dysfunction and Myocardial Injury T.-H. Chen, Y.-C. Yang, J.-C. Wang, and J.-J. Wang ABSTRACT Objectives. Renal ischemia and reperfusion (I/R) injury frequently leads to acute renal failure (ARF) and multiple-organ injury with a substantial morbidity rate. The primary cause of ARF-associated death is, however, cardiac failure instead of renal failure itself, and the pathogenesis of renal I/R-induced cardiac injury is still poorly understood. We evaluated the efficacy of curcumin pretreatment on cardioprotection. Methods. Thirty Sprague-Dawley rats were evenly divided into 3 groups of shamoperated control, renal I/R injury, and a curcumin pretreatment group. Renal ischemia was conducted by bilateral occlusions of pedicles for 45 minutes, followed by 3 hours of reperfusion. The cardiac function was assessed by the left ventricular end-systolicpressure-volume-relation (ESPVR), systolic pressure (SP), ejection fraction (EF), and stroke volume (SV). Myocardial injury was assessed based on creatine kinase muscle brain fraction (CK-MB) and Troponin I (cTnI), and kidney injury was assessed based on blood urea nitrogen (BUN) and creatinine. We also assessed the levels of tumor necrosis factor-a (TNF-a) and malondialdehyde (MDA) in the heart tissues. Results. SV, EF, and SP reduced moderately during the ischemic phase with no major change in ESPVR. During reperfusion, SV, SP, and ESPVR initially increased, and then steadily decreased. Myocardial and kidney injury were marked by the increases in serum CK-MB and cTnI, and creatinine and BUN level. Curcumin pretreatment ameliorated ESPVR and attenuated injuries of both the heart and kidney resulting from I/R insult. Conclusions. Curcumin pretreatment improved cardiac contractility and attenuated myocardial and renal injury through reducing inflammatory response in the kidney and heart and oxidative stress in the myocardium.
R
ENAL ischemia and reperfusion (I/R) injury frequently occurred following sepsis, vasoconstrictive drugs, and kidney transplantation, which is the major cause of acute renal failure (ARF) resulting in significant morbidity and mortality.1 However, cardiac failure, instead of renal failure itself, has been observed as the primary cause of death in this clinical condition. Myocardial depression factors, tissue hypoxia, leukocyte activation, oxidative stress, and inflammatory mediators induced by renal I/R have been implicated in ARF-associated cardiac dysfunction.2 Curcumin, the principal curcuminoid of turmeric, was found to possess anti-inflammatory and antioxidative capacity and was shown effective in protecting I/R-induced remote organ injury.3 However, the protective efficacy of curcumin pretreatment against renal I/R-induced myocardial injury and impairment of cardiac function were not studied. In this study, we
assessed the cardiac function using simultaneously recorded left ventricular (LV) pressure-volume (PV) loops and parameters including LV stroke volume (SV), ejection fraction
From the Department of Medicine (Y.-C.Y., J.-C.W., J.-J.W.), School of Medicine, Fu Jen Catholic University, New Taipei City, and the Division of Cardiovascular Surgery (T.-H.C.), Cathay General Hospital, Taipei, Taiwan. Supported by an operating grant (100CGH-FJU-15) from the Cathy General Hospital (Taipei) and partially supported by NSC 100-2320-B-030-005 from the National Science Council Taiwan to Dr. Jiun-Jr Wang. Address reprint requests to Jiun-Jr Wang, PhD, Assistant Professor, School of Medicine, Fu Jen Catholic University, No. 510, Zhongzheng Rd., Xinzhuang Dist., New Taipei City, 24205, Taiwan. E-mail: [email protected]
0041-1345/13/$esee front matter http://dx.doi.org/10.1016/j.transproceed.2013.09.006
ª 2013 by Elsevier Inc. All rights reserved. 360 Park Avenue South, New York, NY 10010-1710
3546
Transplantation Proceedings, 45, 3546e3549 (2013)
CURCUMIN TREATMENT
(EF), systolic pressure (SP), and end-systolic-pressurevolume-relation (ESPVR). Renal I/R-induced myocardial injury was assessed based on serum creatine kinase muscle brain fraction (CK-MB) and Troponin I (cTnI) level, and kidney injury was assessed based on serum blood urea nitrogen (BUN) and creatinine level. The status of myocardial inflammation and lipid peroxidation were examined by tissue levels of tumor necrosis factor (TNF)-a and malondialdehyde (MDA), respectively. MATERIALS AND METHODS Sprague-Dawley rats (250e300 g) were anesthetized with pentobarbital sodium (35 mg/kg, intraperitoneally [i.p.]), followed by a tracheotomy, where a sterile polystyrene cannula (PE240) was inserted in the trachea and connected to a small animal mechanical ventilator (Topo, Kent Scientific, Torrington, CT, United States). The simultaneous LV pressure and volume were measured using a high-fidelity conductance-based pressure-volume transducer (FTS1212B-4518, Scisense Technology, London, ON, Canada), introduced through the right carotid artery and advanced into the LV chamber. Real-time data were collected using a data acquisition system (MP150, Biopac Systems Inc, Goleta, Calif, United States) and recorded at 2000 Hz. A femoral artery was cannulated with a polyethylene catheter (PE50), through which a blood sample was drawn for laboratory evaluations. The temperature was maintained at 37 C by a heating blanket. At the end of study, animals were humanely killed by an overdose of anesthetic (100 mg/kg, i.p.).
