Nghiên cứu một số đặc điểm lâm sàng, cận lâm sàng và nồng độ một số cytokin huyết tương trên bệnh nhân mắc bệnh gan mạn do rượu tt tiếng anh

THAI NGUYEN UNIVERSITY
UNIVERSITY OF MEDICINE & PHARMACY

LE QUOC TUAN

STUDY OF CLINICAL AND SUBCLINICAL
CHARACTERISTICS AND CYTOKINE LEVELS IN
PATIENTS WITH ALCOHOLIC CHRONIC LIVER
DISEASE

Major: Internal Medicine
ID: 97.20.107

DISSERTATION ABSTRACT


THAI NGUYEN - 2018


Dissertation was completed at: Thai Nguyen University of
Medicine & Pharmacy
Advisors:

1. Assoc.PhD. Tran Viet Tu
2. Assoc.PhD. Nguyen Ba Vuong

Reviewer 1: Assoc.PhD.
Reviewer 2: Assoc.PhD.
Reviewer 3: Assoc.PhD.

Dissertation will be defensed in front of dissertation’s university

council at: h:

201.

Held in: Thai Nguyen University of Medicine & Pharmacy

The information from this thesis can be found at:
- Vietnam National Library
- Library of Thai Nguyen University
- Library of Thai Nguyen University of Medicine & Pharmacy


1

INTRODUCTION
Alcohol Liver Disease (ALD) is the result of harmful alcohol
abuse and prolonged exposure. The first stage of ALD is
asymptomatic, reversible if alcohol withdrawal occurs, but the later
stages of ALD can not be reversed, usually resulting in death due to
esophageal varices. There is no radical treatment except liver
transplantation. It affect the quality of life of patients, but also have a
great impact on the socio-economic development.
Studies show that altered immunity and inflammation are key
factors contributing to the progression of ALD. Mediators of the
immune system, such as cytokines or inflammatory factors, are
primarily involved in the stages of the disease. In ALD, the use of
chronic alcohol activates Kupffer cells through receptors, increases
interleukin-1 (IL-1) production, and alpha tumor necrosis factor
(TNF-α), contributes to confusion, hepatic dysfunction, necrotizing
fasciitis, hepatic cellular dysfunction, progressive liver fibrosis, and

cirrhosis. Studies of alcoholic hepatitis patients showed TNF-α, IL-1,
IL-8, leukemia, mononuclear activation and macrophage. This further
demonstrates that inflammation is a key factor in the progression of
ALD.
Inflammation is a major factor contributing to the development
of ALD, and therapies that inflammation are a reasonable strategy.
Understanding the role of some cytokines in ALD stages helps to
detect new therapies that inhibit inflammation at an early stage and
fibrosis in the later stages of the disease is actually beneficial to slow
down progression of the disease.


2

In Vietnam, the increasing number of patients with alcoholic
liver disease is a matter of concern and concern due to the increasing
use of alcohol in Vietnam. However, studies on the association of
cytokines with clinical, subclinical and clinical characteristics in
patients with alcoholic liver disease have not been studied by
researchers. For that reasons, we study: "Study of some plasma
cytokines in patients with alcoholic liver disease" with the aim:
1. Describe the clinical, subclinical characteristics and TNF-α,
IL-12, IL-1β, TGF-β in plasma in patients with chronic liver disease
caused by alcohol
2. Analysis of the relationship between TNF-α, IL-12, IL-1β,
TGF-β in plasma with clinical, subclinical and clinical features in
patients with chronic liver disease caused by alcohol.
SCIENTIFIC SIGNIFICANCE
Emphasizing the role of some cytokines in the diagnosis,
prognosis, providing useful evidence is the scientific basis for the

application of cytokine therapy in the treatment of alcoholic liver
disease.
PRACTICAL SIGNIFICANCE
Determination of TNF-α, IL-12, IL-1β, TGF-β levels in plasma
in patients with alcoholic hepatitis and alcoholic cirrhosis.
Evaluate the relationship between these 4 cytokines and some
clinical and laboratory characteristics in patients with ALD.
SUMMARY OF NEW CONTRIBUTIONS OF DISSERTATION
The first thesis studied 4 cytokines in patients with ALD
plasma.


