Chlorogenic acid ameliorates alcohol-induced liver injuries through scavenging reactive oxygen species

doi:10.1016/j.biochi.2018.05.008

Biochimie

Volume 150, July 2018, Pages 131-138

https://doi.org/10.1016/j.biochi.2018.05.008 Get rights and content

Highlights

•
Alcoholic liver disease (ALD) is associated with oxidative stress.
•
Chlorogenic acid (CGA) plays a role in preventing ALD through its antioxidant properties.
•
CGA reduced alcohol-induced-steatosis, apoptotic cell death, and fibrosis in liver.

Abstract

The key role of oxidative stress in alcoholic liver disease (ALD) has been established by the large body of evidence from previous studies. Excessive consumption of ethanol induces the production of a variety of reactive oxygen species (ROS) in the liver, such as superoxide, H₂O₂, and hydroxyl radical. These products activate oxidant-sensitive signaling cascades and modulators of apoptosis. Because ROS accumulation is closely related to ALD, a number of studies have investigated the benefits of antioxidants. Recent studies demonstrated that polyphenol chlorogenic acid (CGA) has antioxidant properties and health benefits, such as reduction of relative risk of cardiovascular diseases and hepatoprotective effects against acetaminophen toxicity. However, the protective effects of CGA against ALD have not been studied in detail. We hypothesize that CGA plays a role in preventing ALD through its antioxidant properties. In this study, we investigated the protective effects of CGA against liver injuries in vivo. Reduced alcohol-induced-steatosis, apoptotic cell death, and fibrosis due to reduced levels of oxidative stress were observed. These findings suggest that CGA treatment can be an effective approach to attenuate ALD through the suppression of oxidative stress.

Graphical abstract

Introduction

Alcoholic liver disease (ALD) is a major cause of death due to liver injuries, including steatosis, steatohepatitis, fibrosis, and cirrhosis [1]. Many studies on pathogenesis of ALD have focused on oxidative stress and glutathione depletion, abnormal methionine metabolism, malnutrition, and endotoxin production [2], and accumulating evidence suggests that oxidative stress plays a key role in ALD [3].

Ethanol induces oxidative stress through various routes. These include mitochondrial damage, induction of CYP2E1, ethanol-induced activation of Kupffer cells, and depletion of mitochondrial and cytosolic glutathione. These routes are not mutually exclusive, and lead to the generation of reactive oxygen species (ROS) and induction of oxidative stress [4]. Therefore, excessive consumption of ethanol induces the production of a variety of ROS, such as superoxide, H₂O₂, and hydroxyl radical in liver cells [5]. These products cause cellular injury through alteration of biomolecules or interference with the signaling cascades. Previous studies have shown the induction of hepatic apoptosis via mitochondrial dysfunction and liver injury through alcohol-induced ROS generated during ethanol metabolism [6]. Furthermore, some investigators demonstrated that alcohol consumption stimulates the inflammatory responses by activating NF-κB and MAPK in the liver cells [7], and these alterations are caused as a result of ROS production [8,9].

Because high oxidative stress is closely related with ALD, a number of studies have investigated the benefits of antioxidants to protect the liver from ALD. Silymarin, vitamin E, and N-acetylcysteine are representative antioxidants that have been studied in relation to liver health [[10], [11], [12]]. Chlorogenic acid (CGA) is the ester of caffeic acid and quinic acid [13], and many dietary plants such as coffee, tea, and some fruits and vegetables contain CGA. Recent studies have shown that the consumption of CGA has health benefits, such as reduction in the relative risk of cardiovascular diseases, diabetes type 2, and Alzheimer's disease. It also has anti-bacterial and anti-inflammatory properties, and exhibits hepatoprotective effects against acetaminophen toxicity [14,15]. However, the antioxidant property and protective effect of CGA with respect to ALD have not been studied. We hypothesized that CGA plays a role in preventing ALD because of its antioxidant properties. We demonstrated the protective effect of CGA against liver injuries in an ALD model. Reduced alcohol-induced steatosis, apoptotic cell death, and fibrosis due to reduced levels of oxidative stress were observed. These findings suggest that CGA can be used to attenuate ALD through the suppression of oxidative stress.