Induction of I/R A small midline incision was performed at the abdomen, through which bilateral occlusion of renal pedicles was performed to induce ischemia for 45 minutes, followed by 3 hours of reperfusion by releasing the occluders.
Assessment of Cardiac Function The cardiac function was evaluated based on LV SV, EF, SP, and ESPVR at different phases during the experiment, at baseline, 40 minutes of ischemia, and 1 and 3 hours of reperfusion. The LV contractility was evaluated by the slope of the ESPVR, composed of a sequence of reducing PV loops, generated by decreasing the LV preload through temporarily compressing the subdiaphragmatic inferior vena cava through an abdominal incision.
Assessments of Serum Levels of CK-MB, cTnI, Creatinine, and BUN Blood samples, collected at baseline, 45 minutes of ischemia, and 3 hours of reperfusion, were centrifuged at 2000 rpm for 10 minutes. Serum cTnI level was measured in the supernatant by a commercial kit and an i-STAT analyzer (Abbot Point of Care Inc., Princeton, NJ, United States), and levels of CK-MB, creatinine, and BUN were examined in the top supernatant using commercial kits via a Fuji DRI-CHEM 4000i system (Fujifilm Co., Tokyo, Japan).
Assessments of Myocardial TNF-a and MDA At the end of study, a section of LV anterior wall was dissected, lysed, and homogenized. The levels of TNF-a and MDA were analyzed in the supernatants of tissue homogenates using enzymelinked immunosorbent assay (ELISA) kits (ab100785 and
3547 Table 1. The Cardiac Function Evaluated Based on SV, EF, SP, and the ESPVR at Different Times During the Experiment Group
Control Baseline 40 min 1h 3h I/R Baseline 40 min I 1hR 3hR Curcumin þI/R Baseline 40 min I 1hR 3hR
SV (mL)
EF (%)
SP (mm Hg)
ESPVR (mm Hg/mL)
278 268 274 261
28 31 34 22
70 65 65 68
3 4 3 5
108 102 96 98
10 4 10 8
0.86 0.82 0.85 0.84
0.06 0.06 0.05 0.09
264 247 242 207
33 31 28 41*
69 60 54 38
2 4 6* 10*
112 88 74 68
5 11 9* 12*
0.86 0.83 0.66 0.34
0.07 0.07 0.09 0.14*
266 246 254 229
25 29 32 43*
67 62 60 56
3 6 6 5*†
109 92 90 76
7 9 6 7*†
0.91 0 84 0.71 0.55
0.08 0 06 0.10 0.08*†
Abbreviations: I, ischemia; R, reperfusion. *P < .05 vs sham group. † P < .05 vs renal I/R group.
ab118970, Abcam, Cambridge, Mass, United States) according to manufacturer’s guidelines.
Experimental Design Thirty rats were evenly divided into a renal I/R group, a sham group (same surgery except vessel occlusions), and a group with 2 days of oral pretreatment with curcumin (12.5 mg/kg/d) prior to renal I/R injury.
Data Analysis Data were presented as mean SEM. Comparisons across more than 2 groups were analyzed with one-way analysis of variance, followed by Scheffe’s method. Paired Student t test was used to compare the baseline values with those of the post-I/R within each group. P < .05 was considered statistically significant.
