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ORIGINAL ARTICLE
Year : 2019  |  Volume : 32  |  Issue : 1  |  Page : 231-237

Study of serum and seminal carnitine levels in Egyptian men with chronic hepatitis C


1 Department of Dermatology, Andrology and STDs, Faculty of Medicine, Menoufia University, Shebin El-Khoum, Menoufia, Egypt
2 Department of Medical Biochemistry, Faculty of Medicine, Menoufia University, Shebin El-Khoum, Menoufia, Egypt
3 Department of Dermatology, Andrology and STDs, Ministry of Health, Birket El Sab, Menoufia, Egypt

Date of Submission25-Apr-2017
Date of Acceptance18-Jun-2017
Date of Web Publication17-Apr-2019

Correspondence Address:
Wesam A El-Degwy
Department of Dermatology, Andrology and STDs, Ministry of Health, Birket El Sab, Menoufia
Egypt
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/mmj.mmj_304_17

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  Abstract 


Objective
The aim of the study was to evaluate the serum and seminal carnitine levels in Egyptian men with chronic hepatitis C virus (HCV) infection.
Background
In 2015, HCV still affects a substantial proportion of the Egyptian population, in the 1–59 year age, 5.3 million persons are positive for HCV antibodies. Numerous studies have investigated the benefits of carnitine supplementation on improving the complications of viral hepatitis, like steatosis and hepatic encephalopathy.
Patients and methods
A cross-sectional study was conducted on 35 patients with HCV attending the hepatitis outpatient clinic, in Shebin El-Kom Teaching Hospital, Egypt. Moreover, 20 healthy individuals were also included as a control group. They were all subjected to history taking and serum and semen carnitine level examination.
Results
Results revealed an insignificant difference between patients with HCV and healthy individuals concerning age, residence, and serum urea level. However, a significant difference was found between the groups regarding serum glutamic pyruvic transaminase, serum glutamic oxaloacetic transaminase, creatinine level, prothrombin activity (%), and serum and semen carnitine levels. Moreover, a nonsignificant positive correlation (r = 0.331, P = 0.052) was founded between serum and semen carnitine levels in patients with HCV; in addition, a nonsignificant negative correlation (r = −0.364, P = 0.301) was found between serum and semen carnitine levels in the control group.
Conclusion
Results concluded that patients with HCV had significantly higher serum glutamic pyruvic transaminase, serum glutamic oxaloacetic transaminase, urea, and creatinine level. However, they had significantly lower prothrombin activity and serum and semen carnitine levels compared with the healthy group. Moreover, an insignificant correlation was detected between serum and semen carnitine levels for patients with HCV. So, many studies on various world population with this trend are warranted.

Keywords: carnitine, chronic hepatitis C, semen


How to cite this article:
Attia AM, Yasien HA, Habib MS, El-Degwy WA. Study of serum and seminal carnitine levels in Egyptian men with chronic hepatitis C. Menoufia Med J 2019;32:231-7

How to cite this URL:
Attia AM, Yasien HA, Habib MS, El-Degwy WA. Study of serum and seminal carnitine levels in Egyptian men with chronic hepatitis C. Menoufia Med J [serial online] 2019 [cited 2019 Aug 25];32:231-7. Available from: http://www.mmj.eg.net/text.asp?2019/32/1/231/256098




  Introduction Top


Hepatitis C virus (HCV) infection is a major public health problem, with an estimated global prevalence of 3%. The highest prevalence of hepatitis C virus (HCV) was in Egypt, with 12% among the general population [1] and 40% in persons older than 40 years living in rural areas. HCV genotype 4 is the predominant genotype being isolated from up to 91% of HCV-infected persons in Egypt [2]. The origin of the HCV epidemic in Egypt has been subjected to intravenous schistosomiasis treatment in rural areas in the 1960s–1970s [3]. Hepatitis is a vital inflammation of the liver, which is caused by a virus or results because of an overactive immune system, drugs, alcoholism, chemicals, and environmental toxins [3].

l-Carnitine is an amino acid found in red meat. Carnitine is synthesized in small quantities by the brain, liver, and kidneys [4]. This amino acid plays a role in the release of energy from fat by moving fats to power plants within cells where fats can be burnt as fuel, which are called mitochondria and are responsible cell survive and function [5]. Carnitine is a water-soluble polarized substance that plays a fundamental role in production and dissemination of cellular energy and is involved in numerous metabolic steps [6]. Carnitine facilitates the oxidation of glucose by deleting acyl groups developed by fatty acid β-oxidation [7].

l-Carnitine in seminal plasma plays an important role in maintaining male fertility. There is a potential relationship between carnitine level and semen quality, as fertile males have higher carnitine levels in semen than infertile ones [8]. In addition, results of other studies also observed a positive relation between the concentration of seminal carnitine level and number of spermatozoa and their motility, and morphology [9].

