effects of da-5513 on alcohol metabolism and alcoholic ...effects of da-5513 on serum alcohol and...

49

Lab Anim Res 2018: 34(2), 49-57

https://doi.org/10.5625/lar.2018.34.2.49

ISSN 1738-6055 (Print)

ISSN 2233-7660 (Online)

Effects of DA-5513 on alcohol metabolism andalcoholic fatty liver in rats

Jae Young Yu1,#, Hanh Thuy Nguyen2,3,#, Chul Soon Yong2, Hyoung Geun Park1, Joon Ho Jun1, Jong Oh Kim2,*1Department of Formulation Development, Dong-A Pharmaceutical Co. Ltd., Yongin, Korea

2College of Pharmacy, Yeungnam University, Gyeongsan, Korea3National Institute of Pharmaceutical Technology, Hanoi University of Pharmacy, Hanoi, Vietnam

Hangover is characterized by a number of unpleasant physical and mental symptoms that occur afterheavy alcohol drinking. In addition, consistently excessive alcohol intake is considered as a major reasoncauses liver disease. The present study investigated the in vivo effects of DA-5513 (Morning care® KangHwang) on biological parameters relevant to hangover relief and alcoholic fatty liver. Blood alcohol andacetaldehyde concentrations were determined in rats administered a single dose of alcohol and treatedwith DA-5513 or commercially available hangover relief beverages (Yeomyung® and Ukon®). The effectsof DA-5513 on alcoholic fatty liver were also determined in rats fed alcohol-containing Lieber-DeCarlidiets for 4 weeks. Serum liver function markers (aspartate and alanine aminotransferase activities) andserum/liver lipid levels were assessed. Blood alcohol and acetaldehyde concentrations were lower in thegroups treated with DA-5513 or Yeomyung®, as compared with control rats. However, Ukon® did notproduce any significant effects on these parameters. Treatment with DA-5513 significantly reduced serumaspartate and alanine aminotransferase activities and markedly reduced serum cholesterol andtriglyceride levels, as compared with control rats. Histological observations using Oil Red O stainingfound that DA-5513 delayed the development of alcoholic fatty liver by reversing hepatic fataccumulation. These findings suggest that DA-5513 could have a beneficial effect on alcohol-inducedhangovers and has the potential to ameliorate alcoholic fatty liver.

Keywords: Morning care®, hangover, acetaldehyde, alcohol-induced fatty liver, hepatic triglyceride

Received 9 March 2018; Revised version received 19 April 2018; Accepted 25 April 2018

Alcoholic liver disease has been demonstrated to be a

major cause of morbidity and mortality worldwide in

individuals with consistently excessive alcohol intake

[1]. Hangover is characterized by a number of unpleasant

physical and mental symptoms that occur after heavy

alcohol drinking [2,3]. Alcohol is initially oxidized to

acetaldehyde by the alcohol dehydrogenase (ADH)

enzyme; this is subsequently converted to acetate by

aldehyde dehydrogenase (ALDH) in the liver [2,3].

Acetaldehyde is much more toxic than ethanol and this

metabolite may cause the physical symptoms of hangover

such as fatigue, headache, increased sensitivity to light

and sound, redness of the eyes, muscle aches, and thirst

[3]. Furthermore, long-term consumption of alcohol in

large quantities may cause chronic liver diseases and

hepatic steatosis (alcoholic fatty liver), which is defined

as excess lipid accumulation in the cytoplasm of hepatocytes;

this is regarded as a significant risk factor for hepatic

fibrosis and cirrhosis [4]. Thus, reduction of alcohol-

induced hepatic fat accumulation may block or delay the

progression of steatosis to advanced stages of alcoholic

liver disease.

Multiple mechanisms contribute to the pathogenesis of

alcoholic hepatic steatosis, including increased de novo

#These authors contributed equally to this work.

*Corresponding author: Jong Oh Kim, 214-1, Dae-dong, College of Pharmacy, Yeungnam University, Gyeongsan, 712-749, KoreaTel: +82-53-810-2813; Fax: +82-53-810-4654; E-mail: [email protected]

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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50 Jae Young Yu et al.