3

Contributing additional information about the cytokine role
in alcoholic liver disease is the scientific basis for the use of
cytokine-based therapies for the treatment of alcoholic liver disease.
Suggestting use of biomarkers for clinical use to detect
inflammation and fibrosis of the liver.
The layout of the dissertation:
+ The dissertation consists of 102 pages including of the following
sections: the introduction (2 pages): chaper 1: overview (37 pages):
chapter 2: objects and methods (14 pages): chapter 3: results (17
pages): chapter 4: discussion (22 pages): conclusions (2 pages):
petition (1 pages). The dissertation consists of 52 tables, 12 pictures.
The theis references include 102 references consist of 7 Vietnamese
document, 95 English document.
+ The three related articles to the dissertation have been published in
the Vietnam Medical Journal.

4

Chapter 1. OVERVIEW
1.1. Alcoholic liver disease
Epidemiology: Alcohol abuse is prevalent throughout the world,
with an estimated 18 percent of adults in the United States. In 2010,
alcoholic cirrhosis caused 493,300 deaths (accounting for 1% of total
deaths). In the United States, the National Institute of Health
estimates that in 2009, there were more than 31,000 deaths from
cirrhosis and in which cirrhosis accounted for 48% of deaths. The
incidence of alcoholic liver disease is higher in areas with high levels
of alcohol consumption per capita. Areas with high rates of alcohol
consumption and alcoholic liver disease include Eastern Europe,
Southern Europe, and the United Kingdom. Countries with large
numbers of Muslims have the lowest rates of alcohol consumption
and alcoholic liver disease. The United States has an average
consumption of 9.4 L/adult/year, compared with 13.4 L/year in
England and 0.6 L/year in Indonesia.
1.1.2. Clinical features, laboratory results of alcohol liver
disease
In most cases, the clinical is less or asymptomatic in the early
stages and in the compensated period. Clinical manifestations of
ALD vary from asymptomatic or mild to fatal cirrhosis. Therefore,
the diagnosis is highly dependent on clinical signs, different tests and
invasive or non-invasive techniques.
Typical ALD:
- Anorexia, nausea, vomiting, discomfort, weight loss, abdominal
pain and jaundice.
Fever sometimes up to 390C, 50% of cases.

5

- Examination: most have large liver, pain, one third of cases
have large spleen.
- More likely: ascites, edema, hemorrhage, hepatic brain
syndrome.
Symptoms of jaundice, ascites, and hepatic brain syndrome can
be reduced by abstaining from alcohol. If the patient continues to
drink alcohol and poor nutrition can lead to repeated episodes with
manifestations of decompensated cirrhosis, resulting in death.
Some patients with early signs of alcohol abuse such as salivary
gland dysfunction, body weakness, malnutrition, may have peripheral
neurological signs, but usually patients without symptoms and
Reluctance to admit alcohol may be the cause of abnormal liver
function.
During clinical examination of patients with cirrhosis, typical
skin manifestations of liver disease include: cardiovascular disease,
palmar edema and glossy tongue. Jaundice, hepatic brachial
syndrome, ascites and edema may also be seen in patients with endstage liver disease.
Consider ALD when patients have a history of excessive
drinking ( > 40-50 g/day) and clinical abnormalities and abnormal
test indices suggest liver damage. However, when drinking history is
often forgotten, it is often necessary to use indirect alcohol screening
tools.
In patients with alcoholic liver disease, alcohol withdrawal
symptoms, delirium, hypertension, mild fever, abdominal pain,
paranoid hallucinations and hallucinations, have been reported after
72-96 hours of withdrawal. .

6

AST/ALT > 2. Increasing AST < 500 U/L, increasing ALT < 300
U/L. Increasing GGT . MCV > 100 fL.
Histopathology:

Mallory

hepatic

degeneration,

Mallory

syndrome, giant mitochondria, inflammatory cell infiltration, liver
fibrosis and fatty liver.
1.4. Definitive diagnosis of alcoholic liver disease
According to the guidelines of the American Liver Disease
Research Association (AASLD - 2010): Diagnosis based on a history
of alcohol use (screening alcohol use of the World Health
Organization, AUDIT - WHO), clinical symptoms of liver disease,
and abnormal liver enzymes. Liver biopsy helps diagnose the cause,
and identifies the stages of liver damage.
Observational studies showed that serum TNF-α levels as well as
those of the liver increased in patients with alcoholic hepatitis, and
were correlated with the severity of the disease. Apply this scientific
basis to use TNF-α inhibitor in the treatment of patients with ALD.
1.5. The role of some cytokines in alcoholic liver disease

