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PSERUM C-REACTIVE PROTEIN AND PLASMA CERULOPLASMIN LEVELS IN MEN WITH UNCOMPLICATED ALCOHOL DEPENDENCE SYNDROME

PSERUM C-REACTIVE PROTEIN AND PLASMA CERULOPLASMIN LEVELS IN MEN WITH UNCOMPLICATED ALCOHOL DEPENDENCE SYNDROME

Thattil AM,1Mendonsa RD2, Shivashankara AR3

2A 1Assistant Professor, Dept of Biochemistry, Yenepoya Medical College, Mangalore

ssociate Professor, Dept of Psychiatry, Yenepoya Medical College, Mangalore

3Associate Professor, Dept of Biochemistry, Fr Muller Medical College, Mangalore

 

Abstract

Background: Physicians world over have been in the search for accurate and inexpensive markers of excessive alcohol use which would both screen and prognosticate patients with alcohol use disorders.

Aim: Current study compares serum CRP and ceruloplasmin levels among males with alcohol dependence syndrome without physical complications and age controlled healthy males without alcohol dependence/ harmful use.

Method: Sample consisted of 32 consecutive alcohol dependent males admitted to a de-addiction facility and 32 age matched controls. In addition to socio-demographic and clinical data, serum C-reactive protein and plasma ceruloplasmin levels were measured in both cases and controls.

Result: The levels of CRP among patients with alcohol dependence syndrome were significantly higher (mean of 3.23 mg/ltr, SD – 5.04 ) than controls (mean  of 0.92 mg/ltr, SD – 0.65 ) [p = 0.016]. Ceruloplasmin levels were also found to be significantly decreased among alcoholics ( mean – 17.19  , SD -7.399 ) as compared to the controls ( mean – 31.5   , SD - 5.077)  [p  <0.0005].

Conclusion: This study observed significant increase in  CRP, and decrease in  ceruloplasmin levels in alcohol dependent males.  Assessment of these biochemical parameters could be of prognostic importance and help in long term monitoring and follow up of alcoholics.

Key Words: C - reactive protein, ceruloplasmin, alcoholism, alcohol biomarkers

 

Introduction

Alcohol dependence syndrome is a common and potentially lethal condition.  Physicians world over have been in the search for accurate and inexpensive markers of excessive alcohol use which would both screen and prognosticate patients with alcohol use disorders. There are many clinical laboratory tests that can differentiate reliably alcoholics from non alcoholics that has important implications for their treatment and management.1Several studies have indicated that laboratory tests are a reliable medium to detect the presence and extent of detrimental effects of alcohol on the body.2,3

 

Although liver enzymes, gamma glutamyltransferase (GGT), aspartate aminotransferase (AST) and alanine aminotransferase (ALT), and the mean volume of the red blood cells (mean corpuscular volume; MCV) have been the conventional markers of excessive alcohol consumption that provide important prognostic information,carbohydrate-deficient transferrin (CDT), mitochondrial AST (mAST) and alpha glutathione-s-transferase (ct-GST) are considered modern potential biochemical markers.4

Changes in plasma protein levels and status of acute phase protein pattern among alcoholics are being keenly studied by researchers worldwide. Two major mechanisms involved in pathogenesis of alcohol-related complications are inflammation and oxidative stress.5Hence, it is important to assess the inflammatory and antioxidant status in alcohol dependent individuals who are vulnerable to complications such as liver disease. C-Reactive Protein (CRP) is an acute-phase protein whose plasma concentrations increase in response to inflammation In the recent times several population based studies have clearly shown a relationship between alcohol intake and CRP.6,7 This finding is interesting because C-Reactive protein is also considered a marker of future cardiovascular death.8

Ceruloplasmin belongs to the family of multicopper oxidases. Due to its ability to react with, and scavange oxygen species such as superoxide and hydrogen peroxide, ceruloplasminis also considered as a type of plasma antioxidant.9Several investigators have studied ceruloplasmin in patients with alcoholism. Studies have shown conflicting results about the serum copper and ceruloplasmin levels in alcoholics.10 Most of these studies concluded that copper and ceruloplasmin were not good biomarkers for early detection of the deleterious effects of alcohol intake in young adult population in the age group of 20 – 40 years.11

Current study compares serum CRP and ceruloplasminlevels among males with alcohol dependence syndrome without physical complications and age controlled healthy males without alcohol dependence/ harmful use.