Lab Anim Res | June, 2018 | Vol. 34, No. 2

hepatic lipogenesis, impaired mitochondrial fatty acid β-

oxidation, and reduced export of very low-density lipoprotein

[5]. However, the complex mechanisms involved in

alcoholic fatty liver formation have been debated in the

literature [6]. Accumulating evidence has suggested that

adipose tissue dysfunction might impact on hepatic lipid

metabolism [7,8]. A direct link between adipose triglyceride

(TG) loss and hepatic TG gain was revealed using

deuterium-labeled TG in alcohol-fed mice [9]. Dysregulation

of lipid homeostasis was also evidenced in clinical

studies, which identified a lower fat mass during the

development of alcoholic fatty liver [10]. Adipose tissue

plays a crucial role as a major metabolic buffering system

in lipid homeostasis [11,12], and modification of alcohol-

induced dysfunction in this tissue might therefore provide

an important target for the development of functional

foods to prevent alcoholic fatty liver [13]. Many treatments

have been reported to prevent and/or reduce the severity

of hangover and alcoholic fatty liver symptoms, including

innumerable folk remedies and recommendations.

Yeomyung® (YM, Glami, Gwangwon, Korea) is the

most commonly sold hangover recovery drink in Korea,

and Ukon® (UK, Enagic, Okinawa, Japan) is a turmeric

extract-based beverage that is marketed as an anti-

hangover drink and has enjoyed huge success in Japan.

In addition, various products have been released on the

basis that they decrease the accumulation of fat in the

liver, prevent liver damage, and alleviate alcohol-related

hangovers. However, the mechanism of action of these

products has not been revealed, and scientific research

into this area is therefore required. DA-5513 (Morning

care® Kang Hwang) was developed by the Dong-A

Pharm. Co. (Yongin, Korea) and has been approved by

the Korea Food and Drug Administration as an over-the-

counter treatment for hangover. The proprietary liquid

DA-5513 formulation was developed using seven herbal

extracts, including GMT-ALC-5L (fermented rice embryo

and bean extract), Curcuma longa L., Trapa japonica

Flerov., Silybum marianum L., Paullinia cupana Mart.,

Pueraria thunbergiana Benth., and honey. These natural

products have been used in traditional oriental medicines

to prevent alcohol-induced hangover and protect the

liver against diverse hepatotoxins such as ethanol, carbon

tetrachloride, antitubercular agents, and thioacetamide

[14-20].

The present study was therefore designed to investigate

the effect of DA-5513 on hangover relief, which would

be associated with the rapid elimination of alcohol and

acetaldehyde. Furthermore, a chronic ethanol-treated rat

model was developed to evaluate the ability of this

product to protect from alcoholic fatty liver.

Materials and Methods

Materials

DA-5513 was provided by the Dong-A Pharm. Co.

(Yongin, Korea), while Yeomyung® (YM, Glami,

Gwangwon, Korea) and Ukon® (UK, Enagic, Okinawa,

Japan) were purchased from the market. All other

chemicals used were of analytical grade.

Animals

Seven-week-old male Wistar rats were purchased

from Chung-Ang Lab Animal Inc. (Seoul, Korea) and

acclimatized to the laboratory setting (22.0±2.0oC, 12-h

light/dark cycles, 55%±5% humidity) with free access to

water and food (Samyang Co, Incheon, Korea) for a

week before the experiment. The experimental protocol

was approved by the Institutional Animal Care and Use

Committee of Dong-A Pharm. Co. (Yongin, Korea), and

all experimental procedures were conducted in compliance

with this company’s guidelines for the care and use of

laboratory animals.

