The Role of the PNPLA3, TM6SF2 and MBOAT7 Genetic Variants in the Response to Silybin-phospholipid Complex, Vitamin D and Vitamin E Based Therapy for Non-alcoholic Fatty Liver Disease Patients

The aim of this study was to evaluate if the PNPLA3 rs738409, TM6SF2 rs58542926 and MBOAT7 rs641738 could influence the response of NAFLD patients regarding metabolic parameters, liver damage and hepatic fat accumulation ones, to a treatment with 303 mg of silybin-phospholipids complex, 10 mg of vitamin D, and 15 mg of vitamin E twice a day for six months.

The investigators performed a baseline comparison of Weight, waist-to-height ratio (WHtR), blood pressure measurement, body mass index (BMI), Blood glucose and insulin, the homeostatic model for insulin resistance assessment (HOMA-IR), aspartate and alanine aminotransferase (AST, ALT), gamma-glutamyl transferase (GGT), blood count, C reactive protein (CRP), the Thiobarbituric Acid Reactive Substance (TBARS), liver stiffness and controlled attenuation parameter (CAP) among the three study groups: NAFLD wild type control group (n. 30), NAFLD treated wild type group (n. 30), NAFLD treated mutated group (n. 32). The block randomization method was used to randomize the 60 not mutated patients in the NAFLD wild-type control group and NAFLD treated wild type one, using the online randomization software http://www.graphpad.com/quickcalcs/index.cfm.

The wild-type control group was composed by NAFLD patients without PNPLA3, TM6SF2 and MBOAT7 mutations that did not receive any type of treatment during the study period.

The patients inserted in the NAFLD treated wild type and NAFLD treated mutated groups were undergone to an oral administration of 303mg of silybin-phospholipid complex, 10mg of vitamin D, and 15mg of vitamin E, twice a day for six months. None of the enrolled patients dropped-out the study.

At the end of treatment, the assessment of WHtR, blood pressure measurement, BMI, Blood glucose and insulin, HOMA-IR, AST, ALT, GGT, blood count, CRP, TBARS, liver stiffness and CAP was re-performed. During the experimental observation, patients were on free diet on the basis of dietary habits before the enrollment and any type of physical exercise was recommended during the study period. Food intake was evaluated both at baseline and end of treatment using a computerized software. The investigators recorded, with a diet diary, the food intake of a complete week, including working days and the weekend. On the basis of the quantities and qualities of food consumed, the soft elaborates the daily energy intake and the percentage/caloric amount of macronutrients. For the physical exercise assessment, the investigators submitted a specific questionnaire at baseline and end of treatment with some simple questions: Are the participant doing or have the participant ever done (in the last two years) sport in a continuative and regular way? Have the participant changed his/her daily physical activity in the last six months? 1: no, 2: yes; if yes, has the physical activity enhanced or worsened? Alcohol consumption was assessed at the beginning and at the end of the treatment. One hundred two patients with histological diagnosis of NAFLD followed by the Hepatogastroenterology Division of the University of Campania "Luigi Vanvitelli, between January and October 2017 were screened, after signing an informed consent, for the PNPLA3 rs738409, TM6SF2 rs58542926 and MBOAT7 rs641738 genetic variants and thirty-two met the inclusion criteria for the study and showed at least one among PNPLA3 I148I/M, I148M/M, TM6SF2 167E/K, 167K/K and MBOAT7 TMC4C/T or TMC4T/T genetic variants, were enrolled together with sixty patients without the mutations. Ten patients were excluded from the enrollment due to the coexistence of several comorbidities and/or advanced stages liver disease such as cirrhosis and/or hepatocellular carcinoma (HCC).