N-Acetyl Cysteine improves the diabetic cardiac function: possible role of fibrosis inhibition

BMC Cardiovascular Disorders

Li-Jun Yuan¹  Yun-You Duan¹  Jing Ma¹  Chang-Yang Xing¹  Ming-Zhi Shen²  Xiao-Zhao Lu²  Cong Liu¹ 

[1] Department of Ultrasound Diagnostics, Tangdu Hospital, Fourth Military Medical University, #569 Xinsi Road, Xi’an 710038, Baqiao District, China;Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi’an, China

关键词: Cardiac function;    N-acetyl-L-cysteine;    Reactive oxygen species;    Fibrosis;    Diabetic cardiomyopathy;   

Others  :  1222699
DOI  :  10.1186/s12872-015-0076-3

 received in 2015-05-25, accepted in 2015-07-24,  发布年份 2015

【 摘 要 】

Background

Diabetic cardiomyopathy is one of the leading causes of death in diabetes mellitus (DM) patients. This study aimed to explore the therapeutic implication of N-acetyl-L-cysteine (NAC, an antioxidant and glutathione precursor) and the possible underlying mechanism.

Methods

Thirty five 12-week-old male C57BL/6 mice were included. Twenty-five diabetic mice were induced by intraperitoneal injection of streptozocin (STZ, 150 mg/kg, Sigma-Aldrich) dissolved in a mix of citrate buffer after overnight fast. Mice with a blood glucose level above 13.5 mmol/L were considered diabetic. As a non-DM (diabetic) control, mice were injected with equal volume of citrate buffer. The 25 diabetic mice were divided into 5 groups with 5 animals in each group: including DM (diabetes without NAC treatment), and 4 different NAC treatment groups, namely NAC1, NAC3, NAC5 and NAC7, with the number defining the start time point of NAC treatment. In the 10 non-DM mice, mice were either untreated (Ctrl) or treated with NAC for 5 weeks (NAC only). Echocardiography was performed 12 weeks after STZ injection. Heart tissue were collected after echocardiography for Hematoxylin Eosin (HE) and Trichrome staining and ROS staining. Cardiac fibroblast cells were isolated, cultured and treated with high glucose plus NAC or the vehicle. qPCR analysis and CCK-8 assay were performed to observe fibrotic gene expression and cell proliferation.

Results

We found that both cardiac systolic function and diastolic function were impaired, coupled with excessive reactive oxygen stress and cardiac fibrosis 12 weeks after STZ induction. NAC significantly reduced ROS generation and fibrosis, together with improved cardiac systolic function and diastolic function. Strikingly, NAC1 treatment, which had the earlier and longer treatment, produced significant improvement of cardiac function and less fibrosis. In the cardiac fibroblasts, NAC blocked cardiac fibroblast proliferation and collagen synthesis induced by hyperglycemia.

Conclusions

Our study indicates that NAC treatment in diabetes effectively protects from diabetic cardiomyopathy, possibly through inhibiting the ROS production and fibrosis, which warrants further clarification.