Alcohol-induced hangover model

The rats (200-250 g) were randomly divided into four

groups (n=8 per group). Each group had a single oral

administration of the following test treatments: ethanol

(control group), and ethanol with either DA-5513, YM,

or UK. Then, after 0.5 h, 10 mL/kg of body weight was

administered. At 0.5, 1, 2, 4, and 6 h post-ethanol

administration, 0.2 mL blood was collected from each

rat and left at room temperature for 30 min before

centrifugation at 3,000 rpm for 15 min. The levels of

alcohol and acetaldehyde in the supernatant were

measured using kits for the detection of ethanol (Roche

Co., Darmstadt, Germany) and acetaldehyde (R-Biopharm,

Darmstadt, Germany). During the metabolism of ethanol

to acetaldehyde and acetate, nicotinamide adenine

dinucleotide (NAD+) is converted to NADH. Thus, the

concentration of NADH was determined by measuring

absorbance at 340 nm.

Alcoholic fatty liver model

Rats (220-230 g) were randomly divided into four

groups based on their body weight (n=10 per group):

Effects of DA-5513 on alcohol metabolism and alcoholic fatty liver 51


untreated control (CON), ethanol-treated control (ED),

and ethanol-treated with UK or DA-5513 groups. The

rats were fed a standard Lieber-DeCarli ethanol diet

(36% ethanol-derived calories) for 4 weeks [21,22]; pair-

fed control rats were administered dextran-maltose to

match the alcohol-derived calories in the ethanol diet.

DA-5513 or UK was introduced into the alcohol diet by

gradually mixing it with distilled water and feeding it at

the same time each day throughout the experiment. The

rats were sacrificed using ether anesthesia; blood samples

were centrifuged (1,500 g, 4oC, 10 min) to separate the

serum and stored at −80oC until analysis. The livers were

quickly removed and preserved in phosphate-buffered

formalin for histological examination. The rest of the

liver was frozen at −80oC prior to analysis of hepatic TG

levels.

Serum levels of alanine aminotransferase (ALT),

aspartate aminotransferase (AST), total cholesterol (T-

CHO), and TG were monitored by standard clinical

chemistry assays on an Automated Chemistry Analyzer

(Prestige 24I; Tokyo Boeki Medical System, Tokyo,

Japan). Total liver lipids were extracted from homogenates

prepared from 100 mg rat liver using chloroform:methanol

(2:1, v/v) [23]. The TG levels within total lipid samples

were determined enzymatically using a commercially

available enzymatic kit (Sigma Chem. Co.) according to

the manufacturer’s protocol.

Histological study

As proposed by Levene et al., Oil Red O can be used

to identify lipids and quantify hepatic steatosis. Several

protocols have been developed, including paraffin-

embedded sections and cryosections (or frozen sections)

[24,25]. In this study, frozen liver tissues were cut into

4-mm sections and affixed to microscope slides. Sections

were stained with Oil Red O solution buffer. Histo-

pathologic examinations of the liver sections were

conducted by a pathologist and were peer-reviewed. The

slides were dehydrated, dealcoholized, mounted utilizing

Canada balsam, and assessed for inflammation and tissue

damage utilizing an Olympus microscope (Olympus,

Tokyo, Japan) [26,27].

Statistical analysis

Data are given as the mean and standard deviation, and

inter-group differences were analyzed using one-way

ANOVA by Duncan’s method. Differences were considered

significant at a P-value <0.05, and very significant at a

P-value <0.01.

Results

Effects of DA-5513 on serum alcohol and acetaldehyde

concentrations

In order to determine the effects of DA-5513 on

hangovers, blood alcohol levels were investigated and

presented in Table 1. DA-5513, YM, or UK was

administered orally 0.5 h before 25% ethanol consumption,

and blood was collected 0.5, 1, 2, 4, and 6 h after alcohol

was administered. Administration of UK resulted in a

lower blood alcohol concentration than that observed in

the control group; however, this difference was not

statistically significant (P>0.05). On the other hand, DA-

5513 and YM produced a similar significant reduction of

the blood alcohol concentration at each of the time-

points examined, as compared with the control group

(Table 1).