Several studies have examined the benefits of carnitine supplementation on improving the complications of HCV such as steatosis, fatigue, uremia, and hepatic encephalopathy [10].

Although few studies have evaluated the level of serum carnitine in chronic hepatitis [11], to the best of our knowledge, there is no study regarding the correlation between serum and seminal carnitine level in individuals with chronic HCV in the Egyptian population.

This study is aimed at evaluating the changes in the serum and seminal plasma carnitine levels in patients with HCV.


  Patients and Methods Top


The cross-sectional study was carried out on 55 individuals who were divided into two groups: group I included 35 patients with HCV, whereas group II included 20 age-matched and sex-matched healthy individuals who were clinically free of HCV (control group).

All individuals were selected from the hepatitis outpatient clinic, Shebin El-Khoum Teaching Hospital, Egypt, from the period of January 2015 till December 2015.

Ethical consideration

All individuals signed an informed written consent after explaining the objective of the study to them before the initiation of the study. The consent form was developed according to the standard in Quality Improvement System in Ministry of Health and Population in Egypt, which was introduced in all Dermatology, Andrology and STIs centers and units. Moreover, it was modified according to the recommendations of ethical committee in Faculty of Medicine, Menoufia University.

Inclusion criteria

Patients who were confirmed as having HCV based on the serological markers and liver enzymes analysis were included in the study.

Exclusion criteria

Recent history revealing patients taking supplements containing carnitine during the past 3 months, patient receiving medications that alter carnitine metabolism, those with renal impairment, vegetarians, patients with decompensated cirrhosis, and those with multiple organ dysfunctions were excluded from the study.

Individuals were submitted to the following:

  1. History taking: it was done using a predesigned questionnaire through exclusion or inclusion of items; it included personal data such as name, age, and residence
  2. Investigations:


    1. General investigations:


      1. Kidney functions: blood urea and serum creatinine levels were analyzed using the open system autoanalyzer synchron CX5 (Beckman Coulter, Inc. Atlanta Vision Center Atlanta, Georgia, USA)
      2. Liver functions: it included examination of serum glutamic pyruvic transaminase (SGPT) and serum glutamic oxaloacetic transaminase (SGOT) levels, which were analyzed using the open system autoanalyzer synchron CX5 (Beckman Coulter)


    2. Prothrombin activity (%): prothrombin time ratio was calculated using Coagu Chek System (Roche Diagnostics Corp., Indianapolis, Indiana, USA)
    3. Special investigations: it included serum and semen carnitine levels using the ELISA kit (Life Technologies Corporation, Carlsbad, California, United States).


Human l-carnitine ELISA kit

Principle of the assay

This assay employs the quantitative sandwich enzyme immunoassay technique. Antibody specific for l-carnitine are precoated onto a microplate. Standards and samples are pipetted into the wells, and any l-carnitine if present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for l-carnitine is added to the wells. After washing, avidin-conjugated horseradish peroxidase is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells, and the color that develops is in proportion to the amount of l-carnitine bound in the initial step. The color development is stopped, and the intensity of the color is measured.

Assay procedure summary

  1. Reagents, samples, and standards were prepared as instructed by the manufacturer
  2. 100 μl standard or sample was added to each well and incubated for 2 h at 37°C.
  3. The liquid was removed for each well
  4. 100 μl biotin-antibody (1×) was added to each well and incubated for 1 h at 37°C
  5. It was aspirated and washed three times
  6. 100 μl horseradish peroxidase-avidin (1×) was added to each well and incubated for 1 h at 37°C
  7. 90 μl TMB substrate was added to each well and incubated for 1 h at 37°C
  8. 50 μl stop solution was added to each well and read at 450 nm within 5 min.


Statistical analysis

Results were tabulated and statistically analyzed by using personal computer using Microsoft Excel 2016 and SPSS, version 21 (SPSS Inc., Chicago, Illinois, USA). Descriptive analysis included %, mean, and SD. Analytical analysis includes χ2-test, Student t-test, Mann–Whitney test, and correlation coefficient test (Person's test). A value of P less than 0.05 was considered as statistically significant.


