Puerarin mediates hepatoprotection against CCl4-induced hepatic fibrosis rats through attenuation of inflammation response and amelioration of metabolic function

doi:10.1016/j.fct.2012.10.059

Food and Chemical Toxicology

Volume 52, February 2013, Pages 69-75

https://doi.org/10.1016/j.fct.2012.10.059 Get rights and content

Abstract

This study was designed to evaluate the potential effects of puerarin (PR), an effective isoflavonoid compound purified from Pueraria lobata, in treating hepatic fibrosis (HF) rats induced by carbon tetrachloride (CCl₄, 2 mL kg⁻¹ d⁻¹). Compared to model control, PR treatment effectively lowered the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (Alb), total protein (TP) in HF rats. Masson stained analysis showed that the condition of HF rats was mitigated. Meanwhile, the tumor necrosis factor alpha (TNF-α), nuclear factor-kappa B (NF-κB) expressions were significantly down-regulated at protein level by PR intervention. Additionally, the activity of superoxide dismutase (SOD) was elevated, while the content of malondialdehyde (MDA) was lessened in liver tissue. As revealed by immunohistochemistry assay, PR therapy resulted in reduced production of transforming growth factor-βl (TGF-βl). Moreover, it also was attributed to decreased mRNA level of inducible nitric oxide synthase (iNOS) using RT-PCR analysis. These findings demonstrate that puerarin successfully reverses hepatotoxicity in CCl₄-induced HF rats via the underlying mechanisms of regulating serum enzymes and attenuating TNF-α/NF-κB pathway for anti-inflammation response, as well as improving metabolic function in liver tissue.

Highlights

► Puerarin treatment is a candidate of anti-fibrosis. ► Puerarin effectively regulates serum enzymes. ► Puerarin attenuates inflammation response in liver tissue. ► Puerarin ameliorates metabolic function of liver. ► The effects of puerarin are beneficial of hepatoprotection.

Introduction

Chronic liver diseases progressively worsening will trigger hepatic fibrosis (HF) caused by the excessive accumulation of extracellular matrix (ECM), especially collagen protein. Aggravated HF may be characterized by necrosis, collapse and scar formation, eventually resulting in fatal cirrhosis (Bataller and Brenner, 2005, Iwaisako et al., 2012). Numerous evidences now has been proved to recognize hepatic stellate cells (HSCs) as the main matrix-producing cells involved in the HF development (Moreira, 2007). To date, there is no cure for the HF, but the conditions can be managed or controlled. Therefore, available methods are required to develop novel therapeutic strategies for anti-fibrotic treatment (Li and Friedman, 1999, Kisseleva and Brenner, 2011).

Pueraria lobata plant, a common herbal use in Chinese for centuries, has traditionally been applied as a remedy for alcohol alcoholism and the reward circuits in the brain (Keung and Vallee, 1998, McGregor, 2007, Li et al., 2010). Additionally, puerarin, one of the active components of P. lobata, has been reported to counteract alcohol intoxication achieved success in China (Benlhabib et al., 2004, Zhao et al., 2010). Meanwhile, it functions as the numerous pharmacological activities such as blood pressure normalization, antioxidant roles, and heart disease prevention and neurodegeneration control, as well as biological hepatoprotective effect (Xie et al., 2003, Zhu et al., 2010, Hwang et al., 2007). Even so, previously limited investigations are available about the effects of PR on CCl₄-induced cytotoxicity in rats. Hence, in present study, HF rat was used as an animal model to assess the anti-hepatofibrosis effects of PR against CCl₄-induced hepatotoxicity.

Section snippets

Materials

The puerarin preparation was provided from Guangxi Medical University of department of pharmaceutical chemistry (Nanning, China). Reagent grade of CCl₄ was obtained from Chengdu Kelong Chemical Reagent Factory (Chengdu, China). The sources for other required materials are labeled as below.