Section snippets

Materials

Oil Red O solution, CGA, JC-1, rhodamine 123, and Serum Triglyceride Determination Kit were purchased from Sigma (St. Louis, MO, USA). MitoTracker and mitoSox were purchased from Invitrogen (Eugene, OR, USA). In Situ Cell Death Detection Kit was from Roche (Basel, Switzerland). Alanine transaminase (ALT) assay kit and aspartate transaminase (AST) assay kit were purchased from Abnova (Taipei, Taiwan). Antibodies were purchased as follows: β-Actin, JNK and SREBP1 (Santa Cruz Biotechnology, Santa

Result and discussion

To observe the protective effect of CGA against alcohol-induced liver injuries, the serum levels of the biomarkers, ALT and AST, were determined. As shown in Fig. 1A, ALT and AST levels were higher in the ethanol-treated group than in the vehicle group. When mice were injected with 10, 20, or 40 mg/kg of CGA, the dose-dependent attenuation of ALT and AST activities was observed in the CGA-treated sera compared to that in the alcohol only group. The subsequent experiments were conducted with

Acknowledgement

This work was supported by the National Research Foundation of Korea (NRF) grant (NRF-2015R1A4A1042271) funded by the Korea Government (MSIP).

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Aflatoxin B1 (AFB1) is widely distributed in crops and feeds, and ingestion of AFB1-contaminated crops is harmful to human/animal health. This study was designed to investigate hepatoprotective effects of chlorogenic acid (CGA), due to its excellent antioxidant and anti-inflammatory activities, on mice exposed to AFB1. Male Kunming mice were orally fed with CGA prior to daily AFB1 exposure for 18 consecutive days. The results showed that CGA treatment reduced the serum activity of aspartate aminotransferase, hepatic malondialdehyde content and pro-inflammatory cytokines synthesis, prevented histopathological changes of the liver, increased hepatic glutathione level, catalase activity and IL10 mRNA expression in mice subjected to AFB1. Taken together, CGA exerted the protective effect on AFB1-induced hepatic damage by modulating redox status and inflammation, suggesting that CGA may be a candidate compound for the treatment of aflatoxicosis.
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Chlorogenic acid (CGA) is a phenylpropanoid compound in chrysanthemum, which has a variety of biological activities. Nevertheless, CGA has not yet been studied concerning oxidative stress in hepatocytes. Our research aims to investigate whether chlorogenic acid can improve hepatic oxidative stress, and further illustrated its detailed mechanism. The results of free radicals measurement and Western blot showed that CGA suppressed the activation of JNK (c-Jun NH2-teminal kinase) signaling pathway by ameliorating endoplasmic reticulum stress in H₂O₂-treated hepatocytes, thereby eliminating excess intracellular free radicals. In addition, through RT-qPCR, we found that miR-199a-5p down-regulated in H₂O₂-induced L02 cells, which was restored by CGA. By inhibiting the expression of miR-199a-5p to promote the transcription of GRP78 gene, the antioxidant capacity of CGA was greatly weakened in hepatocytes. Our findings suggest that CGA treatment alleviated the H₂O₂-induced oxidative stress by targeting miR-199a-5p, thereby ameliorating ER stress and inhibiting JNK phosphorylation for antioxidant capacity enhancement. Based on our available results, CGA may be an alternative compound to mitigate hepatic oxidative damage through the miR-199a-5p/GPR78 axis.
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Mice in colitis+periodontitis group displayed increased periodontal inflammation and alveolar bone loss when compared with the mice of periodontitis group, suggesting colitis aggravated periodontitis. Metabolomics analysis combined with enrichment analysis showed that colitis significantly (P＜0.05) altered the content of compounds associated with five metabolic pathways (e.g. fatty acid biosynthesis) of periodontitis mice. Notably, colitis significantly reduced the level of serum metabolites that inhibited the formation of osteoclasts (e.g. oleic acid) or anti-inflammatory metabolites (e.g. palmitoleic acid, palmitelaidic acid and chlorogenic acid) of periodontitis mice.