The blood acetaldehyde concentration, which was

highest in the CON group, peaked 0.5 h after ethanol

consumption (Table 2). Rats treated with UK showed

lower blood acetaldehyde levels than control rats, but

these differences were not statistically significant. Both

DA-5513 and YM significantly reduced the acetaldehyde

levels in the blood at every time-point examined, as

Table 1. Blood alcohol concentrations (mg/mL) in rats

GroupTime (h)

0.5 1 2 4 6

Control 53.68±3.65 49.22±2.78 54.63±1.65 46.89±2.89 41.22±3.22

UK 49.66±2.16 50.55±3.22 49.33±4.12 43.22±5.13 38.55±5.32

YM 45.63±1.99* 43.56±2.35* 42.55±1.48* 33.99±2.99* 32.56±3.11*

DA-5513 43.66±3.55* 43.56±3.66* 42.31±2.33* 37.88±3.22* 31.47±2.98*

Control, ethanol-treated control; UK, treated with ethanol and UK; YM, treated with ethanol and YM; DA-5513, treated with ethanoland DA-5513.The data represent mean±standard deviation (n=8).*P<0.01 vs Control



compared with the control group. However, this effect

was more marked for DA-5513 than for YM throughout

the study. Taken together, these findings indicated that

DA-5512 provided more effective reduction of alcohol

and acetaldehyde levels than other products, suggesting

that it has the potential to act as a hangover relief

beverage.

Effects of DA-5513 on serum AST and ALT

All rats were fed the standard Lieber-DeCarli ethanol

diet for 4 weeks and hepatotoxicity was evaluated by

clinical chemistry. As seen in Figure 1A and 1B, the ED

group showed a marked increase in the levels of serum

AST and ALT (by approximately 1.3- and 1.4-fold,

respectively), as compared to the untreated control group

Table 2. Blood acetaldehyde concentrations (µg/mL) in rats

GroupTime (h)

0.5 1 2 4 6

Control 85.65±5.66 68.44±1.99 54.65±2.22 53.22±3.64 39.55±1.22

UK 80.66±7.19 60.55±6.88 48.66±6.59 47.55±5.23 35.66±4.66

YM 64.88±8.22* 55.89±5.69* 46.55±4.55* 43.55±4.26* 32.66±3.94*

DA-5513 62.45±6.55* 49.88±6.99* 36.85±6.25* 34.56±3.84* 29.68±3.74*

Control, ethanol-treated control; UK, treated with ethanol and UK; YM, treated with ethanol and YM; DA-5513, treated with ethanoland DA-5513.The data represent mean±standard deviation (n=8).*P<0.01 vs Control

Figure 1. Effects of DA-5513 on serum (A) AST activity and (B) ALT activity in chronic ethanol-treated rats. CON, control diet; ED,alcohol diet; ED+UK, alcohol diet with UK; ED+DA-5513, alcohol diet with DA-5513. The data represent mean±SD (n=10);**P<0.01 and *P<0.05.

Figure 2. Effects of DA-5513 on serum (A) TG and (B) T-CHO levels in chronic ethanol-treated rats. CON, control diet; ED, alcoholdiet; ED+UK, alcohol diet with UK; ED+DA-5513, alcohol diet with DA-5513. The data represent mean±SD (n=10); **P<0.01 and*P<0.05.



(P<0.01). The administration of UK product after

ethanol diet could reduce the ALT level (P<0.05) but not

AST level in comparison with ED group. In contrast, the

blood samples of the animals treated with DA-5513

revealed significant hepatoprotective activity, as evidenced

by an amelioration of this increase in serum AST and

ALT levels (P<0.01 and P<0.05, respectively).

Effects of DA-5513 on serum and liver lipid profiles

The serum T-CHO and TG levels (Figure 2A and 2B)

increased significantly in the ED group (by approximately

1.2- and 1.8-fold, respectively) (P<0.01). Animals that

received DA-5513 showed a significantly lower level of

both serum TG and T-CHO than that of ED group

(P<0.01 and P<0.05, respectively) while the difference

between UK and ED group was not significant.