Serum IL-12 levels are increased in patients with alcohol
poisoning, alcoholic hepatitis, alcoholic cirrhosis. IL-12 is highest in
patients with alcoholic hepatitis and gradually decreases with alcohol
abstinence.
TGF-β is central in chronic liver disease, which is related to the
progression of the disease, from initial liver damage through
inflammatory and fibrosis reactions leading to cirrhosis and
hepatocellular carcinoma. TGF-β activates the production of collagen
from astrocytes. Damage to the liver causes TGF-β to enhance
astrocytomatic regulation and activate fibroblasts leading to a wound
healing

response,

including

myofibroblast

and

extracellular


7

deposition. Recognizing as a major profibrogenic cytokine, TGF-β
signaling pathways are associated with the inhibition of progression
of liver disease.
Numerous data indicate that the important role of IL-1β in
alcoholic liver damage depends on the formation and activation of

the inflammasome. It relates to the progression of the disease.
Chapter 2. MATERIAL AND METHODS
2.1. Focus group
2.1.1. The study group
- 95 inpatients with ALD who were treated at the Gastrointestinal
Department of Thai Nguyen Nation Hospital and 103 Military
Hospital; 40 healthy students of Military Medical University.
- Time: From January 2015 to December 2015.
Inclusion criteria
* Study group:
95 inpatients had been diagnosed with ALD according to the
guideline of the American Association for the Study of Liver
Diseases (AASLD) 2010: The diagnosis of ALD is based on a
combination of features, including a history of significant alcohol
intake, clinical evidence of liver disease, and supporting laboratory
abnormalities.
- The histological features of alcohol-induced hepatic injury vary,
depending on the extent and stage of injury. These may include
steatosis (fatty change), lobular in-flammation, periportal fibrosis,
Mallory bodies, nuclear vacuolation, bile ductal proliferation, and
fibrosis or cirrhosis.
- Patients who agreed to take part in the research.


8

* Control Group:
40 healthy students of Military Medical University were
interviewed to investigate alcohol abuse based on the AUDIT
questionnaire; comprehensive clinical examination and clinical

examination; Make laboratory tests to detect and eliminate liver
disease. If they are really healthy (without Hepatitis B and Hepatitis
C, perfectly healthy, liver function tests are normal) agree to
participate in research. They will be selected for the study.
2.1.2. Criteria for exclusion of disease group
The patients were excluded from the study as follows: Simple
steatosis and non alcoholic liver disease of any etiology (drug use,
bile clogging, cancer, hepatitis B, hepatitis C), liver disease patients
with unknown etiology, contraindications for liver biopsy. Do not
agree to participate in research.
- Patients are infected, fungal, virus, allergic, autoimmune.
2.2. Method
2.2.1.Design research: prospective, descriptive, cross-sectional,
comparative itself control.
2.2.2. Template size and selection:
- Sample size were calculated by the equation:

In which:
n is the minimum number of participants diagnosed with ALD
Z is confidence level, 95% level of confidence used, Z= 1.96
: Mean cytokine index in patient with ALD taken from a previous
study.


9
:Standard deviation of mean cytokine index taken from a previous
study.
: Relative precision, selection of  = 0.1.
According to González-Reimers et al, TNF-α in the ALD group was
7.18 ± 3.51 pg/ml, appling this equation. The minimum sample size in

this study was: n = 57, we estimated that theoretical sample size is 95
patients.
2.2.3. Research steps
2.3.3.1. Select the patient
All patients who are diagnosed with ALD in Gastrointestinal
Department were eligible for case selection and were not included in
the exclusion criteria.
2.3.3.2. Clinical examination
- All objects will be examined and recorded in medical records which
have been designed specifically to gather and identify the following
information:
- Screening for alcohol use: AUDIT questionnaire.
2.3.3.5. The technique of testing TNF-α, IL-12, TGF-β, IL-1β in blood
- TNF-α, IL-12, TGF-β, IL-1β in blood was measured by using
ELISA kit supplied by Wkea-China. The plasma samples determine
the cytokines index were stored at -800C. Use ELISA reader–
Diagnostic Automation, USA at Military Medical University.
2.3.3.6.Liver biopsy
Liver biopsies were performed with ultrasound guidance. It’s
Germany's Pajunk automatic biopsy gun in the study. The liver tissue
sample was evaluated by doctors of the Department of Anapath Military Hospital 103.
2.3. Study index


10

- Identify the association between 4 cytokines and some clinical
features and laboratory results.
- Association with age, sex, history of alcohol abuse and
alcoholism, some clinical symptoms and laboratory results.

