ORIGINAL RESEARCH ARTICLE http://www.eternalpublication.com INTERNATIONAL JOURNAL OF ANATOMY PHYSIOLOGY AND BIOCHEMISTRY IJAPB: Volume: 4; Issue: 9; September 2017 ISSN(Online):2394-3440 Study on Diagnostic Markers of Liver Disease in Non Alcoholic and Alcoholic Patients Published online on 12 th September 2017 www.eternalpublication.com PADMINI PRAKASH HABBU 1 VASUNDHARA DESHMUKH 2 ABDUL KAYYUM SHAIKH 3 1, 2 Tutor 3 Professor & Head Department of Biochemistry Ashwini Rural Medical College, Hospital & Research Center, Solapur, Maharashtra Corresponding Author: Padmini Prakash Habbu S/3 Sukhaswapna Appartment, Behind Mahila Hospital, Hodgi road, Solapur-413003 Maharashtra (India) +917038699499 phabbu18@gmail.com Received: 5 th August 2017; Accepted: 20 th August 2017 How to cite this article: Habbu PP, Deshmukh V, Shaikh AK. Study on diagnostic markers of in non-alcoholic and alcoholic patients. International Journal of Anatomy Physiology and Biochemistry 2017;4(9):1-5. Abstract: Aims: To evaluate the diagnostic markers of in alcoholic and non alcoholic patients. Methodology: The study group included 100 cases among which 50 alcoholic and 50 with non alcoholic while control group included 50 healthy persons. Serum urea, creatinine, total bilirubin, total protein, A/G ratio, serum enzyme activity like AST, ALT, ALP, gamma GT, AST/ALT ratio estimation was done by using kit on mini VIDAS. Results: In present study, patients with either due to alcohol or without alcohol were compared with a group of normal healthy persons. Mean value of urea, creatinine, total bilirubin, total protein, A/G ratio and enzymatic parameters were determined. The drinker alcoholic patients had significantly high urea, creatinine and total bilirubin and significantly lower total protein and A/G ratio compared to other groups. The serum enzyme activity like AST, ALT, ALP, γgt, and AST/ALT ratio were found to be significantly increased in comparison to healthy controls as well as in drinkers in comparison to non drinkers. The AST/ALT ratio, γgt and total bilirubin in non alcoholic correlated with AST/ALT ratio, γgt and total bilirubin in alcoholic which was negatively correlated with each other. Conclusion: Hyperbilirubinemia, hyperurea, hypercreatinine, hypoalbuminemia are common features of alcoholics as well as non alcoholic. Monitoring γgt, ALP, AST and ALT in combination is a sensitive means of detecting severity of liver damage. Keywords: alcoholic, non alcoholic, AST, ALT, γgt Introduction: Alcohol consumption (leading cause of liver cirrhosis) is the world s third largest risk factor for and disability in middle-income countries. Harmful use of alcohol results in approximately 2.5 million deaths each year, with a net loss of 2.25 million lives. Globally 6.2% of all male deaths are attributable to alcohol, compared to 1.1% of female deaths. 1-3 Alcohol-related data remain scarce in India and so far there have been very few scientific studies. 4 Alcohol ingestion is known to produce a variety of metabolic and pathological alterations in the liver, which are especially due to cell metabolic disturbances associated with ethanol oxidation and oxidative stress. 5,6 Serum assay for biochemical 1
markers of the are an integral part of proper evaluation of liver. In general, serum bilirubin level is measured to confirm the presence and severity of the. Several enzymes are being used to distinguish and assess hepatocellular injury. Of these, γ-glutamyl transferase (γgt), Aspartate aminotransferase (AST) and Alanine aminotransferase (ALT) are commonly used as screening tests. 