RESULTS
During renal ischemia, SV, SP, and EF reduced slightly, and ESPVR was relatively unchanged as compared with the baseline. During early reperfusion, SV, EF, SP, and ESPVR all increased temporarily and then decreased steadily until the end of the study (Table 1). Markedly elevated serum CK-MB (Fig 1A) and cTnI (Fig 1B) levels were found at the end of reperfusion, but not during the ischemic phase as compared with the baseline, suggested myocardial injury mainly occurred during reperfusion. Renal I/R injury increased MDA (Fig 1C) and TNF-alpha (Fig 1D) levels in the heart tissue as compared with the sham group, indicating elevated myocardial inflammation and oxidative stress. In contrast, 2 days of curcumin pretreatment effectively attenuated deterioration of cardiac function and reduced heart and kidney injury (Fig 1E and 1F), as well as myocardial inflammation and oxidative stress. DISCUSSION
In this study, we observed that renal ischemia through bilateral occlusions of renal pedicles, by itself, had limited
3548
CHEN, YANG, WANG ET AL
A
B
C
D
E
F
Fig 1. Renal I/R injury significantly increased serum CK-MB (A), cTnI (B), heart tissue MDA (C), and TNF-a (D), along with BUN (E) and creatinine (F). Curcumin pretreatment effective reduced myocardial and kidney injury. (*P < .05).
impact on cardiac function and myocardial injury; more serious injuries were observed following 3 hours of reperfusion. In addition, we observed marked elevation in oxidative stress and inflammation in the post-I/R myocardium. Hassoun et al1 postulated that remote organ injury induced by renal I/R injury is strongly associated with leukocyte activation and systemic inflammatory responses, manifested during reperfusion phase. Kelly4 found that protein contents of TNF-a and interleukin-1 and the expression of intercellular adhesion molecule-1 messenger RNA in the heart were markedly increased subsequent to renal I/R injury. Furthermore, Chiang et al5 demonstrated that, in addition to inflammatory cytokines, leukocyte activation following renal I/R injury increased the degree of tissue oxidative stress, leading to kidney tissue injury and apoptosis. We demonstrated that 2 days of curcumim pretreatment effectively attenuated myocardial inflammation and oxidative stress. The improvements in post-renal I/R cardiac function and myocardial injury were most
likely associated with the anti-inflammatory and antioxidative capacity of curcumin. Aggarwal and Harikumar2 showed that curcumin is an effective antioxidant that protects endothelial cells through heme oxygenase-1 synthesis.6 Kim et al7 demonstrated that curcumin is a potent anti-inflammatory agent, protecting tissues against cell damage through the modulation of p38 and signal transducer and activator of transcription 3 (STAT-3) and TNF-a in human endothelial cells. In addition, Izem-Meziane et al8 showed that curcumin can ameliorate mitochondria damage and cardiac dysfunction as a result of reperfusion injury, through regulating mitochondrial permeability transition.8 We concluded that 45 minutes of renal ischemia did not cause significant damage to myocardium or cardiac function, impairments of cardiac function, and myocardial injury after 3 hours of reperfusion was strongly associated with renal I/R-induced myocardial inflammation and increasing oxidative stress. Curcumin pretreatment effectively attenuated post-I/R myocardial injury and
CURCUMIN TREATMENT
improved cardiac function through reducing myocardial inflammation and lipid peroxidation in the heart tissue. REFERENCES 1. Hassoun HT, Grigoryev DN, Lie ML, et al. Ischemic acute kidney injury induces a distant organ functional and genomic response distinguishable from bilateral nephrectomy. Am J Physiol Renal Physiol. 2007;293(1):F30eF40. 2. Aggarwal BB, Harikumar KB. Potential therapeutic effects of curcumin, the anti-inflammatory agent, against neurodegenerative, cardiovascular, pulmonary, metabolic, autoimmune and neoplastic diseases. Int J Biochem Cell Biol. 2009;41(1):40e59. 3. Chen KH, Chao D, Liu CF, et al. Curcumin attenuates airway hyperreactivity induced by ischemia-reperfusion of the pancreas in rats. Transplant Proc. 2010;42(3):744e747.