In addition, qualitative hepatic TG measurement, which

further confirmed the histological results, demonstrated

that alcohol feeding with Lieber-DeCarly diet greatly

increased the hepatic TG level in mice by 1.8-fold, in

comparison with control group (Figure 3). This elevation

was significant decreased by concomitant administration

of UK product (P<0.05) or DA-5513 (P<0.01).

Effects of DA-5513 on liver histology

The Oil Red O staining technique was employed to

examine lipid droplet accumulation and histological

changes in the liver. As shown in Figure 4, a massive

accumulation of lipid droplets was found in the ED

group. Normal cells exhibited lobular architectures, but

cells in the ED group exhibited panlobular mixed micro/

macro vesicular steatosis and focal clusters of inflammatory

cells, with associated necrosis. In contrast, these

pathologic changes were markedly attenuated in the DA-

5513 group, which was consistent with results of serum

and hepatic TG measurement.

Figure 3. Effects of DA-5513 on hepatic TG in chronic ethanol-treated rats. CON, control diet; ED, alcohol diet; ED+UK;alcohol diet with UK; ED+DA-5513; alcohol diet with DA-5513.The data represent mean±SD (n=10); **P<0.01 and *P<0.05.

Figure 4. Effects of DA-5513 on hepatic lipid levels in chronic ethanol-treated rats. Liver sections were stained with Oil Red O forhistopathological examination. CON, control diet; ED, alcohol diet; ED+UK, alcohol diet with UK; ED+DA-5513, alcohol diet withDA-5513.



Discussion

Heavy alcohol drinking can result in several alcohol-

induced hangover symptoms, which are attributed to the

physiological effects of alcohol and its metabolites. It is

well established that accumulation of acetaldehyde, an

intermediate alcohol metabolite, plays a pivotal role in

the development of hangover [2,3]. In order to determine

the effects of DA-5513 on hangovers, we measured rat

blood alcohol and acetaldehyde levels at different time-

points after the administration of alcohol. Two commercially

available products, YM and UK, were used as positive

control treatments. The administration of DA-5513 and

YM was associated with lower blood alcohol and

acetaldehyde levels over the time-course of the experiment.

However, the reduction in the acetaldehyde level

observed in rats treated with DA-5513 was 20% and

27% greater than that observed in rats treated with YM

and UK, respectively. These findings indicated that DA-

5512 produced more effective reduction of the acetaldehyde

level than YM, demonstrating that it had the potential to

act as a hangover relief beverage.

The liver is the largest internal organ in the human

body and it has many different roles. One of its most

important functions is to filter harmful substances from

the blood. The liver commonly repairs itself by rebuilding

new liver cells when the old ones are damaged. However,

chronic alcohol ingestion can lead to the development of

liver diseases such as fatty liver, alcoholic hepatitis, and

cirrhosis [4]. This liver damage occurs through several

interrelated pathways. The oxidative reactions involved

in alcohol metabolism generate hydrogen, which

converts NAD to NADH, increasing the redox potential

(NADH/NAD) of the liver [28]. This increase in redox

potential inhibits fatty acid oxidation and gluconeogenesis,

promoting fat accumulation in the liver. In addition,

chronic alcoholism induces the microsomal ethanol-

oxidizing system to break down alcohol, mainly in the

endoplasmic reticulum [29]. This pathway, where

cytochrome P450 2E1 is the main enzyme, can account

for 20% of alcohol metabolism. This enzyme is

upregulated by chronic alcohol use, and generates free

radicals and harmful reactive oxygen species via the

oxidation of nicotinamide adenine dinucleotide phosphate

(NADPH) to NADP [28]. This oxidative stress promotes

hepatocyte necrosis and apoptosis, and lipid peroxidation,

which causes inflammation and fibrosis. Inflammation is

also exacerbated by acetaldehyde, which can bind

covalently to cellular proteins, forming antigenic adducts

[30,31].

Apart from the inconvenient symptoms of hangover,

long-term consumption of alcohol in large quantities is

the leading cause of liver disease and hepatic steatosis.