7,8 γgt, the most sensitive and widely employed marker for alcohol abuse. The marked elevations of these aminotransferases reflect severity of active hepatocellular damage. The ratio of AST to ALT (AST:ALT) help determine whether the liver or another organ has been damaged, or help determine if liver damage is related to alcohol dependence. It has been reported that medically diagnosed alcoholics can be differentiated reliably from non-alcoholics using clinical laboratory tests. Moreover, distinguishing alcoholic from nonalcoholic has important implications for treatment and management. In the present study we have investigated changes in biochemical parameters in patients with alcoholic liver injury (both moderate and heavy drinkers) and nonalcoholic s. These parameters were compared with normal values obtained from normal healthy people. Biochemical parameters included common laboratory tests. Materials & Methods: Study area The present study is an observational study of non alcoholic and alcoholic in rural area of Solapur. The study was conducted in Ashwini Rural Medical College, Hospital and Research Centre, Solapur, Maharashtra. The total 100 patients were taken; 50 patients with alcoholic and 50 with non alcoholic from rural area of Solapur. To compared 50 healthy controls were taken. Study design This is a descriptive, hospital-based study. A] Cases This included all confirmed cases of alcoholic and non alcoholic, who were admitted to clinical care unit. Patient with previous history of were also included. B] Controls Age and sex matched controls were recruited either from volunteers or patient admitted. None of them had any evidence of. C] Exclusion criteria applied The individuals who were diabetic, or had essential hypertension, thyroid, nephritic, and pregnant women were excluded from this study. Consent was obtained from every subject. Data was recorded using standard form on age, sex, gender, blood pressure. Blood samples were collected from all the subjects after taking proper consent. The samples were used to performing biochemical investigations like urea, creatinine, total bilirubin, total protein, A/G ratio, serum enzyme activity like AST, ALT, ALP, γgt, AST/ALT ratio. Serum urea were estimated by urease method, serum creatinine by Jaffes kinetic method, total bilirubin by Jendrassik method, total protein by Biuret method, AST and ALT by UV kinetic method, ALP by kinetic method and gamma GT by Szacz and Rosalki kinetic method. Statistical analysis: The analysis of data was done by using student t test and SPSS-17 software. The difference in mean values of various parameters was calculated and expressed in terms of p value. Ethical considerations: This study was approved by the institution ethics committee. Written consent was obtained from all patients and families after full explanation of and potential hazards and expected benefits of the investigation. Patients were informed about any abnormal results of tests performed and will be instructed and treated. Selection of cases and controls 2
Table 1: Cut off levels for different biochemical parameters used in study Biochemical parameters Cut off level Urea > 40 mg/dl Creatinine > 1.5 mg/dl Total bilirubin > 1 mg/dl Total protein 8 mg/dl A/G ratio >2.5 AST 40 U/L ALT >55U/L ALP >115 U/L γgt >30 U/L AST/ALT ratio >2 Results: Table 2: Urea, Creatinine, Total Bilirubin, Total Protein, and A/G ratio of healthy controls, non alcoholic and alcoholic patients Parameter Control Non alc. Urea 26.82±7.13 43.28±6.91 Creatinine 0.92±0.24 1.28±0.20 T. Bilirubin 0.52±0.24 1.36±0.24 T. Protein 7.02±0.78 5.38±0.48 A/G ratio 1.38±0.34 1.83±0.40 Alc. Liver 54.16±6.15 1.82±0.39 2.19±0.38 4.96±0.48 0.65±0.14 P< 0.01 significant highly significant P >0.05 non significant In present study, we evaluated the diagnostic markers of in alcoholic and non alcoholic patients and compare with a group of normal healthy persons. Mean and standard deviations have been expressed. Serum urea, creatinine, total bilirubin, total protein, A/G ratio and enzymatic parameters were determined. The drinker alcoholic patients had significantly high urea, creatinine and total bilirubin and significantly lower total protein and A/G ratio compared to other groups (Table 2). The serum