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Ameliorative effects of chlorogenic acid on alcoholic liver injury in mice via gut microbiota informatics
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Citation Excerpt :
These results indicated that ingestion of CGA may inhibit alcohol-induced liver damage, and we thus continue to explore its effects on hepatic biochemical indices and pathological alterations. Oxidative stress is one of the key points that cannot be ignored in the development of ALD (Kim et al., 2018). When alcohol is metabolized in the liver, a large number of free radicals are generated, destroying the previously balanced redox system (Ceni et al., 2014).
Chlorogenic acid (CGA) is a functional phenolic acid widely used in food and medicine-related fields. It has been proved to be effective in the treatment of alcoholic liver disease (ALD). However, the exact mechanism by which CGA prevents ALD, especially from the crosstalk between gut and liver, has not been previously reported. This work was aimed to explore the protective effects of CGA against ALD and its relationships to gut-liver axis abnormalities. Experimental results showed the increased (p < 0.05) serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), low density lipoprotein (LDL), total cholesterol (TC) and triglyceride (TG) levels of mice fed with ethanol were ameliorated by supplementing with CGA. Moreover, CGA promoted the production of n-butyric acid by nearly 3 times (1.78 vs 0.62 nM, p < 0.01), a short-chain fatty acid that helps maintain the integrity of the intestinal barrier. Furthermore, CGA alleviated microbial dysbiosis, evidenced by the increased relative abundances of beneficial bacteria Muribaculaceae, Bacteroides, Alloprevotella, and Parabacteroides, and decreased that of opportunistic pathogens Eubacterium_nodatum, Eubacterium_ruminantium, and Anaerotruncus. Correlation analysis further elucidated the microbiota altered after CGA intervention was positively correlated with short-chain fatty acids and antioxidant indexes, while negatively correlated with inflammatory cytokines. In summary, these findings suggested the hepatoprotective effect of CGA was ascribed to the modulation of gut-liver axis homeostasis.
Chlorogenic acid-water complexes in chlorogenic acid containing food products
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Citation Excerpt :
The adoption of IUPAC nomenclature of CQA isomers will allow the reader to easily relate the results of this work to the results of previous works (Dawidowicz and Typek, 2010; Dawidowicz and Typek, 2011, 2012, 2014, 2015). The results of epidemiological research suggest that consumption of food, beverages (like tea or coffee) and diet supplements containing 5-CQA may help to prevent several chronic diseases, including type 2 diabetes mellitus, Parkinson's disease and liver diseases (cirrhosis and hepatocellular carcinoma) (Yan et al., 2017; Kim et al., 2018; Cao et al., 2019; Fukutomi et al., 2021; Araújo et al., 2021). These findings are responsible for the growing interest in the natural occurrence of CQAs and their properties.
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Research paperChlorogenic acid ameliorates alcohol-induced liver injuries through scavenging reactive oxygen species

Highlights

Abstract

Graphical abstract

Introduction

Section snippets

Materials

Result and discussion

Acknowledgement

Gastroenterology

Alcohol

Gastroenterology

J. Nutr.

J. Nutr. Biochem.

Anal. Biochem.

Methods Enzymol.

Free Radic. Biol. Med.

Biochimie

Hepatology

Canc. Lett.

Life Sci.

Alcoholic liver disease: pathogenesis and new targets for therapy

Nat. Rev. Gastroenterol. Hepatol.

Role of oxidative stress in alcohol-induced liver injury

Arch. Toxicol.

Alcohol and oxidative liver injury

Hepatology

Oxidative stress and signal transduction pathways in alcoholic liver disease

Alcohol Clin. Exp. Res.

Mechanisms of Liver Injury. I. TNF-alpha-induced liver injury: role of IKK, JNK, and ROS pathways

Am. J. Physiol. Gastrointest. Liver Physiol.

Research paper
Chlorogenic acid ameliorates alcohol-induced liver injuries through scavenging reactive oxygen species