  Results Top


In this study, it was found that the age of the patients with HCV ranged from 27.0 to 48.0 years, with mean of 37.57 years, and for the control group, it ranged from 33.0 to 50.0 years, with a mean of 41.70 years. Regarding residence, the highest percentage was from the rural area, with 62.9% of the patients with HCV and 60% of the control group. Moreover, an insignificant difference was found between patients with HCV and healthy groups concerning age and residence [Table 1].
Table 1: Demographic characteristics of the studied groups regarding residence and age

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Results showed that highly statistically differences were detected between studied patients with HCV and healthy groups regarding SGPT and SGOT (P = 0.001). The highest means of SGPT (48.60) and SGOT (43.40) were recorded in patients with HCV as compared with control group, which recorded the lowest mean values of SGPT and SGOT [Figure 1]. Moreover, results illustrated that serum urea level in patients with HCV ranged from 10.0 t o 38.0 mg/dl, with mean ± SD of 27.20 ± 8.11 mg/dl, and it ranged from 20.0 to 36.0 mg/dl with mean ± SD of 27.10 ± 5.30 mg/dl in control group, and the difference between them did not reach significance level at 0.05 [Figure 2]. Regarding serum creatinine level (mg/dl), results found that significant differences (P = 0.001) were detected between patients with HCV and healthy subject groups [Figure 2]. Serum creatinine level ranged from 0.60 to 1.40 mg/dl, with mean ± SD of 1.07 ± 0.23, in patients with HCV, whereas it ranged from 0.50 to 1.10 mg/dl, with mean ± SD of 0.78 ± 0.21 mg/dl, in the control group [Table 2].
Figure 1: Comparison between the two studied groups regarding liver function (SGPT and SGOT). HCV, hepatitis C virus; SGOT, serum glutamic oxaloacetic transaminase; SGPT, serum glutamic pyruvic transaminase.

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Figure 2: Comparison between the two studied groups regarding serum urea and creatinine level (mg/dl). HCV, hepatitis C virus.

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Table 2: Comparison between the two studied groups regarding liver function and kidney function

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Results revealed that all of patients with HCV and healthy controls had negative results for l-carnitine supplementation in the past 3 months. Insignificant differences were detected between cases and healthy subjects regarding l-carnitine level (P = 1.00), whereas statistically significant differences were detected between patients with HCV and control groups regarding prothrombin activity (%) (P = 0.042). The largest mean percent of prothrombin activity (97.20 ± 2.90) was founded in control group as compared with patients with HCV group, which recorded the lowest mean percent (89.37 ± 7.09) [Table 3].
Table 3: Comparison between patients with hepatitis C virus and controls regarding l-carnitine supplementation in the past 3 months and prothrombin activity (%)

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In this study, it was found that the studied groups statistically differed regarding serum carnitine (P = 0.018) and semen carnitine (P = 0.001) levels. Serum carnitine level ranged from 2.01 to 3.99, with mean value of 3.03 ± 0.68, and ranged from 2.30 to 5.11, with mean of 3.66 ± 0.81, in patients with HCV and healthy groups, respectively [Figure 3]. Also, semen carnitine level ranged from 2.01 to 3.94, with mean value of 2.96 ± 0.59, in patients with HCV group, whereas it ranged from 3.52 to 4.88, with mean value of 4.06 ± 0.48, in the control group [Figure 3] and [Table 4].
Figure 3: Comparison between the two studied groups regarding serum and semen carnitine levels. HCV, hepatitis C virus.

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Table 4: Comparison between the two studied groups regarding serum carnitine and semen carnitine levels

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Moreover, results in this study revealed that a nonsignificant weakly positive correlation (r = 0.331, P = 0.052) was present between serum and semen carnitine levels in studied patients with HCV (n = 35). The covariance between them reached to be 0.131 [Figure 4]. In addition, a nonsignificant weakly negative correlation (r=−0.364, P = 0.301) was present between serum and semen carnitine levels in control group (n = 20) [Figure 5]. The covariance between them reached to be − 0.141 [Table 5].
Figure 4: Correlation coefficient (r) between serum carnitine and semen carnitine levels in the studied patients with hepatitis C virus (HCV) (n = 35).

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Figure 5: Correlation coefficient (r) between serum carnitine and semen carnitine levels in the control group (n = 20).

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Table 5: Correlation coefficient (r) between serum carnitine and semen carnitine levels in the studied patients with hepatitis C virus and controls (n=55)

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  Discussion Top


In the current study, it was found that mean age of patients with HCV was 37.57 years and for control group was 41.70 years. In addition, more than two-thirds (62.9%) of the studied patients resided in rural area. An insignificant difference was observed between patients with HCV and healthy controls concerning age and residence. This comes in agreement with El-Hawary et al. [12] who found that mean age of HCV cases was 41.7%. In their study, 71.6% were males and 28.4% females. Over two-thirds (68.2%) of the individuals were from the urban area. Ibrahim et al. [13] found that nearly 60% of the sample were males with age of 43.5 (range: 18–68 years), and 57.7% of them were from the urban area. Cruciani et al. [14] found that females comprised 18% of the placebo group versus 6% in the l-carnitine group.