3549 4. Kelly KJ. Distant effects of experimental renal ischemia/ reperfusion injury. J Am Soc Nephrol. 2003;14(6):1549e1558. 5. Chiang WC, Chien CT, Lin WW, et al. Early activation of bradykinin B2 receptor aggravates reactive oxygen species generation and renal damage in ischemia/reperfusion injury. Free Radic Biol Med. 2006;41(8):1304e1314. 6. Motterlini R, Foresti R, Bassi R, et al. Curcumin, an antioxidant and anti-inflammatory agent, induces heme oxygenase-1 and protects endothelial cells against oxidative stress. Free Radic Biol Med. 2000;28(8):1303e1312. 7. Kim YS, Ahn Y, Hong MH, et al. Curcumin attenuates inflammatory responses of TNF-alpha-stimulated human endothelial cells. J Cardiovasc Pharmacol. 2007;50(1):41e49. 8. Izem-Meziane M, Djerdjouri B, Rimbaud S, et al. Catecholamine-induced cardiac mitochondrial dysfunction and mPTP opening: protective effect of curcumin. Am J Physiol Heart Circ Physiol. 2012;302(3):H665eH674.
R
ENAL ischemia and reperfusion (I/R) injury frequently occurred following sepsis, vasoconstrictive drugs, and kidney transplantation, which is the major cause of acute renal failure (ARF) resulting in significant morbidity and mortality.1 However, cardiac failure, instead of renal failure itself, has been observed as the primary cause of death in this clinical condition. Myocardial depression factors, tissue hypoxia, leukocyte activation, oxidative stress, and inflammatory mediators induced by renal I/R have been implicated in ARF-associated cardiac dysfunction.2 Curcumin, the principal curcuminoid of turmeric, was found to possess anti-inflammatory and antioxidative capacity and was shown effective in protecting I/R-induced remote organ injury.3 However, the protective efficacy of curcumin pretreatment against renal I/R-induced myocardial injury and impairment of cardiac function were not studied. In this study, we
assessed the cardiac function using simultaneously recorded left ventricular (LV) pressure-volume (PV) loops and parameters including LV stroke volume (SV), ejection fraction
From the Department of Medicine (Y.-C.Y., J.-C.W., J.-J.W.), School of Medicine, Fu Jen Catholic University, New Taipei City, and the Division of Cardiovascular Surgery (T.-H.C.), Cathay General Hospital, Taipei, Taiwan. Supported by an operating grant (100CGH-FJU-15) from the Cathy General Hospital (Taipei) and partially supported by NSC 100-2320-B-030-005 from the National Science Council Taiwan to Dr. Jiun-Jr Wang. Address reprint requests to Jiun-Jr Wang, PhD, Assistant Professor, School of Medicine, Fu Jen Catholic University, No. 510, Zhongzheng Rd., Xinzhuang Dist., New Taipei City, 24205, Taiwan. E-mail: [email protected]
0041-1345/13/$esee front matter http://dx.doi.org/10.1016/j.transproceed.2013.09.006
ª 2013 by Elsevier Inc. All rights reserved. 360 Park Avenue South, New York, NY 10010-1710
3546
Transplantation Proceedings, 45, 3546e3549 (2013)
CURCUMIN TREATMENT
(EF), systolic pressure (SP), and end-systolic-pressurevolume-relation (ESPVR). Renal I/R-induced myocardial injury was assessed based on serum creatine kinase muscle brain fraction (CK-MB) and Troponin I (cTnI) level, and kidney injury was assessed based on serum blood urea nitrogen (BUN) and creatinine level. The status of myocardial inflammation and lipid peroxidation were examined by tissue levels of tumor necrosis factor (TNF)-a and malondialdehyde (MDA), respectively. MATERIALS AND METHODS Sprague-Dawley rats (250e300 g) were anesthetized with pentobarbital sodium (35 mg/kg, intraperitoneally [i.p.]), followed by a tracheotomy, where a sterile polystyrene cannula (PE240) was inserted in the trachea and connected to a small animal mechanical ventilator (Topo, Kent Scientific, Torrington, CT, United States). The simultaneous LV pressure and volume were measured using a high-fidelity conductance-based pressure-volume transducer (FTS1212B-4518, Scisense Technology, London, ON, Canada), introduced through the right carotid artery and advanced into the LV chamber. Real-time data were collected using a data acquisition system (MP150, Biopac Systems Inc, Goleta, Calif, United States) and recorded at 2000 Hz. A femoral artery was cannulated with a polyethylene catheter (PE50), through which a blood sample was drawn for laboratory evaluations. The temperature was maintained at 37 C by a heating blanket. At the end of study, animals were humanely killed by an overdose of anesthetic (100 mg/kg, i.p.).