Alcoholic fatty liver disease results from the deposition

of fat and the accumulation of TG in liver cells. The

potential pathophysiologic mechanisms involved in fatty

liver include a reduction in mitochondrial fatty acid β-

oxidation, increased endogenous fatty acid synthesis or

enhanced delivery of fatty acids to the liver, and deficient

incorporation or export of TG as very low-density

lipoproteins [32,33]. The Lieber-DeCarli liquid diet

model is used to induce alcoholic fatty liver disease in

animals, where it causes liver injury, steatosis, and

oxidative stress. This model has been used to investigate

the relationship between alcohol and therapeutic agents

[22,34]. Previous data have indicated that feeding mice

with a standard Lieber-DeCarli formula for 2 weeks was

sufficient to induce significant steatosis [22]. The degree

of steatosis, determined by the hepatic TG concentration,

revealed that the liver samples met the criteria for a

clinical diagnosis of steatosis [35]. In our study, mice

treated with the Lieber-DeCarli diet for 4 weeks showed

steatosis, as confirmed by a significant increase in serum

ALT and AST activities, T-CHO, and TG levels. Our

data showed that dietary DA-5513 markedly attenuated

the hepatic steatosis observed in this model, as indicated

by Oil Red O staining, hepatic TG quantification, and

serum measures of AST, ALT, T-CHO, and TG.

Mono-and poly-herbal preparations have been used in

traditional medical systems for the treatment of liver

disease since long before recorded history; some of these

products appear to have positive effects on this

potentially reversible disease. Both basic and clinical

studies have suggested that herbal medicines and their

constituents such as Gynostemma pentaphyllum (Thunb.)

