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Inhibitory Effects of Omega-3 PUFAs on Cardiac Fibrosis In Vivo and In Vitro
S22 Journal of Cardiac Failure Vol. 15 No. 6S Suppl. 2009 Results: 11 patients (4 female) were identified from Aug 2008 to March 2009, with mean age 58 6 14 years, LVEF 23 6 17%, Cr 1.75 6 1 mg/dL, and BUN 46 6 25 mg/dL. 7 (63.6%) had no ascites, 4 had small to moderate amounts of ascites. Of the 7 patients without ascites, 5 had elevated IAP. 3 out of 4 patients with ascites had elevated IAP. Thus 62.5% of patients with elevated IAP had no ascites. 2 patients had concurrent intra-abdominal hematoma which likely contributed to the observed IAP. Of these, 1 did not have ascites and the other only had a small amount of ascites. Both had moderate to severe renal dysfunction (Cr 2.51 to 3.79).
Placebo (n549) Delta BW [g] Delta fat [g] Delta lean [g] EF [%] FS [%] SV [mL] nmol ROS/mg protein/min Proteasome activity [mU]
-50.2 -11.2 -37.1 57.3 31.5 105.6 169.51
6 6 6 6 6 6 6
2.2 0.4 2.0 2.7 1.9 8.4 52.5
254.8 6 73.2
Oxy 4mg/kg/d (n511) -15.4 -6.8 -10.6 71.3 42.5 152.3 25.6
6 6 6 6 6 6 6
14.2*** 2.6** 10.4*** 4.1* 3.6* 24.3* 6.5**
37.8 6 12.7*
Oxy 40mg/kg/d (n512) -45.3 -14.4 -32.4 68.8 41.3 129.5 17.3
6 6 6 6 6 6 6
11.8 1.9* 9.6 5.3 5.2* 34.9 8.3*
112.2 6 69.3
Allo 4mg/kg/d (n512) -31.9 -9.9 -23.7 61.3 38.6 150.1 38.2
6 6 6 6 6 6 6
14.9 2 12.2 6.5 4.9 34.4 5.7*
84.3 6 33.4
Allo 40mg/kg/d (n511) -26.4 -8.1 -19.9 75.9 48.1 120.1 31.2
6 6 6 6 6 6 6
17.1* 2.9 12.6* 5.6** 5.3** 25 12.6*
97.1 6 42.3
BW: body weight, * p!0.05, ** p5!0.01, ***p!0.001 improved vs placebo.
Conclusion: In this retrospective review, ascites was not present in most of the HF patients with elevated IAP. Elevated IAP is important as it may impair renal function. Although ascites is a recognized cause of elevated IAP, the mere presence of elevated IAP does not equate to the presence of ascites in HF patients. Visceral edema may be the reason for this phenomenon and further investigation should be performed to study this.
cancer cachexia. Rats (weight approx. 200 g) were inoculated with AH-130 hepatoma cells and treated with allopurinol (Allo, 4 and 40mg/kg/d), oxypurinol (Oxy, 4 and 40mg/kg/d), or placebo. Cardiac function was assessed before inoculation and on day 11 of the 16-day protocol. Weight and body composition (NMR-scan) were assessed on day 0 and day 16 after sacrifice. XO-generated reactive oxygen species (ROS) were assessed by EPR after purification of XO. Proteasome activity was measured by turnover of the flurogenic substrate Suc-LLVY-AMC. Weight loss consisting of wasting of both fat and lean tissue was significantly reduced by 4 mg/kg/d Oxy and by the 10 times higher dose of Allo, but not by low dose (LD) Allo and high dose (HD) oxy. Cardiac function was significantly improved by XO inhibition. Treatment with Allo and Oxy not only reduced XO generated ROS, but also reduced the proteasome activity. As a result, survival was significantly improved by LD Oxy (HR: 0.37, 95%CI: 0.17-0.79, p50.011) and HD Allo (HR: 0.40, 95%CI: 0.19-0.86, p50.019). Conclusion: Inhibition of XO can reduce tissue wasting as well as improve cardiac function and survival in a model of cancer cachexia. A low dose of the second generation XO-inhibitor oxypurinol had superior properties to allopurinol.