- Association with stage of liver fibrosis by Metavir classification.
- Association with the characteristic of ALD histological lesion in
results.
2.4. Evaluation criteria used in the study
- AUDIT questionnaire - WHO was used to detect alcohol dependence
or abuse: To score the AUDIT questionnaire, sum the scores for each of the
10 questions. A total 8 for men up to age 60, or 4 for women, adolescents, or
men over age 60 is considered a positive screening test.
- Stage of liver fibrosis by Metavir classification.
2.5. Data processing
Data was coded and analyzed by SPSS software version 20.0
+ Descriptive variables: mean, median, standard deviation, min
and max value.
+ Comparing 2 proportions: χ2 test, with statistical significant
at p < 0.05.
+ Comparing 2 means: Student t-test, with statistical significant
at p < 0.05.
Chapter 3. RESEARCH RESULT
95 patients were researched in the Gastrointestinal Department of
Thai Nguyen Central Hospital and Military Hospital 103 from 1/2015
to 6/2017, the results are as follows


11

3.1. Clinical, subclinical characteristics and TNF-α, IL-12,
IL-1β, TGF-β in plasma in patients with chronic liver disease
caused by alcohol.
Table 3.1. Age characteristics in patients with ALD
Age group

Number

Ratio (%)

< 45
45-59
60-74
≥75
Tổng
average age

30
48
15
2
95

31,6
50,5
15,8
2,1
100
50,65

The age group of 45-59 was the most common, accounting for
50,5%. The average age of the researched patients: 50,65. There were
95 male patients and no female patients.
Table 3.2. Clinical characteristics in patients with ALD
Clinical characteristics Number

Ratio (%)
(n =95)
61
64,2
Anorexia
18
18,9
Hematemesis
60
63,2
Hepatomegaly
Collateral circulation
Jaundice
Vasodilatation

14
63
29

14,7
66,3
30,5

- The number of patients having hepatomegaly accounts for
63,2%. The number of patients having jaundice accounts for 66,3%.


12

Table 3.5. Evaluation of liver enzyme in patients with ALD

Liver enzyme test
Number (n=95)
Ratio (%)
Nomal
2
2,1
increasing
89
93,7
AST (U/L)
ALT < 500
≥ 500
4
4,2
198,19
±
155,05
X ± SD
Nomal
24
25,3
increasing
63
66,3
ALT (U/L)
ALT < 200
≥ 200
8
8,4
85,86

±
71,06
X ± SD
Nomal
2
2,1
GGT (U/L)
increasing
93
97,9
749,10 ± 716,18
X ± SD
The mean AST was 198,19 ± 155,05 U/L. 77,1% of patients had
increased ALT < 200 U/L. The mean ALT was 85,86 ± 71,06 U/L.
The mean GGT was 749,10 ± 716,18 U/L.
Table 3.6. Histological findings in ALD
Histological Findings in ALD
Number
(n=95)
21
Large-droplet fat
Features
of
8
Small-droplet fat
fatty liver
66
Mixed
Steatosis
grade

Location
fatty liver

5-33%
34-66%
> 66%
of Zone 1
Zone 2
Zone 3

Ratio
(%)
22,1
8,4
69,5

25
40
30
91

26,3
42,1
31,6
95,8

68
66

71,6

69,5


13

F0
F1
Fibrosis stage
F2
(n=95)
F3
F4
Significant fibrosis
Fibrosis grade (≥ F2)
(n=95)
Severe fibrosis (≥ F3)
Cirrhosis (F4)
Lipogranuloma
Foamy degeneration
Hemosederosis
Some
indicators
of Mallory-Denk
histopathology Bodies
(n=95)
Megamitochondria
Eosin hepatocellular
change

14

20
25
24
12

14,7
21,1
26,3
25,3
12,6

61

64,2

36
12
9
80
54

37,9
12,6
9,5
84,2
56,8

61

64,2

59

62,1

60

63,2

The grade of steatosis of 34%-66% accounted for 42,1%, the
fatty liver zone 1 accounted for 95,8%. Stage of fibrosis F2
accounted for 26,3%. Foamy degeneration accounted for 84,2%, the
megamitochondria accounted for 64,2%, the Mallory body accounted
for 60.2%. Significant stage of fibrosis (≥ F2) accounted for 64,2%.