enzyme activity like AST, ALT, ALP, γgt and AST/ALT ratio were found to be significantly increased in comparison to healthy controls as well as in drinkers in comparison to non drinkers. (Table 3) Table 3: Serum enzyme activity for healthy controls, non alcoholic and alcoholic patients Parameter Control Non alc. liver AST 24.32±5.90 184.38±27.47 ALT 24.7±6.5 89±23.03 ALP 88.08±20.42 119.52±15.48 γgt 15.14±5.78 42.42±6.67 AST/ALP 1.03±0.40 2.14±0.46 ratio Alc. Liver 190.76±37.35 90.24±27.77 154.98±26.84 46.68±7.20 2.13±0.62 P< 0.01 significant highly significant P >0.05 non significant Table 4 shows correlation with AST/ALT, γgt, total bilirubin in alcoholic data with non alcoholic data. AST/ALT ratio was shown to have positive correlation with total bilirubin (r = +0.017) and negative correlation was seen with, γgt (r = -0.036), AST/ALT ratio (r = - 0.109). Positive correlation was perceived between γgt and total bilirubin (r = +0.286), γgt and γgt (r = +0.332), and negateve correlation was seen among t. bilirubin vs t.bilirubin (r = - 0.012). Table 4: Correlation with AST/ALT, γgt, total bilirubin in alcoholic data with non alcoholic data No. Laboratory variable r p value value 1] A) AST/ALT vs AST/ALT -0.109 P 0.001 B) AST/ALT vs γgt -0.036 P 0.001 C) AST/ALT vs T.bilirubin 0.017 P 0.001 2] A) γgt vs γgt 0.332 P 0.001 B) γgt vs T.bilirubin 0.286 P 0.001 3] A) T.bilirubin vs T.bilirubin -0.012 P 0.001 3
Table 5: The De Ritis Ratio and number of patients with alcoholic and non alcoholic No. De Ritis Ratio Alcoholic Non alcoholic 1 AST/ALT < 2 8 21 2 AST/ALT > 2 42 29 Discussion: In the present study, patients with s either due to alcohol or without alcohol were compared with a group of normal healthy persons. The biochemical parameters like urea, creatinine, total bilirubin, total protein, A/G ratio and enzymatic parameters like AST, ALT, γgt, ALP, AST/ALT ratio were determined in this study. Patients with various forms of liver disorders showed hyperbilirubinemia. In the present study, there is an increase of total bilirubin level of serum in both groups of alcoholic as well as non alcoholic patients, respectively. Uptake and excretory functions of liver is constrained depending upon the increasing of bilirubin level of serum. The urea level was found to be increases range in all the tested groups (Table 2). Though the increases in creatinine level of non-alcoholic group was found to be statistically significant in comparison to other groups, but no significant change was observed in alcoholic groups when compared with normal healthy persons (Table 2). The total protein and A/G ratio was found to be significantly decreased in both non alcoholic and alcoholic patients compared to healthy control. Das SK et.al 9 also found decreased protein and A/G ratio. Ethanol consumption slows down the rate of hepatic protein catabolism. The activities of AST and ALT were increased in alcoholic and non alcoholic when compared with healthy controls which is significant (p < 0.01). The ratio of AST/ALT ratio was also elevated in both group as compared to control subjects (p < 0.001)(Table 3). Ratio of AST/ALT >2 implicit the presence of alcoholic live. In our study also out of 50 alcoholic patients, 42 patients and non alcoholic 29 patients, indicated the ratio of AST/ALT >2 (Table 5). Elevated activities of AST and ALT in advanced stage of alcoholic and ratio always more than 2 as compared to controls indicating that the levels was directly related to alcohol consumption and extent of the liver damage. 