061 059 Impact of Serum Iron Availability on Hemoglobin and Hemdoynamics in Patients with Advanced Heart Failure Micha T. Maeder1,2, David M. Kaye1,2; 1Heart Failure Research Group, Baker IDI Heart and Diabetes Institute, Melbourne, Victoria, Australia; 2Heart Center, Alfred Hospital, Melbourne, Victoria, Australia Introduction: Anemia is a marker of poor outcome in heart failure (HF) patients. However, the causes of anemia in HF and the mechanisms underlying its association with poor outcome are unclear. Hypothesis: We hypothesized that measures of serum iron availability are related to hemoglobin (Hb) levels and that measures of serum iron availability and Hb respectively are related to hemodynamics in patients with advanced HF. Methods: Hemoglobin, hematinic levels, estimated glomerular filtration rate (eGFR), and high-sensitivity C-reactive protein (hs-CRP) were assessed in 16 patients with advanced HF [median (interquartile range) left ventricular ejection fraction 20 (16-25)%] undergoing right heart catheterization. Results: Serum iron (r50.68; p50.004) and transferrin saturation (TFS; r50.69; p50.003) were directly related to Hb. In contrast, there was no significant correlation between Hb and ferritin, folate, vitamin B12, eGFR, and hs-CRP respectively. Hemoglobin was directly related to mean arterial pressure (MAP; r50.72; p50.002) and stroke volume (SV; r50.55; p50.03) and inversely related to heart rate (HR; r5-0.58; p50.02). Similar or even stronger correlations were observed between iron and TFS respectively and MAP (r50.69; p50.003 / r50.75; p50.001), SV (r50.72; p50.002 / r50.71; p50.002), and HR (r5-0.73; p50.001 / r5-0.66; p50.005). Conclusions: In patients with advanced HF, Hb levels are strongly related to measures of serum iron availability suggesting that absolute (low iron stores) or relative (iron sequestration) iron deficiency contributes to anemia in HF. Serum iron availability is more closely related to SV than Hb is, suggesting that iron availability might affect cardiac performance by Hb-related and non-Hb-related effects.
060 The Xanthine Oxidase Inhibitors Oxypurinol and Allopurinol Reduce Wasting and Improve Cardiac Function in Experimental Cancer Cachexia Jochen Springer1, Anika Hartmann1, Sandra Palus1, Volker Adams2, Ruediger von Harsdorf3, Stefan D. Anker1, Wolfram Doehner1; 1Applied Cachexia Research, CCR, Charite Medical School, Germany; 2Heart Center Leipzig, University Leipzig, Leipzig, Germany; 3Division of Cardiology, University Network Hospitals, Toronto, ON, Canada Cachexia is a co-morbidity in cancer patients. In chronic heart failure, high serum uric acid levels are associated with metabolic illness and poor survival. We hypothesised that xanthine oxidase (XO) inhibition reduces tissue wasting and improve survival in