14

Table 3.8. Results of TNF-α, IL-12, IL-1β, TGF-β indicators in
patients with ALD and the controls
Cytokine indicators
TNF-α
(pg/mL)

IL-12
(ng/L)

IL-1β

n

95% CI

Median

Study subjects 95

110,61 - 326,42

173,64

Controls

153,71 - 166,71

158,23

24,45 - 32,10

27,47

40

Study subjects 95
Controls

40

Study subjects 95

-

4,00

13,42 - 15,34

14,36
< 0,001

Controls

(ng/L)

< 0,001

< 0,001

(ng/L)

TGF-β

p

40

Study subjects 95

3,19 - 3,30

3,19

1126,43 - 1320,91

1191,46
< 0,001

Controls

40

79563,90- 666819,53

110829,44

There was a statistically significant decrease in the
plasma TGF-β in patients with alcoholic liver disease as
compared to controls.
There was a statistically significant increase in the plasma
TNF-α, IL-12, IL-1β in patients with alcoholic liver disease as
compared to controls.


15

3.2. The relationship between levels of TNF-α, IL-12, IL-1β,
TGF-β in plasma with clinical, subclinical features in patients
with chronic liver disease caused by alcohol.
Table 3.14. Relationship between

TNF-α

in plasma with

histological findings in ALD
Histological findings in ALD

95% CI

Median

F0

14

146,37 - 251,43

164,17

F1

20

145,81 - 190,02

155,97

F2

25

147,50 - 185,40

166,71

F3

24

149,76 - 158,80

153,72

F4

12

154,28 - 312,77

168,12

Disease

Hepatitis

83

151,74 - 166,71

157,10

stage

Cirrhosis

12

152,59 - 362,41

168,12

Fibrosis
stage

TNF-α

n

(pg/mL)

The median TNF-α decreased in the hepatitis group (157,10

pg/mL) was lower than in the cirrhosis group (168,12 pg/mL), the
difference was significant (p < 0,05).

p

> 0,05

< 0,05

16

Table 3.26. Relationship between IL-1β in plasma with
histological findings in ALD
Histological findings in

n

ALD

Median

F0

14

11,30 - 17,65

15,55

F1

20

10,88 - 19,28

16,11

F2

25

11,42 - 16,60

14,09

F3

24

10,60 - 15,23

13,22

F4

12

11,16 - 38,06

13,81

Disease

Hepatitis

83

13,21 - 15,89

14,36

stage

Cirrhosis

12

11,16 - 27,61

13,81

Fibrosis
stage

IL-1β

95% CI

(ng/L)

p

> 0,05

< 0,05

The median IL-1β in the cirrhosis group (13,81 ng/L) was lower
than in the hepatitis group (14,36 ng/L), the difference was significant

(p < 0,05).
Table 3.31. A Relationship between TGF-β in plasma with
histological findings in ALD
Histological findings in ALD

TGF-β
(ng/L)