10,11 Some interrelated reasons have been reported for the high AST/ALT ratio in alcoholic liver cirrhosis: i) A decreased hepatic ALT activity; ii) Pyridoxal 5 phosphate depletion in the liver of alcoholics; iii) Mitochondrial damage leading to an increase in the serum activity of mitochondrial aspartate in patients with high alcohol consumption. There may also be some contribution of the direct toxic effect of alcohol on the AST/ALT ratio in addition, the estimation of the De ritis ratio is essential for the rational understanding of the extent of damage in. 12,13 Serum γgt activities were increased highly significantly both group of patients as compared to controls (p < 0.001). γ-glutamyl transferase (γgt) is a microsomal enzyme with a wide tissue distribution. It is a membrane bound glycoprotein enzyme, catalyzes the transfer of the gammaglutamyl moiety of glutathione to various peptide acceptors and this biliary canalicular enzyme is induced by alcohols and its serum level rises in response to acute hepatocellular damage. 14 The AST/ALT ratio, γgt and total bilirubin in non alcoholic correlated with AST/ALT ratio, γgt and total bilirubin in alcoholic liver (Table 4). There correlation was negatively correlated with each other. Conclusion: In this study there is comparison of diagnostic markers of in non alcoholic and alcoholic patients. Hyperbilirubinemia, hyperurea, 4
hypercreatinine, hypoalbuminemia are common features of alcoholics as well as non alcoholic liver. Monitoring γgt, ALP, AST and ALT in combination is a sensitive means of detecting severity of liver damage. All these parameters in combinations may be useful indicator for identification and determination of severity in comparison of alcoholic as well as non alcoholic s. As Maharashtra is one of the top states in name of alcohol abuse there is a need of conducting further studies in relation to this and aware general population about the various clinical effects and fatality chronic alcohol consumption may cause. Acknowledgement: The authors wish to express their deepest gratitude to all the patients and healthy volunteers who have participated in this study. References: 1. Pradhan R, Lekharu R, Srivastava R, Sharma D. A Study of Oxidative Stress in Alcoholic Liver Disease GCSMC J Med Sci 2014;3(1):16-17. 2. Joseph V, Rahman CPA. Analysis of biochemical markers in alcoholic liver cirrhosis. International Journal of Recent Trends in Science and Technology 2016;18(2):311-312. 3. Bhargava S, Singh N, Matreja PS, Gupta AK, Singh A. Radiological and Biochemical aspect of Alcoholic Liver Disease: A Prospective study. Annals of Applied Bio-Sciences 2016;3(1):2455. 4. Das BK, Lamsal M, Pradhan B, Shakya DR, Bhattarai NR, Baral N. Evaluation of biochemical parameter alteration in alcohol dependence ethnic Nepalese. International Journal of Therapeutic Applications 2014;18:1-6. 5. Saxena T, Arya A, Rathore A, Rajak N, Naz, Shah R. GGT and SGPT - A Rising Marker in Diagnosis of Non-alcoholic Fatty Liver Disease. Biomed. & Pharmacol J 2014;7(1):277-80. 6. Sass DA, Chang P, Chopra KB. Nonalcoholic fatty : A Clinical Review. Digestive Diseases and Sciences 2005;50:171-80. 7. Das SK, Vasudevan DM. Biochemical diagnosis of alcoholism. Indian Journal of Clinical Biochemistry 2005;20(1):35-42. 8. Thapa BR, Walia A. Liver function tests and their interpretation. Indian Journal of Pediatrics 2007;74:663-71. 9. Das SK, Mukherjee S, Vasudevan DM, Balakrishnan V. Comparison of haematological parameters in patients with non-alcoholic fatty and alcoholic. Singapore Med J 2011;52(3):175-81. 10. Al-Jumaily EF, Khaleel FM. The effect of chronic s on some biochemical parameters in patient s serum. Current Research Journal of Biological Sciences 2012;4(5):638-42. 11. Torkadi PP, Apte IC, Bhute AK. Biochemical evaluation of patients of alcoholic and non-alcoholic. Indian J Clin Biochem 2014;29(1):79-83. 12. Bellentani S, Saccocio G, Masutti F, Giacca M, Miglioli L, Monzoni A, et al. Risk factors for alcoholic. Addiction Biology 2000;5(3):261-8. 13. Patil AA, Katkam RV, Sawant SD. Biochemical markers in alcoholic liver cirrhosis. International Journal of Biotechnology and Biochemistry 2016;12(2):185-95. 14. D Agata ID, Balistreri WF. Evaluation of liver in the pediatric patient. Pediatrics in Review 1999;20(11):376-88. 5