Inhibitory Effects of Omega-3 PUFAs on Cardiac Fibrosis In Vivo and In Vitro Jinghai Chen, Gregory Shearer, Timothy D. O’Connell, A. Martin Gerdes, William S. Harris, Dajun Wang; Cardiovascular Health Research Center, Sanford Research/ USD, Sioux Falls, SD Introduction: Many studies have shown that omega-3 polyunsaturated fatty acids (u-3 PUFAs) have beneficial effects on cardiovascular disease but little is known about the effects of u-3 PUFAs on cardiac fibrosis, which is the major cause of cardiac remodeling leading to hear failure. Hypothesis: u-3 PUFAs will inhibit cardiac fibrosis and improve heart function. Methods: Ten weeks old male C57/ B6 mice were fed control diet (corn oil, u-6 PUFAs; n520) or Fish oil diet (FO) 1% energy from EPAþDHA (n520). After 8 weeks on the assigned diet, Sham or Transverse Aortic Constriction (TAC) surgery will be randomly performed on each diet group. The groups allocated in a 2x2 factorial design into 4 groups as (1) Cont/Sham, (2) Cont/TAC, (3) FO/Sham, (4) FO/TAC. After 4 weeks of continued dietary, myocardial fibrosis was determined by picro-sirius red staining for collagen in the middle of left ventricle. The u-3 index (the sum of red blood cell EPA þ DHA) were determined by gas chromatography. Echocardiography (Echo) were also performed on mice one day before and 2- and 4-weeks post-surgery. In vitro experiments were performed in adult cardiac fibroblasts (CFs) isolated from the heart of male C57BL/6 mice. CFs collagen synthesis, differentiation to myofibroblasts and proliferation were determined by 3H-proline incorporation, a-smooth muscle actin (aSMA) staining and MTS assay, respectively. Results: Compare to Cont/sham group, in Cont/TAC group, Fractional Shortening (FS) were decreased significantly (p!0.01) as well as myocardial fibrosis was significantly increased in (p!0.01) at 4 weeks. However, FO treatment rescued the effects of TAC stress as FS remarkably reverse (FO/ TAC 45%610% vs Cont/TAC 28%610%, p!0.01) and myocardial fibrosis significantly decreased (FO/ TAC 8.9%62.8% vs Cont/TAC 14.1%64.1%, p!0.01). In addition, the u-3 index was notably increased in FO diet group (FO 15.17%60.4% vs Cont 3.9%6 0.6%, p!0.01). In vitro, EPA and DHA (10 mM) significantly blocked the pro-fibrotic effects of TGF- b1 (1ng/mL) on collagen synthesis while arachidonic acid (AA) and oleic acid (OA) did not. Both EPA and DHA suppressed TGF- b1-induced differentiation of CFs to myofibroblasts. Furthermore, EPA and DHA (1 to 50 mM) inhibited CFs proliferation induce by both Angiotensin II (100 nM) and TGF- b1 (1ng/mL) more than AA (p!0.01) and OA (p!0.05) did. Conclusion: Both in vivo and in vitro. u-3 PUFAs have the potential to inhibit the development of cardiac fibrosis.
062 Characterization of a Non-Surgical Mouse Model of Acute Non-Ischemic Cardiomyopathy Keith A. Youker1, Rene Celis1, Carlos Orrego1, Jerry D. Estep1, Dale Hamilton2, Guillermo Torre-Amione1; 1Cardiology, The Methodist DeBakey Heart & Vascular
Placebo (n549) Delta BW [g] Delta fat [g] Delta lean [g] EF [%] FS [%] SV [mL] nmol ROS/mg protein/min Proteasome activity [mU]
-50.2 -11.2 -37.1 57.3 31.5 105.6 169.51
6 6 6 6 6 6 6
2.2 0.4 2.0 2.7 1.9 8.4 52.5
254.8 6 73.2
Oxy 4mg/kg/d (n511) -15.4 -6.8 -10.6 71.3 42.5 152.3 25.6
6 6 6 6 6 6 6
14.2*** 2.6** 10.4*** 4.1* 3.6* 24.3* 6.5**
37.8 6 12.7*
Oxy 40mg/kg/d (n512) -45.3 -14.4 -32.4 68.8 41.3 129.5 17.3
6 6 6 6 6 6 6
11.8 1.9* 9.6 5.3 5.2* 34.9 8.3*
112.2 6 69.3
Allo 4mg/kg/d (n512) -31.9 -9.9 -23.7 61.3 38.6 150.1 38.2
6 6 6 6 6 6 6
14.9 2 12.2 6.5 4.9 34.4 5.7*
84.3 6 33.4
Allo 40mg/kg/d (n511) -26.4 -8.1 -19.9 75.9 48.1 120.1 31.2
6 6 6 6 6 6 6
17.1* 2.9 12.6* 5.6** 5.3** 25 12.6*
97.1 6 42.3
BW: body weight, * p!0.05, ** p5!0.01, ***p!0.001 improved vs placebo.