Makino, Panax notoginseng (Burkill) F.H.Chen (saponins),

Crataegus pinnatifida Bunge (penta-oligogalacturonide),

Dioscorea opposita Thunb. (dioscin), Punica granatum

L. (gallic acid), glycyrrhizin, silymarin, Prunus armeniaca

L (kernels), and baicalin may have modest benefits in the

treatment of fatty liver disease [36]. Therefore, DA-

5513, which is composed of several herbal extracts, was

investigated as a hangover remedy that may reduce the

blood alcohol concentration, as well as preventing

alcoholic fatty liver. GMT-ALC-5L (fermented rice

embryo and bean extract) promotes the health of both



habitual alcohol drinkers and non-habitual alcohol

drinkers. This preparation has been demonstrated to

reduce blood alcohol levels and promote liver function

recovery by modulating alcohol-metabolizing enzymes

[37,38]. Turmeric (Curcuma longa) has been used in

traditional medicines as a household remedy for various

diseases, including biliary disorders, anorexia, cough,

diabetic wounds, rheumatism, sinusitis, and hepatic

disorders. In vitro and in vivo animal studies have

provided evidence for the hepatoprotective effects of

turmeric against a variety of hepatotoxic substances,

including carbon tetrachloride, galactosamine, pentobarbitol,

1-chloro-2,4-dinitrobenzene, 4-hydroxynonenal, and

acetaminophen (paracetamol) [39]. These hepatoprotective

effects may stem from the potent antioxidant effects of

turmeric. Dietary supplementation of turmeric in rats

(1% turmeric by weight for 10 weeks) was found to

significantly protect against iron-induced lipid peroxide

formation [40]. Curcumin is also helpful in the relief of

hangover. It exhibited an inhibitory effect on alcohol

intoxication in humans, as evidenced by a reduced blood

acetaldehyde concentration and reduced discomfort [41,

42]. Curcumin significantly reduces plasma low-density

and very low-density lipoprotein levels, reduces T-CHO

levels in the liver, and increases the α-tocopherol level

in rat plasma, suggesting an in vivo interaction between

curcumin and α-tocopherol that may increase the

bioavailability of vitamin E and decrease T-CHO levels

[43]. Silybum marianum (milk thistle) has been used for

centuries as an herbal medicine for the treatment of liver

disease. Silymarin’s hepatoprotective effects involve

several mechanisms including an antioxidant effect,

inhibition of lipid peroxidation, enhancement of liver

detoxification via inhibition of phase I detoxification,

and an increased hepatocyte protein synthesis, thereby

promoting hepatic tissue regeneration [16]. Animal

studies have also demonstrated that silybin could reduce

the conversion of hepatic stellate cells into myofibroblasts,

thus slowing or even reversing fibrosis [16]. Clinical

studies of patients with chronic alcoholic liver disease in

Austria and Hungary demonstrated that silymarin

administration resulted in a normalization of serum liver

enzyme and total bilirubin levels in patients with

alcoholic liver disease, in addition to improved liver

tissue histology [44]. In patients with cirrhosis, long-

term (41 months) administration of silymarin at 420 mg

per day resulted in a significant increase in survival, as

compared to the placebo group [45]. Although most

studies of water chestnuts (Trapa japonica Flerov.) have

focused on their nutritional and ecological value, they

have also been reported to have antioxidant, anti-cancer,

and anti-diabetic effects; these were associated with a

reduction in blood glucose level, and inhibition of α-

amylase and α-glucosidase [15]. Trapa japonica is also

used as an ethno-medicine for the treatment of gastric

ulcer, diarrhea, alcohol hangover, and dysentery [46].

Trapa japonica Flerov. was reported to significantly

inhibit the production of reactive oxygen species, thus

protecting the liver from tert-butyl hydroperoxide (t-

BHP)-induced damage by stabilizing antioxidant systems

and regulating the mitochondrial membranes within liver

cells [47]. In vivo models have indicated that Trapa

japonica Flerov. significantly attenuated t-BHP-induced

increases in serum glutamate oxaloacetate transaminase

and glutamate pyruvate transaminase levels, and in hepatic

malondialdehyde levels [48]. Guarana has also been

used as a tonic for hangovers, neuralgia and menstrual

headaches, leucorrhea, diarrhea, and fevers; this compound

has been shown to prevent DNA damage in carbon

tetrachloride-treated rats [18,20]. Kudzu (Pueraria

thunbergiana) is employed in traditional Chinese medicine

and its major isoflavone constituent, puerarin, has

antioxidant activity and a variety of biological actions in

cardiovascular disease, gynecological disease, osteoporosis,

cognition, and diabetic nephropathy [49]. Studies of

Pueraria flos showed that it increased the acetaldehyde

removal rate in both rats and humans after alcohol

consumption, and reduced hangover symptoms [50].

The kudzu vine is potentially highly beneficial in the

treatment of liver damage, as it scavenges reactive free

radicals and boosts the endogenous antioxidant system.

Kudzu vine extract significantly reduced the cytotoxicity

and production of reactive oxygen species induced by t-

BHP in vitro and lowered the plasma levels of ALT and

AST in a rat model of carbon tetrachloride-induced

hepatotoxicity [15]. Another ingredient that may help to

counteract the effects of heavy alcohol drinking is honey.

Honey contains fructose, a sugar that promotes alcohol

metabolism [51]. Furthermore, honey has considerable

anti-inflammatory, antioxidant, and antitumor activities,

and plays a key role in normalizing kidney function and

protecting the liver from a range of toxic agents [52].

Consistent with these findings, the combination of these

herbal ingredients in DA-5513 significantly ameliorated

hepatic steatosis, as evidenced by its effects on hepatic

TG, serum ALT and AST activities, and serum T-CHO



and TG levels.

In summary, these findings indicated that DA-5513

produced beneficial effects on alcohol metabolite levels

and alcoholic fatty liver in rats. Further studies are

required to investigate the antioxidant activity of this

preparation, and its effects on lipid mechanism, in rats

administered alcohol.

Acknowledgments

This research was supported by the Yeungnam

University research grants in 2017.

Conflict of interests There is a conflict of interest

regarding the publication of this manuscript. JY Yu, HG

Park, and JH Jun are employees of the Dong-A

Pharmaceutical Co. Ltd. The other authors have no

conflicts of interest to declare.

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