Induction of I/R A small midline incision was performed at the abdomen, through which bilateral occlusion of renal pedicles was performed to induce ischemia for 45 minutes, followed by 3 hours of reperfusion by releasing the occluders.
Assessment of Cardiac Function The cardiac function was evaluated based on LV SV, EF, SP, and ESPVR at different phases during the experiment, at baseline, 40 minutes of ischemia, and 1 and 3 hours of reperfusion. The LV contractility was evaluated by the slope of the ESPVR, composed of a sequence of reducing PV loops, generated by decreasing the LV preload through temporarily compressing the subdiaphragmatic inferior vena cava through an abdominal incision.
Assessments of Serum Levels of CK-MB, cTnI, Creatinine, and BUN Blood samples, collected at baseline, 45 minutes of ischemia, and 3 hours of reperfusion, were centrifuged at 2000 rpm for 10 minutes. Serum cTnI level was measured in the supernatant by a commercial kit and an i-STAT analyzer (Abbot Point of Care Inc., Princeton, NJ, United States), and levels of CK-MB, creatinine, and BUN were examined in the top supernatant using commercial kits via a Fuji DRI-CHEM 4000i system (Fujifilm Co., Tokyo, Japan).
Assessments of Myocardial TNF-a and MDA At the end of study, a section of LV anterior wall was dissected, lysed, and homogenized. The levels of TNF-a and MDA were analyzed in the supernatants of tissue homogenates using enzymelinked immunosorbent assay (ELISA) kits (ab100785 and
3547 Table 1. The Cardiac Function Evaluated Based on SV, EF, SP, and the ESPVR at Different Times During the Experiment Group
Control Baseline 40 min 1h 3h I/R Baseline 40 min I 1hR 3hR Curcumin þI/R Baseline 40 min I 1hR 3hR
SV (mL)
EF (%)
SP (mm Hg)
ESPVR (mm Hg/mL)
278 268 274 261
28 31 34 22
70 65 65 68
3 4 3 5
108 102 96 98
10 4 10 8
0.86 0.82 0.85 0.84
0.06 0.06 0.05 0.09
264 247 242 207
33 31 28 41*
69 60 54 38
2 4 6* 10*
112 88 74 68
5 11 9* 12*
0.86 0.83 0.66 0.34
0.07 0.07 0.09 0.14*
266 246 254 229
25 29 32 43*
67 62 60 56
3 6 6 5*†
109 92 90 76
7 9 6 7*†
0.91 0 84 0.71 0.55
0.08 0 06 0.10 0.08*†
Abbreviations: I, ischemia; R, reperfusion. *P < .05 vs sham group. † P < .05 vs renal I/R group.
ab118970, Abcam, Cambridge, Mass, United States) according to manufacturer’s guidelines.
Experimental Design Thirty rats were evenly divided into a renal I/R group, a sham group (same surgery except vessel occlusions), and a group with 2 days of oral pretreatment with curcumin (12.5 mg/kg/d) prior to renal I/R injury.
Data Analysis Data were presented as mean SEM. Comparisons across more than 2 groups were analyzed with one-way analysis of variance, followed by Scheffe’s method. Paired Student t test was used to compare the baseline values with those of the post-I/R within each group. P < .05 was considered statistically significant.
RESULTS
During renal ischemia, SV, SP, and EF reduced slightly, and ESPVR was relatively unchanged as compared with the baseline. During early reperfusion, SV, EF, SP, and ESPVR all increased temporarily and then decreased steadily until the end of the study (Table 1). Markedly elevated serum CK-MB (Fig 1A) and cTnI (Fig 1B) levels were found at the end of reperfusion, but not during the ischemic phase as compared with the baseline, suggested myocardial injury mainly occurred during reperfusion. Renal I/R injury increased MDA (Fig 1C) and TNF-alpha (Fig 1D) levels in the heart tissue as compared with the sham group, indicating elevated myocardial inflammation and oxidative stress. In contrast, 2 days of curcumin pretreatment effectively attenuated deterioration of cardiac function and reduced heart and kidney injury (Fig 1E and 1F), as well as myocardial inflammation and oxidative stress. DISCUSSION
In this study, we observed that renal ischemia through bilateral occlusions of renal pedicles, by itself, had limited
3548
CHEN, YANG, WANG ET AL
A
B
C
D
E
F
Fig 1. Renal I/R injury significantly increased serum CK-MB (A), cTnI (B), heart tissue MDA (C), and TNF-a (D), along with BUN (E) and creatinine (F). Curcumin pretreatment effective reduced myocardial and kidney injury. (*P < .05).