Conclusion: In this retrospective review, ascites was not present in most of the HF patients with elevated IAP. Elevated IAP is important as it may impair renal function. Although ascites is a recognized cause of elevated IAP, the mere presence of elevated IAP does not equate to the presence of ascites in HF patients. Visceral edema may be the reason for this phenomenon and further investigation should be performed to study this.
cancer cachexia. Rats (weight approx. 200 g) were inoculated with AH-130 hepatoma cells and treated with allopurinol (Allo, 4 and 40mg/kg/d), oxypurinol (Oxy, 4 and 40mg/kg/d), or placebo. Cardiac function was assessed before inoculation and on day 11 of the 16-day protocol. Weight and body composition (NMR-scan) were assessed on day 0 and day 16 after sacrifice. XO-generated reactive oxygen species (ROS) were assessed by EPR after purification of XO. Proteasome activity was measured by turnover of the flurogenic substrate Suc-LLVY-AMC. Weight loss consisting of wasting of both fat and lean tissue was significantly reduced by 4 mg/kg/d Oxy and by the 10 times higher dose of Allo, but not by low dose (LD) Allo and high dose (HD) oxy. Cardiac function was significantly improved by XO inhibition. Treatment with Allo and Oxy not only reduced XO generated ROS, but also reduced the proteasome activity. As a result, survival was significantly improved by LD Oxy (HR: 0.37, 95%CI: 0.17-0.79, p50.011) and HD Allo (HR: 0.40, 95%CI: 0.19-0.86, p50.019). Conclusion: Inhibition of XO can reduce tissue wasting as well as improve cardiac function and survival in a model of cancer cachexia. A low dose of the second generation XO-inhibitor oxypurinol had superior properties to allopurinol.
061 059 Impact of Serum Iron Availability on Hemoglobin and Hemdoynamics in Patients with Advanced Heart Failure Micha T. Maeder1,2, David M. Kaye1,2; 1Heart Failure Research Group, Baker IDI Heart and Diabetes Institute, Melbourne, Victoria, Australia; 2Heart Center, Alfred Hospital, Melbourne, Victoria, Australia Introduction: Anemia is a marker of poor outcome in heart failure (HF) patients. However, the causes of anemia in HF and the mechanisms underlying its association with poor outcome are unclear. Hypothesis: We hypothesized that measures of serum iron availability are related to hemoglobin (Hb) levels and that measures of serum iron availability and Hb respectively are related to hemodynamics in patients with advanced HF. Methods: Hemoglobin, hematinic levels, estimated glomerular filtration rate (eGFR), and high-sensitivity C-reactive protein (hs-CRP) were assessed in 16 patients with advanced HF [median (interquartile range) left ventricular ejection fraction 20 (16-25)%] undergoing right heart catheterization. Results: Serum iron (r50.68; p50.004) and transferrin saturation (TFS; r50.69; p50.003) were directly related to Hb. In contrast, there was no significant correlation between Hb and ferritin, folate, vitamin B12, eGFR, and hs-CRP respectively. Hemoglobin was directly related to mean arterial pressure (MAP; r50.72; p50.002) and stroke volume (SV; r50.55; p50.03) and inversely related to heart rate (HR; r5-0.58; p50.02). Similar or even stronger correlations were observed between iron and TFS respectively and MAP (r50.69; p50.003 / r50.75; p50.001), SV (r50.72; p50.002 / r50.71; p50.002), and HR (r5-0.73; p50.001 / r5-0.66; p50.005). Conclusions: In patients with advanced HF, Hb levels are strongly related to measures of serum iron availability suggesting that absolute (low iron stores) or relative (iron sequestration) iron deficiency contributes to anemia in HF. Serum iron availability is more closely related to SV than Hb is, suggesting that iron availability might affect cardiac performance by Hb-related and non-Hb-related effects.