Features of fatty
liver

Steatosis
grade

n
Largedroplet
fat
Smalldroplet
fat

95%CI

Median

988,42 - 1542,33

1246,59

1119,54 - 1460,42

1289,92

p

21

8

Mixed

66

1083,58 - 1320,91

1169,89

Mild

25

1119,54 - 1460,42

1246,59

Moderate

40

1050,03 - 1474,32

1221,43

> 0,05

> 0,05


17

Severe

30

1059,61 - 1344,88

1136,33

No

4

-

1089,58

Yes

91

1138,30 - 1320,91

1208,24

No

27

1059,61 - 1459,97

1154,26

Yes

68

1119,54 - 1344,88

1199,85

No

29

1119,54 - 1492,30

1270,57

Yes

66

1083,58 - 1321,09

1169,89

No

86

1119,54 - 1342,48

1181,87

Yes

9

1049,21 - 1474,32

1246,59

Foamy

No

36

1229,59 - 1512,69

1331,70

degeneration

Yes

59

1049,21 - 1191,46

1124,34

No

15

1145,91 - 1491,11

1239,40

Yes

80

1091,11 - 1344,88

1163,27

Mallory-Denk

No

41

1131,53 - 1343,68

1208,24

Bodies

Yes

54

1069,12 - 1370,26

1181,87

No

34

1018,86 - 1460,42

1163,27

Yes

61

1126,43 - 1344,88

1208,24

No

35

1232,21 - 1491,11

1342,48

Yes

60

1041,64
1210,07

1121,94

Zone 1

Zone 2

Zone 3

Lipogranuloma

Hemosederosis

Megamitochondria
Eosin
hepatocellular
change

-

> 0,05

> 0,05

> 0,05

> 0,05

> 0,05

> 0,05

> 0,05

> 0,05

< 0,05

The median TGF-β in the Eosin hepatocellular change group
(1121,94 ng/L) was lower than in the without Eosin hepatocellular change
group (1342,48 ng/L), the difference was significant with p < 0,05.

18

Table 3.32. A Relationship between TGF-β in plasma with
histological findings in ALD
Histological findings in ALD

TGF-β
(ng/L)

Fibrosis
stage

Disease
stage

n

F0

14

95%CI
1145,91 - 1512,69

Median
1084,38

F1

20

1062,01 - 1460,42

1136,33

F2

25

1038,04 - 1581,31

1172,28

F3

24

1016,46 - 1344,88

1289,92

F4

12

997,28 - 1529,47

1290,94

Hepatitis

83

1141,12 - 1325,70

1210,64

Cirrhosis

12

997,28 - 1529,47

1084,38

p

< 0,05

< 0,05

There was a relationship between TGF-β in plasma with
disease stage and fibrosis stage in ALD, the difference was
significant with p < 0,05.
Chapter 4. DISCUSSION
4.1. Clinical, subclinical characteristics and TNF-α, IL-12,
IL-1β, TGF-β in plasma in patients with chronic liver disease
caused by alcohol.
4.1.1. General characteristics
Characteristics of age

Our results showed that the age group of 45-59 were the most
common 50,5%. the mean age of the group was 50,65 years, our results
was also similar to the authors, most alcoholism were at working age, so
alcoholism greatly affected the socio-economic development.
Characteristics of sex


19

There were 95 male patients and no female patients, because of
the collection of data in military hospital. The results of the study
showed that the proportion of women with ALD in Vietnam was
lower than in foreign country.
4.1.2. Clinical features
The most common clinical symptoms were fatigue (84,2%),
jaundice (66,3%), anorexia (64,2%), enlarged liver (63,2%).
Symptoms of fatigue, insomnia, poor appetite, jaundice, abdominal
pain, gastrointestinal disorders are common symptoms with high
rates.
4.1.3. Subclinical characteristics
93,7% of patients had an increasing AST but less than 500 U/L.
The mean AST was 198,19 ± 155,05 U/L. 66,3% of patients had
increasing ALT but less than 200 U/L. The mean ALT was 85,86 ±
71,06 U/L. 97,9% of patients had a increasing GGT. The mean GGT
were 749,10 ± 716,18 U/L.
In this study, most of patients had fatty liver. According to the
documents, about 90% of people drinking alcohol with fatty liver.
- Mixed liver fatty was commonly seen in the study subjects
(69.5%). According to the documents, the type of large droplet fatty
or mixed liver fatty was commonly seen in alcoholic fatty liver.