Animals and drug administration

Health male Wistar rats weighing 200 ± 20 g, SPF, were purchased from the Medical Laboratory Animal Center of Guangxi Medical University, China (Certificate No. SCXK-Gui-2009-0002). The research was conducted

Animal vital signs and liver index

After 60 days of administration, there were no death cases in normal control group, while ten deaths occurred in model control group (CCl₄-induced HF rats), as well as four, seven, five and two deaths happened in the low-, medium- and high-dose PR, colchicine treated groups, respectively. In addition, apathetic condition with emaciation and obvious body weight loss in rats were mostly exhibited in the model control group, and the therapies groups gradually reversed those symptoms in HF rats (

Discussion

Currently, an oriented tendency of hepatofibrosis management is urgent to discover effective compounds involved in low-toxicity. And pinpointed the hepatic fibrosis develops and discovered effective therapeutical strategy will pave the way for treatments to prevent fibrosis (Mills et al., 2005, Dossey, 2010, Teichert and Olivera, 2010). In our present study, the encouraging findings have indicated that puerarin (PR), a nature product isolated and purified from P. lobata, has beneficial effects

Conclusions

Taken together, our promising results indicate that puerarin may serve as a candidate with high potential to manage hepatofibrosis via its effective amelioration of metabolic function in liver. The effects of puerarin are probably through regulating serum enzymes and blocking TNF-α/NF-κB pathway for anti-inflammation response, as well as neutralizing cytotoxicity in liver tissue.

Conflict of Interest

The authors declare no conflicts of interest and are responsible for the contents of this study.

Acknowledgments

We are grateful to Prof. Jiaquan Li and Prof. Haiyuan Xie for their excellent technical supports.

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¹: First co-author, they contributed equally.

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Puerarin mediates hepatoprotection against CCl4-induced hepatic fibrosis rats through attenuation of inflammation response and amelioration of metabolic function

Abstract

Highlights

Introduction

Section snippets

Materials

Animals and drug administration

Animal vital signs and liver index

Discussion

Conclusions

Conflict of Interest

Acknowledgments

Toxicol. Appl. Pharmacol.

Gastroenterology

J. Hepatol.

Mol. Aspects Med.

Phytochemistry

Best Pract. Res. Clin. Gastroenterol.

Best Pract. Res. Clin. Gastroenterol.

Eur. J. Pharmacol.

J. Ethnopharmacol.

Alcohol

Toxicol. Appl. Pharmacol.

Int. J. Biochem. Cell Biol.

Nuclear receptors, inflammation, and liver disease: insights for cholestatic and fatty liver diseases

Clin. Pharmacol. Ther.

Liver fibrosis

J. Clin. Invest.

Effects of purified puerarin on voluntary alcohol intake and alcohol withdrawal symptoms in P rats receiving free access to water and alcohol

J. Med. Food

The natural history of nonalcoholic fatty liver disease with advanced fibrosis or cirrhosis: an international collaborative study

Hepatology

Mechanisms of hepatic toxicity. I. TNF-induced liver injury

Am. J. Physiol.

The NF-kappaB regulatory network

Cardiovasc. Toxicol.

TNF-R1 signaling: a beautiful pathway

Science

The SODyssey: superoxide dismutases from biochemistry, through proteomics, to oxidative stress, aging and nutraceuticals

Expert Rev. Proteomics

Insects and their chemical weaponry: new potential for drug discovery

Nat. Prod. Rep.

Betaine or taurine administration prevents fibrosis and lipid peroxidation induced by rat liver by ethanol plus carbon tetrachloride intoxication

Amino Acids

Liver enzyme alteration: a guide for clinicians

Can. Med. Assoc. J.

Hepatic fibrosis and cirrhosis after chronic administration of alcohol and “low-dose” carbon tetrachloride vapor in the rat

Hepatology

Redox regulation of tumor necrosis factor signaling

Antioxid. Redox Signaling

Puerarin mediates hepatoprotection against CCl₄-induced hepatic fibrosis rats through attenuation of inflammation response and amelioration of metabolic function