impact on cardiac function and myocardial injury; more serious injuries were observed following 3 hours of reperfusion. In addition, we observed marked elevation in oxidative stress and inflammation in the post-I/R myocardium. Hassoun et al1 postulated that remote organ injury induced by renal I/R injury is strongly associated with leukocyte activation and systemic inflammatory responses, manifested during reperfusion phase. Kelly4 found that protein contents of TNF-a and interleukin-1 and the expression of intercellular adhesion molecule-1 messenger RNA in the heart were markedly increased subsequent to renal I/R injury. Furthermore, Chiang et al5 demonstrated that, in addition to inflammatory cytokines, leukocyte activation following renal I/R injury increased the degree of tissue oxidative stress, leading to kidney tissue injury and apoptosis. We demonstrated that 2 days of curcumim pretreatment effectively attenuated myocardial inflammation and oxidative stress. The improvements in post-renal I/R cardiac function and myocardial injury were most
likely associated with the anti-inflammatory and antioxidative capacity of curcumin. Aggarwal and Harikumar2 showed that curcumin is an effective antioxidant that protects endothelial cells through heme oxygenase-1 synthesis.6 Kim et al7 demonstrated that curcumin is a potent anti-inflammatory agent, protecting tissues against cell damage through the modulation of p38 and signal transducer and activator of transcription 3 (STAT-3) and TNF-a in human endothelial cells. In addition, Izem-Meziane et al8 showed that curcumin can ameliorate mitochondria damage and cardiac dysfunction as a result of reperfusion injury, through regulating mitochondrial permeability transition.8 We concluded that 45 minutes of renal ischemia did not cause significant damage to myocardium or cardiac function, impairments of cardiac function, and myocardial injury after 3 hours of reperfusion was strongly associated with renal I/R-induced myocardial inflammation and increasing oxidative stress. Curcumin pretreatment effectively attenuated post-I/R myocardial injury and
CURCUMIN TREATMENT
improved cardiac function through reducing myocardial inflammation and lipid peroxidation in the heart tissue. REFERENCES 1. Hassoun HT, Grigoryev DN, Lie ML, et al. Ischemic acute kidney injury induces a distant organ functional and genomic response distinguishable from bilateral nephrectomy. Am J Physiol Renal Physiol. 2007;293(1):F30eF40. 2. Aggarwal BB, Harikumar KB. Potential therapeutic effects of curcumin, the anti-inflammatory agent, against neurodegenerative, cardiovascular, pulmonary, metabolic, autoimmune and neoplastic diseases. Int J Biochem Cell Biol. 2009;41(1):40e59. 3. Chen KH, Chao D, Liu CF, et al. Curcumin attenuates airway hyperreactivity induced by ischemia-reperfusion of the pancreas in rats. Transplant Proc. 2010;42(3):744e747.
3549 4. Kelly KJ. Distant effects of experimental renal ischemia/ reperfusion injury. J Am Soc Nephrol. 2003;14(6):1549e1558. 5. Chiang WC, Chien CT, Lin WW, et al. Early activation of bradykinin B2 receptor aggravates reactive oxygen species generation and renal damage in ischemia/reperfusion injury. Free Radic Biol Med. 2006;41(8):1304e1314. 6. Motterlini R, Foresti R, Bassi R, et al. Curcumin, an antioxidant and anti-inflammatory agent, induces heme oxygenase-1 and protects endothelial cells against oxidative stress. Free Radic Biol Med. 2000;28(8):1303e1312. 7. Kim YS, Ahn Y, Hong MH, et al. Curcumin attenuates inflammatory responses of TNF-alpha-stimulated human endothelial cells. J Cardiovasc Pharmacol. 2007;50(1):41e49. 8. Izem-Meziane M, Djerdjouri B, Rimbaud S, et al. Catecholamine-induced cardiac mitochondrial dysfunction and mPTP opening: protective effect of curcumin. Am J Physiol Heart Circ Physiol. 2012;302(3):H665eH674.
