Molecular characterization of patatin like phospholipase domain containing 3 (pnpla3)gene in alcoholic liver disease

Abstract

BACKGROUND Alcoholic and non-alcoholic fatty liver disease (ALD and NAFLD), or alcoholic and non- alcoholic fatty liver disorders, have grown to be major public health concerns in contemporary nations. Global alcohol consumption was estimated to be 6.18 litres per person on average. Hepatitis, cirrhosis, hepatocellular cancer, and steatosis are the symptoms of alcoholic liver disease (ALD). However, only 6–41% of heavy drinkers who consume more than 60–80 g/day of alcohol (and more than 20 g/day for women) go on to develop cirrhosis. Along with those environmental factors, a number of genetic variants have been linked to ALD and NAFLD. PNPLA3 is a lipoatrophic protein that is a member of the PNPLA family and has 481 amino acids. The relationship of PNPLA3(148M) with a wide range of liver illnesses, including cirrhosis, fibrosis, non-alcoholic steatohepatitis (NASH), ALD and NAFLD, and hepatocellular carcinoma (HCC), has now been confirmed by numerous genetic investigations.The underlying pathogenic mechanisms, however, are still unknown.1 .As few studies were done on the Indian population so this study is carried out to find out the Molecular Characterization of patatin like phospholipase domain containing 3(PNPLA3) gene in alcoholic liver disease, AIMS AND OBJECTIVES Study of Molecular characterization of Patatin like phospholipase domain containing 3 (PNPLA3) gene in alcoholic liver disease. MATERIALS AND METHODS. It is a cross sectional study done on a Patients admitted with Alcoholic liver disease at BLDE (Deemed to Be University) Shri B M Patil Medical College Hospital and Research Centre, Vijayapura, during the period from May 2023 to Dec 2024.The sample size is 60. All Alcoholic Liver Disease diagnosed by clinical-history and examination, radiological- ultrasonography and biochemical parameters- like LFT(AST/ALT). METHODOLOGY A commercially available DNA kit is used to separate genomic DNA from 300μl of peripheral blood. PCR amplification is performed in a 20μl reaction volume with 0.5μl of each primer (5pmol) and 0.5μl of genomic DNA (75ng/μl to 150 ng/μl). The primer's annealing temperature is determined by annealing it for 10 seconds at 72°C for 154 seconds (primer extension) , followed by the final extension at 72°C which is for 5 minutes. PCR products are verified for their corresponding amplicon size using a standard 100bp ladder in gel electrophoresis.

Statistical Analysis: After the data is gathered and imported into a Microsoft Excel sheet, statistical analyses are performed using SPSS (Statistical Programme for the Four Social Sciences, Version 20). The results are shown as mean, SD, counts, percentages, and graphs. Four continuous variables, each normally distributed, will be compared between the two groups using an independent sample test. When dealing with non-normally distributed variables, the Mann-Whitney U test will be employed. To compare categorical variables between the two groups, the chi-square test or Fisher's exact test is employed. RESULTS We found that out of 60 patients , 25 patients (i.e,41.7%) were aged between 41-50 years , followed by 19 patients (31.7%) were between 31-40 years. The mean age and Standard deviation is 42.58 and 8.7 respectively.58 pateitns were Males (96.7%) and 2 were females(3.3%). Majority consumed alcohol for a duration of 11 to 20 years i.e, 35 patietns (58.3%) and followed by for <10 years 19 patients 31.7% .Mean and Standard Deviation of Various parameters like BMI, TB, CB, UB, Albumin, Globulin is 22.0(3.31), 7.23(8.56),5.30(7.75),1.90(1.56),2.6(0.8), 3.4(0.818) respectively. 49 patients (81.7%) had no mutation recorded and 5 patients (8.3%) had rs738409 gene positive and 6 patients(10%) had rs738409, rs738408 genes positive . Out of 60 patients , 11 (18.3%)had the gene mutation, and 49 (81.7%) did not have any mutation. However, this gene is not related to age, gender, years of alcoholic consumption and stage of the disease (p-value>0.05).

CONCLUSION: This study shows a clear link between certain PNPLA3 gene variations and Alcoholic Liver Disease, especially the rs738409 variant, which seems to be tied to more severe cases. We found that in our study there is no statistically significant between the demographic details, stage of the disease, and years of alcohol consumption.