060 The Xanthine Oxidase Inhibitors Oxypurinol and Allopurinol Reduce Wasting and Improve Cardiac Function in Experimental Cancer Cachexia Jochen Springer1, Anika Hartmann1, Sandra Palus1, Volker Adams2, Ruediger von Harsdorf3, Stefan D. Anker1, Wolfram Doehner1; 1Applied Cachexia Research, CCR, Charite Medical School, Germany; 2Heart Center Leipzig, University Leipzig, Leipzig, Germany; 3Division of Cardiology, University Network Hospitals, Toronto, ON, Canada Cachexia is a co-morbidity in cancer patients. In chronic heart failure, high serum uric acid levels are associated with metabolic illness and poor survival. We hypothesised that xanthine oxidase (XO) inhibition reduces tissue wasting and improve survival in
Inhibitory Effects of Omega-3 PUFAs on Cardiac Fibrosis In Vivo and In Vitro Jinghai Chen, Gregory Shearer, Timothy D. O’Connell, A. Martin Gerdes, William S. Harris, Dajun Wang; Cardiovascular Health Research Center, Sanford Research/ USD, Sioux Falls, SD Introduction: Many studies have shown that omega-3 polyunsaturated fatty acids (u-3 PUFAs) have beneficial effects on cardiovascular disease but little is known about the effects of u-3 PUFAs on cardiac fibrosis, which is the major cause of cardiac remodeling leading to hear failure. Hypothesis: u-3 PUFAs will inhibit cardiac fibrosis and improve heart function. Methods: Ten weeks old male C57/ B6 mice were fed control diet (corn oil, u-6 PUFAs; n520) or Fish oil diet (FO) 1% energy from EPAþDHA (n520). After 8 weeks on the assigned diet, Sham or Transverse Aortic Constriction (TAC) surgery will be randomly performed on each diet group. The groups allocated in a 2x2 factorial design into 4 groups as (1) Cont/Sham, (2) Cont/TAC, (3) FO/Sham, (4) FO/TAC. After 4 weeks of continued dietary, myocardial fibrosis was determined by picro-sirius red staining for collagen in the middle of left ventricle. The u-3 index (the sum of red blood cell EPA þ DHA) were determined by gas chromatography. Echocardiography (Echo) were also performed on mice one day before and 2- and 4-weeks post-surgery. In vitro experiments were performed in adult cardiac fibroblasts (CFs) isolated from the heart of male C57BL/6 mice. CFs collagen synthesis, differentiation to myofibroblasts and proliferation were determined by 3H-proline incorporation, a-smooth muscle actin (aSMA) staining and MTS assay, respectively. Results: Compare to Cont/sham group, in Cont/TAC group, Fractional Shortening (FS) were decreased significantly (p!0.01) as well as myocardial fibrosis was significantly increased in (p!0.01) at 4 weeks. However, FO treatment rescued the effects of TAC stress as FS remarkably reverse (FO/ TAC 45%610% vs Cont/TAC 28%610%, p!0.01) and myocardial fibrosis significantly decreased (FO/ TAC 8.9%62.8% vs Cont/TAC 14.1%64.1%, p!0.01). In addition, the u-3 index was notably increased in FO diet group (FO 15.17%60.4% vs Cont 3.9%6 0.6%, p!0.01). In vitro, EPA and DHA (10 mM) significantly blocked the pro-fibrotic effects of TGF- b1 (1ng/mL) on collagen synthesis while arachidonic acid (AA) and oleic acid (OA) did not. Both EPA and DHA suppressed TGF- b1-induced differentiation of CFs to myofibroblasts. Furthermore, EPA and DHA (1 to 50 mM) inhibited CFs proliferation induce by both Angiotensin II (100 nM) and TGF- b1 (1ng/mL) more than AA (p!0.01) and OA (p!0.05) did. Conclusion: Both in vivo and in vitro. u-3 PUFAs have the potential to inhibit the development of cardiac fibrosis.
062 Characterization of a Non-Surgical Mouse Model of Acute Non-Ischemic Cardiomyopathy Keith A. Youker1, Rene Celis1, Carlos Orrego1, Jerry D. Estep1, Dale Hamilton2, Guillermo Torre-Amione1; 1Cardiology, The Methodist DeBakey Heart & Vascular