 

Objectives

1. To estimate the serum levels of C-reactive protein and plasma ceruloplasmin in alcohol dependent males with no physical complications in comparison to healthy controls.

2. To correlate these biochemical parameters with amount and duration of alcohol abuse.

Materials and Methods

Sample consisted of thirty two Alcohol dependent males and thirty two controls. The cases were consecutive alcohol dependent male patients admitted to the de-addiction centre a teaching hospital.

Inclusion criteria were: male inpatients in the first week of their de-addiction programme, age between 18 and 65 years, patients with the clinical diagnosis of alcohol dependence syndrome by the treating psychiatrist.Patients in delirium, those with significant medical conditions like diabetes mellitus, hypertension and ischaemic heart disease, tobacco chewers and chronic smokers were excluded.

Socio-demographic and clinical data were obtained by interviewing patients as well as from clinical case files. Alcohol consumption related data was obtained by using AUDIT, a screening questionnaire for detailed alcohol use history.

Apparently healthy, non-alcoholic, age-matched male volunteers were the controls for the study. CAGE questionnaire was administered to each control subject to exclude those with problem drinking.

Serum C-reactive protein and plasma ceruloplasmin levels were measured by turbidimetric method12 and p-phenylenediaminedihydrochloridemethod13respectively, in both cases and controls.

Statistical Analysis

 All statistical analyses were conducted by using Statistical Package for the Social Sciences

(SPSS V.17.0)

Primary analyses tested for significant differences in biochemical parameters between cases and controls. Following this, T test and Pearson’s correlation were done to look for any significant associations and correlations between socio-demographic variables, alcohol consumption related variables, and the two biochemical parameters.

Results

Socio-demographic/ Clinical Characteristics of the Sample:

The sample size was 64, out of which 32 were patients with Alcohol Dependence Syndrome (cases) and 32 were healthy male volunteers (controls).

1.Age

Age of cases ranged from 23 to 72 years. Mean was 41.72 (SD 11.419).

Age of the controls ranged from 22 to 60 years, with a mean of 42.62 years (SD 10.87).

2.Early Vs Late Onset Alcoholism

Seventeen of the alcoholics had early onset alcoholism (onset of dependence before 25 years), while the remaining 15 had shown dependence on alcohol after 25 years.

3.Duration of Alcohol Use

Duration of alcohol use ranged from 2 to 50 years. Mean duration of alcohol use was 17.50 years with SD of 11.44.

4. Daily Intake of Alcohol

The average daily intake of alcohol among cases was 3.34 quarters of hard liquor (1 quarter=180 ml). It ranged from one quarter/day in some to 6 quarters/day in some of the subjects.

5. Type of Alcohol Used

Twenty four patients (75%) were consuming whisky while the remaining 8 patients were using brandy.

6. Family History of Alcoholism in First Degree Relatives

Thirteen of the patients (40.6%) had at least one first degree relative with alcoholism while remaining 19 (59.4%) of them did not have alcoholism history among first degree relatives.

 

Biochemical parameters

1. Serum CRP

The levels of CRP among patients with alcohol dependence syndrome were significantly higher (mean of 3.23 mg/ltr, SD – 5.04) than controls (mean of 0.92 mg/ltr, SD – 0.65 ). This difference was statistically significant (p = 0.016).

2. Plasma Ceruloplasmin

Ceruloplasmin levels were also found to be significantly decreased among alcoholics ( mean – 17.19  , SD -7.399 ) as compared to the controls ( mean – 31.5   , SD - 5.077)  [p  <0.0005]

 

 

Table 1. Serum C-reactive protein and Ceruloplasmin in Alcohol-dependent Males And Controls

                

Controls

(n=32)

Alcohol-Dependent Males(n=32)

% Difference (when compared to controls)

Significance; P Value

CRP (mg/L)

0.92 ± 0.65

3.23 ± 5.04

+ 251

0.016; Significant

Ceruloplasmin (mg/dl)

31.5 ± 5.1

17.2 ± 7.4

- 45

0.005; Highly Significant

 

Correlations:

Age and Biochemical Parameters

Serum CRP levels showed positive correlations with age (Pearson correlation coefficients of 0.291) whereas plasma ceruloplasmin showed negative correlation with age (Pearson correlation coefficients of -0.098).  However they were statistically non-significant.