【 授权许可】

   
2015 Liu et al.

【 图 表 】

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【 参考文献 】

• [1]Whiting DR, Guariguata L, Weil C, Shaw J: IDF diabetes atlas: global estimates of the prevalence of diabetes for 2011 and 2030. Diabetes Res Clin Pract 2011, 94(3):311-321.
• [2]Rajesh M, Mukhopadhyay P, Batkai S, Mukhopadhyay B, Patel V, Hasko G, et al.: Xanthine oxidase inhibitor allopurinol attenuates the development of diabetic cardiomyopathy. J Cell Mol Med 2009, 13(8B):2330-2341.
• [3]Takeda N, Manabe I: Cellular Interplay between Cardiomyocytes and Nonmyocytes in Cardiac Remodeling. Int J Inflamm 2011, 2011:535241.
• [4]Brown RD, Ambler SK, Mitchell MD, Long CS: The cardiac fibroblast: therapeutic target in myocardial remodeling and failure. Annu Rev Pharmacol Toxicol 2005, 45:657-687.
• [5]Asbun J, Villarreal FJ: The pathogenesis of myocardial fibrosis in the setting of diabetic cardiomyopathy. J Am Coll Cardiol 2006, 47(4):693-700.
• [6]Tanaka K, Honda M, Takabatake T: Redox regulation of MAPK pathways and cardiac hypertrophy in adult rat cardiac myocyte. J Am Coll Cardiol 2001, 37(2):676-685.
• [7]Cailleret M, Amadou A, Andrieu-Abadie N, Nawrocki A, Adamy C, Ait-Mamar B, et al.: N-acetylcysteine prevents the deleterious effect of tumor necrosis factor-(alpha) on calcium transients and contraction in adult rat cardiomyocytes. Circulation 2004, 109(3):406-411.
• [8]Fiordaliso F, Bianchi R, Staszewsky L, Cuccovillo I, Doni M, Laragione T, et al.: Antioxidant treatment attenuates hyperglycemia-induced cardiomyocyte death in rats. Mol Cell Cardiol 2004, 37(5):959-968.
• [9]Luo M, Guan X, Luczak ED, Lang D, Kutschke W, Gao Z, et al.: Diabetes increases mortality after myocardial infarction by oxidizing CaMKII. J Clin Invest 2013, 123(3):1262-1274.
• [10]Tremoleda JL, Kerton A, Gsell W: Anaesthesia and physiological monitoring during in vivo imaging of laboratory rodents: considerations on experimental outcomes and animal welfare. EJNMMI Research 2012, 2(1):44. BioMed Central Full Text
• [11]Schiller NB, Shah PM, Crawford M, DeMaria A, Devereux R, Feigenbaum H, et al.: Recommendations for quantitation of the left ventricle by two-dimensional echocardiography. American Society of Echocardiography Committee on Standards, Subcommittee on Quantitation of Two-Dimensional Echocardiograms. J Am Soc Echocardiogr 1989, 2(5):358-367.
• [12]Du J, Liu J, Feng HZ, Hossain MM, Gobara N, Zhang C, et al.: Impaired relaxation is the main manifestation in transgenic mice expressing a restrictive cardiomyopathy mutation, R193H, in cardiac TnI. Am J Physiol Heart Circ Physiol 2008, 294(6):H2604-2613.
• [13]Connelly KA, Kelly DJ, Zhang Y, Prior DL, Martin J, Cox AJ, et al.: Functional, structural and molecular aspects of diastolic heart failure in the diabetic (mRen-2)27 rat. Cardiovasc Res 2007, 76(2):280-291.
• [14]Yong QC, Thomas CM, Seqqat R, Chandel N, Baker KM, Kumar R: Angiotensin type 1a receptor-deficient mice develop diabetes-induced cardiac dysfunction, which is prevented by renin-angiotensin system inhibitors. Cardiovasc Diabetol 2013, 12:169. BioMed Central Full Text
• [15]Frangogiannis NG, Dewald O, Xia Y, Ren G, Haudek S, Leucker T, et al.: Critical role of monocyte chemoattractant protein-1/CC chemokine ligand 2 in the pathogenesis of ischemic cardiomyopathy. Circulation 2007, 115(5):584-592.
• [16]Banerjee I, Fuseler JW, Price RL, Borg TK, Baudino TA: Determination of cell types and numbers during cardiac development in the neonatal and adult rat and mouse. Am J Physiol Heart Circ Physiol 2007, 293(3):H1883-1891.
• [17]Xu YJ, Tappia PS, Neki NS, Dhalla NS: Prevention of diabetes-induced cardiovascular complications upon treatment with antioxidants. Heart Fail Rev 2014, 19(1):113-121.
• [18]Dhalla NS, Rangi S, Zieroth S, Xu YJ: Alterations in sarcoplasmic reticulum and mitochondrial functions in diabetic cardiomyopathy. Exp Clin Cardiol 2012, 17(3):115-120.
• [19]Ceriello A, Motz E: Is oxidative stress the pathogenic mechanism underlying insulin resistance, diabetes, and cardiovascular disease? The common soil hypothesis revisited. Arterioscler Thromb Vasc Biol 2004, 24(5):816-823.
• [20]Abdollahi M, Ranjbar A, Shadnia S, Nikfar S, Rezaie A: Pesticides and oxidative stress: a review. Med Sci Monit 2004, 10(6):RA141-147.
• [21]Shih CC, Wu YW, Lin WC: Antihyperglycaemic and anti-oxidant properties of Anoectochilus formosanus in diabetic rats. Clin Exp Pharmacol Physiol 2002, 29(8):684-688.
• [22]Naziroglu M, Butterworth PJ: Protective effects of moderate exercise with dietary vitamin C and E on blood antioxidative defense mechanism in rats with streptozotocin-induced diabetes. Can J Appl Physiol 2005, 30(2):172-185.
• [23]Shaw A, Doherty MK, Mutch NJ, MacRury SM, Megson IL: Endothelial cell oxidative stress in diabetes: a key driver of cardiovascular complications? Biochem Soc Trans 2014, 42(4):928-933.
• [24]Sampson N, Koziel R, Zenzmaier C, Bubendorf L, Plas E, Jansen-Durr P, et al.: ROS signaling by NOX4 drives fibroblast-to-myofibroblast differentiation in the diseased prostatic stroma. Mol Endocrinol 2011, 25(3):503-515.