In the current study, it was found that highly statistically differences (P = 0.001) were founded between studied patients with HCV and healthy groups regarding SGPT and SGOT. Patients with HCV scored the highest mean of SGPT (48.60) and SGOT (43.40) as compared with control group. This may be because l-carnitine supplementation prevents elevation in liver functions, weakness, and hypotension induced by isotretinoin. This is in agreement with El-Shahat et al. [15] who found that SGOT is statistically higher in male and female patients with both high and moderate HCV titer versus control group. This increase in females with high HCV group compared with other groups. In addition, Tutal et al. [16] found that SGOT was much significantly higher in the HCV-positive group (P = 0.017) with an odds ratio value of 1.1 (95% confidence interval: 1.0–1.3), and this confirms the subtle inflammatory state existing within HCV-positive hemodialysis (HD) patients. Moreover, increased SGOT is a sensitive indicator of liver damage, and these results were found by Meyer and Harvey [17]. SGOT and alkaline phosphatase were statistically higher in HCV-positive patients (P = 0.04) as compared with control group.

In the current study, it was found that mean value of serum urea in patients with HCV group was 27.20, which was more than in control group (27.10), and the difference between them did not reach significance level at 0.05. This comes in agreement with Hosny et al. [18] who found that blood urea was slightly increased in HCV-negative HD patients versus HCV-positive patients, in a nonsignificant way. They added that a positive relation that was founded between P-selectin and each of blood urea. On the contrary, significant differences (P = 0.001) were founded between studied groups concerning serum creatinine level. Patients with HCV recorded the highest mean serum creatinine level (1.07) compared with control group (0.78 mg/dl). This comes in agreement with Russell et al. [19] who found the diagnosis of renal disease relies on changes in the urea-nitrogen, creatinine, urine analysis, and measurement of urine output. Hosny et al. [18] found that serum creatinine level was slightly lower in HCV-negative HD patients versus HCV-positive patients, in a nonsignificant way.

Our results showed that all studied patients with HCV and healthy groups had negative l-carnitine supplementation in the past 3 months, and the differences between them did not reach significance level regarding l-carnitine (P = 1.00). This was consistent with Feng et al. [20] who reported that administration of l-carnitine decreased sucrose-induced hypertriglyceridemia and the increase of free fatty acid in rat plasma.

A statistically significant difference was found between the studied patients with HCV and healthy controls regarding prothrombin activity (%) (P = 0.001). Patients with HCV scored the lowest mean percent (89.37) of prothrombin activity compared with the controls. This comes in consistent with Nkontchou et al. [21] who found that the patients infected with HCV had a low activity of prothrombin versus other genotypes. In addition, Marconcini et al. [22] found that lower activity of prothrombin (72.6 ± 11.5 vs. 82.2 ± 16.0%; P = 0.012) was found in patients with HCV.

In the current study, statistically significant differences were founded between patients with HCV and healthy groups regarding serum carnitine (P = 0.031) and semen carnitine (P = 0.001) levels. The highest mean values of serum and semen carnitine levels were observed in the control group (3.66 and 2.96, respectively). On the contrary, the lowest mean values of serum and semen carnitine levels were recorded in patients with HCV (3.03 and 4.06, respectively). This is consistent with Stradaioli et al. [23] who revealed higher seminal carnitine levels in low-motility group versus high-motility group, whereas Tomamichel and Bandhau [24] showed normal range of 440–990 mmol/l. This may be because of different dietary habits and demographic variation, as ~ 75% of the body stores of l-carnitine are derived from the diet. Moreover, Ahmed et al. [7] suggested that l-carnitine level in seminal plasma plays a vital role in maintaining male fertility.

The current study found a nonsignificant weakly positive relation between serum and semen carnitine levels in studied patients with HCV, as well as a nonsignificant weakly negative correlation in the control group. This is in agreement with Gravitz et al. [25] who concluded that there is a potential correlation between carnitine level and semen quality. Similar correlation was also confirmed by Ke et al. [8] with lowest levels in the oligoasthenopermic group.


  Conclusion Top


Results conclude that patients with HCV had significantly higher values for SGPT, SGOT, serum urea, and serum creatinine levels, but significantly lower values for prothrombin activity (97.20) and serum and semen carnitine levels as compared with healthy controls. Moreover, a nonsignificant correlation was detected between serum and semen carnitine levels for patients with HCV. So, many studies on various world population with this trend are warranted.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
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    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5]



 

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