Duration of Alcohol Use and Biochemical Parameters

There were positive correlations between duration of alcohol use and serum CRP (R=0.276), whereas plasma ceruloplasmin (R=-0.512) had negative correlations with duration of alcohol use. The negative correlation between plasma ceruloplasmin and duration of alcohol use was statistically significant. (P< 0.01)

Amount of Daily Consumption of Alcohol and Biochemical Parameters

Serum CRP levels had a positive correlation with increasing daily intake of alcohol. Plasma ceruloplasmin negatively correlated with the daily intake of alcohol. However these correlations did not attain statistical significance.

 

Table 2.Correlations between Serum CRP, Plasma Ceruloplasmin, Duration of Alcohol Intake and Amount of daily intake

 

r Value

P Value and Significance

CRP-Ceruloplasmin

-0.105

>0.10; Non-significant

Duration of alcohol use  -CRP

0.276

>0.10; Non-significant

Duration of alcohol use  -Ceruloplasmin

-0.511

<0.05; Significant

Daily intake of alcohol - Ceruloplasmin

-0.26

>0.10; Non-significant

Daily intake of alcohol - CRP

-0.160

>0.10; Non-significant

 

Discussion

Ceruloplasmin, the serum ferroxidase, is an antioxidant and required also for iron metabolism. It is one of the blood plasma factors with antioxidant activity and there is evidence that alcohol–induced oxidative stress and reactive oxygen radicals play a role in cell injury, particularly damage to the liver cells, thus causing chronic alcoholic hepatopathies.15 The present study observed 45% decrease in serum ceruloplasmin in alcohol dependent males when compared to controls. Serum ceruloplasmin levels also showed significant negative correlation with the duration of alcohol use (r=-0.512, p<0.01). This finding supports the results of a similar study by Uhlikovaet al14 who found that apoceruloplamin levels in patients with liver steatosis were moderately decreased in comparison to healthy controls. Uhlikova et al also found that the specific activity of ceruloplasmin in patients was significantly decreased in comparison to controls thus suggesting some problems in copper metabolism in these patients. Our sample of alcoholics consisted of patients who had an average daily consumption of around of 540 ml of hard liquor and an average duration of 17.5 years. Although most of them were heavy drinkers, all our subjects were apparently healthy. Finding of low plasma ceruloplasmin levels in these patients is interesting. It implies that plasma ceruloplasmin might be a sensitive marker of alcohol–induced oxidative stress and cell injury, and its levels significantly decrease much before the clinical features of alcoholic liver disease become apparent.

C-reactive protein (CRP) ware acute phase reactant and used clinically as a marker of inflammatory conditions.  In the present study serum CRP was found to be higher in alcohol dependent subjects, and there was positive though statistically non significant correlation of serum CRP with duration of alcohol use and daily intake of alcohol. In a earlier study, a U-shaped association between alcohol consumption and C-reactive protein (CRP), with the association being stronger among men compared to women.16Several studies have shown inflammation markers such as C reactive protein (CRP) predict future CHD complications even better than all other biomarkers, including low density lipoprotein (LDL) cholesterol.17,18 Although the serum CRP levels among the alcoholics in our study were in the normal range (<10mg/l), a level above 2.4 mg/L has been associated with a doubled risk of a coronary event compared to levels below 1 mg/L.19  Studies have reported that alcoholism induces synthesis of acute phase proteins in liver. Lippi et al. observed increased serum levels of alpha-1 acid glycoprotein, fibrinogen, haptoglobin, and reported that these increases were independent from the existence of liver damage.20 This is very relevant to our findings as the patients in our study were apparently healthy without any physical complications related alcoholism.

Limitations the sample size was smaller.  The cross-sectional design of the study limits our ability to elucidate any cause-effect relationship between the parameters studied. Similar studies employing large sample size are needed to confirm these findings. Longitudinal studies with long-term follow up could help in understanding relationships between alcoholism related clinical factors, CRP and ceruloplasmin.

Conclusion

Chronic alcohol abuse caused significant decrease in plasma ceruloplasmin levels and increase serum CRP levels.  Altered ceruloplasmin level suggests a deranged copper metabolism and tilting of oxidant-antioxidant balance in favour of oxidants. Alcoholism induces synthesis of acute phase proteins in liver including CRP increasing the risk of an adverse coronary event. Assessment of these biochemical parameters could be of prognostic importance and help in long term monitoring and follow up of alcoholics.

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