Herbal Supplement Ameliorates Cardiac Hypertrophy in Rats with Ccl4-Induced Liver Cirrhosis

Hindawi Evidence-Based Complementary and Alternative Medicine Volume 2017, Article ID 5276749, 1 page https://doi.org/10.1155/2017/5276749 Retraction Retracted: Herbal Supplement Ameliorates Cardiac Hypertrophy in Rats with CCl4-Induced Liver Cirrhosis Evidence-Based Complementary and Alternative Medicine Received 8 August 2017; Accepted 8 August 2017; Published 22 November 2017 Copyright © 2017 Evidence-Based Complementary and Alternative Medicine. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Evidence-Based Complementary and Alternative Medicine has retracted the article titled “Herbal Supplement Amelio- ratesCardiacHypertrophyinRatswithCCl4-Induced Liver Cirrhosis” [1]. The article was found to contain duplicated images in Figure 4(a), in which the IL6 blots are identical to the p-ERK5 blots. The authors provided a corrected figure, with replacements for p-ERK5, which is available as Supplementary Materials. However, they could not provide the underlying blots. References [1] P.-C. Li, Y.-W. Chiu, Y.-M. Lin et al., “Herbal supplement ameliorates cardiac hypertrophy in rats with CCl4-induced liver cirrhosis,” Evidence-Based Complementary and Alternative Medicine, vol. 2012, Article ID 139045, 9 pages, 2012. Hindawi Publishing Corporation Evidence-Based Complementary and Alternative Medicine Volume 2012, Article ID 139045, 9 pages doi:10.1155/2012/139045 Research Article Herbal Supplement Ameliorates Cardiac Hypertrophy in Rats with CCl4-Induced Liver Cirrhosis Ping-Chun Li,1, 2 Yung-Wei Chiu,3, 4 Yueh-Min Lin,5 Cecilia Hsuan Day,6 Guang-Yuh Hwang,2 Peiying Pai,7 Fuu-Jen Tsai,8 Chang-Hai Tsai,9 Yu-Chun Kuo,10 Hsiao-Chuan Chang,11 Jer-Yuh Liu,12, 13 and Chih-Yang Huang8, 10, 14 1Division of Cardiovascular Surgery, China Medical University Hospital, Taichung 40402, Taiwan 2Department of Life Science, Tunghai University, Taichung 40704, Taiwan 3Emergency Department and Center of Hyperbaric Oxygen Therapy, Tungs’ Taichung MetroHarbor Hospital, Taichung 43503, Taiwan 4Institute of Medicine, Chung Shan Medical University, Taichung 40201, Taiwan 5Department of Pathology, Changhua Christian Hospital, Changhua 50006, Taiwan 6Department of Nursing, MeiHo University, Pingtung 91202, Taiwan 7Division of Cardiology, China Medical University Hospital, Taichung 40402, Taiwan 8Graduate Institute of Chinese Medical Science, China Medical University, Taichung 40402, Taiwan 9Department of Healthcare Administration, Asia University, Taichung 41354, Taiwan 10Graduate Institute of Basic Medical Science, China Medical University, Taichung 40402, Taiwan 11Department of Biotechnology, Asia University, Taichung 41354, Taiwan 12Center for Molecular Medicine, China Medical University Hospital, Taichung 40402, Taiwan 13Graduate Institute of Cancer Biology, China Medical University, Taichung 40402, Taiwan 14Department of Health and Nutrition Biotechnology, Asia University, Taichung 41354, Taiwan Correspondence should be addressed to Jer-Yuh Liu, [email protected] and Chih-Yang Huang, [email protected] Received 31 May 2012; Revised 31 July 2012; Accepted 7 August 2012 Academic Editor: Y. Ohta Copyright © 2012 Ping-Chun Li et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. We used the carbon tetrachloride (CCl 4) induced liver cirrhosis model to test the molecular mechanism of action involved in cirrhosis-associated cardiac hypertrophy and the effectiveness of Ocimum gratissimum extract (OGE) and silymarin against cardiac hypertrophy. We treated male wistar rats with CCl4 and either OGE (0.02 g/kg B.W. or 0.04 g/kg B.W.) or silymarin (0.2 g/kg B.W.). Cardiac eccentric hypertrophy was induced by CCl4 along with cirrhosis and increased expression of cardiac hypertrophy related genes NFAT, TAGA4, and NBP, and the interleukin-6 (IL-6) signaling pathway related genes MEK5, ERK5, JAK, and STAT3. OGE or silymarin co-treatment attenuated CCl4-induced cardiac abnormalities, and lowered expression of genes which were elevated by this hepatotoxin. Our results suggest that the IL-6 signaling pathway may be related to CCl4-induced cardiac hypertrophy. OGE and silymarin were able to lower liver fibrosis, which reduces the chance of cardiac hypertrophy perhaps by lowering the expressions of IL-6 signaling pathway related genes. We conclude that treatment of cirrhosis using herbal supplements is a viable optionRETRACTED for protecting cardiac tissues against cirrhosis-related cardiac hypertrophy. 