- Steatosis grade: the results of our study showed that the
moderate fatty liver was higher proportion than the mild and severe
fatty liver. The steatosis zone 1 accounted for 95.8%.
- Our results showed that the foamy degeneration accounted for
84,2%), megamitochondria accounted for 62,1%, Mallory body
accounted for 64,2% were common histological features. According
to the medical documents, Mallory body (found in 76% of ALD


20

patients), foam degeneration, megamitochondria were histologic
features in ALD patients.
Our study showed that liver fibrosis F2 (26.3%) and F3 (25.3%)
accounted for the highest proportion, and fibrosis F4 accounted for
the lowest proportion.
Of the 95 subjects, the majority of the study subjects were
alcoholic hepatitis, accounting for 87.4%. Significant levels of liver
fibrosis accounted for 62.1%.
4.1.4.Characteristics of TNF-α, IL-12, IL 1β, TGF- β
Of the 95 patients studied, plasma TNF-α median was 173,64
pg/mL; plasma IL-12 median was 27,47 ng/L; plasma IL-1β median
was 14,36 ng/L and plasma TGF-β median was 1191,46 ng/L.
Of the 40 controls, plasma TNF-α median was 158,23 pg/mL;
Plasma IL-12 median was 4,0 ng/L; plasma IL-1β median was 3,19
ng/L; plasma TGF-β median was 110829,44 ng/L.
The cytokine levels in our study group was very high and nonstandard delivery, so we used the median for comparison between the
study groups and control groups as well as for the relationship
analysis.
4.2. The relationship between levels of TNF-α, IL-12, IL-1β,

TGF-β in plasma with clinical, subclinical features in patients
with chronic liver disease caused by alcohol.
4.2.1.TNF-α
TNF-α can play an independent role in alcoholic liver disease by
promoting the hepatopancreas process. Studies show that alcohol
increases the sensitivity of the hepatocyte to the cytokine's toxic
effects. Of the cytokines identified in alcoholic liver disease, TNF-α
was most closely related to the severity of the disease. In patients


21

with alcoholic hepatitis treated, improvement in clinical presentation
is associated with decreased cytokine levels in the blood. In addition,
TNF-α inhibitors have also been shown to be effective in the
treatment of alcoholic hepatitis.
The results of our study showed that TNF-α was significantly
higher in the plasma of the control group than in the control group.
The difference was statistically significant with p < 0,001. This result
is consistent with González-Reimers et al., Mortensen, ZuwaaJagieo.
When analyzing the association between TNF-α levels and clinical
and subclinical features, we did not find out much association. TNF-α
levels was related to the disease stage, TNF-α median in the alcoholic
cirrhosis group (168,12 pg/mL) was higher than those in the alcoholic
hepatitis group (157,10 pg/mL), with p < 0,05.
4.2.2. IL-1β
IL-1β plays an important role in inflammation in alcoholic liver
disease, which is a major factor in inflammation, fat degeneration and
hepatocellular injury.
Our results: plasma IL-1β median was higher in the study group

compared with the control group, the difference was statistically
significant with p < 0,001. Similar to Petrasek (2012), Tung et al
(2012), Naoko et al (2013).
When analyzing the relationship between plasma IL-1β
concentrations and glucose. The level of IL-1β in the group of
alcoholic hepatitis (14,36 ng/L) was higher than in the alcoholic
cirrhosis group (13,81 ng/L), the difference was statistically
significant with p < 0,05. This result is consistent with that of Tung et
al.


22

4.2.3.TGF-β
Astrocytes in the liver directly affect the fibrosis process by
increasing scar formation. The most potent form of collagen I
production is TGF-beta, derived from both paracrine and autocrine
sources; TGF-beta also stimulates the production of fibroneectin and
proteoglycans. Other factors that stimulate collagen I through
astrocytes include retinoids, angiotensin II, IL-1β, TNF-α, and
acetaldehyde. However, the strongest is TGF-β.
Our results: TGF-β levels in plasma were significantly lower in
the study group than in the control group with p < 0,001. Our results
were similar to Szczerbińska et al (2015), Neuman.
In our study, TGF-β was correlated with disease stage and
fibrosis stage: Median TGF-β levels in F3-F4 fibrosis group (1289,92
ng/L – 1290,94 ng/L) was significantly higher than that of F0-F1
liver fibrosis (1084,38 – 1136,33 ng/L) and the difference was
statistically significant with p < 0,05. This finding is consistent with
the results of Lavallard et al.

4.2.4.IL-12
Results of our study: plasma IL-12 concentration was higher in
the study group than control group with p < 0,001. Similar to author
Tung et al (2012), Sowa.
In conclusion, Tung's research has demonstrated that levels of
serum IL-12 reflect different stages of alcoholic liver disease and
may indicate persistent alcohol consumption. It has the potential to
become a biomarker of alcoholic liver disease.