1. Introduction patients with cirrhosis and potential clinical implications have long been known [5], little is understood regarding Patients with advanced cirrhosis have consistently been the molecular mechanism of action involved in cirrhosis- diagnosed with cardiac dysfunction under the condition associated alteration in cardiac structure and function, of hyperdynamic circulation [1]. Increased cardiac output especially cardiac hypertrophy. and reduced systemic vascular resistance are both signs Cirrhosis is known as a possible cause of portal vein of this condition [2–4]. Although cardiac dysfunction in constriction which may induce the activation of vasopressin, 2 Evidence-Based Complementary and Alternative Medicine angiotensin II (Ang II), and the sympathetic nervous system intraperitoneally with CCl4 (8% CCl4/corn oil, 1 mL/kg body [6]. Cardiac hypertrophy is induced by such direct mechan- weight (BW) twice a week, Monday and Thursday) for 8 ical wall stress as well as paracrine/autocrine factors such as weeks, as described by Hernandez-Muoz´ et al. [23], with Ang II, which in turn activates specific signaling pathways, some modifications. At the same time, the rats were treated for instance, mitogen-activated protein kinases (MAPKs) with various dosages of OGE (0–0.04 g/kg BW), or silymarin and calcineurin. These can cause cardiac hypertrophy and orally (0.2 g/kg BW, four times a week, Tuesday, Wednesday, increase of related gene expressions, such as proto-oncogenes Friday, and Saturday) [24, 25]. The control rats were treated c-Fos and c-JUN, genes which encode atrial natriuretic with corn oil (1 mL/kg BW) and fed a normal diet. At the peptide (ANP) and B-type natriuretic peptide (BNP), and end of the experiment, blood and heart were immediately structural genes β-myosin heavy chain (β-MHC) and skeletal obtained after the animals were sacrificed. α-actin [7]. Ang II is associated with increased plasma levels of proinflammatory cytokines such as interleukin-6 (IL-6) 2.3. Histological Examinations. The heart was fixed in 10% [8], which is an effective stimulator of the Janus kinase/signal formalin, processed using routine histology procedures, transducer and activator of transcription (JAK/STAT) path- embedded in paraffin, cut in 5 μm sections, and mounted on way in cardiac hypertrophy [7]. However, the role of a slide. The samples were stained with hematoxylin and eosin these protein markers and transcriptional factors in cardiac for histopathological examination. hypertrophy and remodeling in vivo has not been examined in cirrhosis-associated hypertrophy. 2.4. Preparation of Tissue Extract. Allprocedureswereper- ◦ Carbon tetrachloride (CCl4) is frequently used to induce formed at 4 C. The heart samples were lysed by 30 strokes experimental cirrhosis in rats [9]. This model has recently using a Kontes homogenizer at a ratio of 100 mg tissue/1 been used to investigate the role of lipophilic bile acids mL lysis buffer. The lysis buffer consisted of 50 mM Tris- and examine cardiac gene expression profiles in cirrhotic HCl (pH 7.4), 2 mM EDTA, 2 mM EGTA, 150 mM NaCl, cardiomyopathy [10, 11]. Silymarin, a standardized extract 1mMPMSF,10μg/mL leupeptin, 1 mM sodium orthovana- of the milk thistle (Silybum marianum L. Gaertner), contains date, 1% (v/v) 2-mercaptoethanol, 1% (v/v) Nonidet P40, three biochemicals: silybin, silydianin, and silychristin and and 0.3% sodium deoxycholate. These homogenates were has a long tradition as a herbal remedy [12]. Ocimum centrifuged at 100,000 g for 1 h at 4◦C. The supernatant was gratissimum extract(OGE),acommonlyusedherbinfolk stored at −70◦C for Western blot assay. medicine, is rich in antioxidants and possesses many ther- apeutic functions [13–21]. Both herbal extracts have been 2.5. Electrophoresis and Western Blot. Tissue extract sam- shown using the CCl4 model to inhibit liver cirrhosis [22]. ples were prepared as described above. Sodium do deco Therefore the motive for this experiment is to use the CCl4- sulfate-polyacrylamide gel electrophoresis is carried out as induced liver cirrhosis model to understand the molecular described by Laemmli [26] using 10% polyacrylamide gels. mechanism of action involved in cirrhosis-associated cardiac After samples are electrophoresed at 140 V for 3.5 h, the hypertrophy, as well as to test effectiveness of silymarin and gels are equilibrated for 15 min in 25 mM Tris-HCl, pH OGE against cardiac damage and hypertrophy. 8.3, containing 192 mM glycine and 20% (v/v) methanol. Electrophoresed proteins are transferred to nitrocellulose 2. Materials and Methods paper (Amersham, Hybond-C Extra Supported, 0.45 Micro) using Hoefer Scientific Instruments Transpher Units at 2.1. Preparation of OGE. Leaves of Ocimum gratissimum 100 mA for 14 h. The nitrocellulose paper was incubated

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