Ignite Creation Date:
2024-05-06 @ 7:08 PM
Last Modification Date:
2024-10-26 @ 3:01 PM
Study NCT ID:
NCT05905185
Status:
COMPLETED
Last Update Posted:
2023-06-15
First Post:
2023-05-01
Brief Title:
Interventional Strategy in Tackling Emerging Non-alcoholic Fatty Liver Disease in Childhood Obesity
Sponsor:
National University of Malaysia
Organization:
National University of Malaysia
Study Overview
Official Title:
Interventional Strategy in Tackling Emerging Non-alcoholic Fatty Liver Disease in Childhood Obesity
Status:
COMPLETED
Status Verified Date:
2023-06
Last Known Status:
None
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
False
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
None
Brief Summary:
The goal of this clinical trial is to investigate the effects of tocotrienol-rich fraction vitamin E supplementation on liver enzymes in overweight and obese children with non-alcoholic fatty liver disease as compared to placebo
The main questions it aims to answer are
1 Does supplementation of tocotrienol-rich fraction vitamin E reduce the level of liver enzymes and improve liver steatosis in non-alcoholic fatty liver disease among overweight and obese children
2 Does tocotrienol-rich fraction vitamin E supplementation improve the level of liver steatosis by reducing the level of DNA damage
Participants will
1 consume daily either a dose of 50 mg of tocotrienol-rich fraction TRF vitamin E or a placebo for 6 months
2 Routine clinical assessments include weight height waist circumference and BMI Fasting glucose and fasting serum lipid
3 The following investigations were performed upon recruitment and following 6 months of intervention i liver biomarker and enzymes ii DNA damage iii TNFα IL-6 and IFN-gamma genes iv Fibroscan
The main questions it aims to answer are
1 Does supplementation of tocotrienol-rich fraction vitamin E reduce the level of liver enzymes and improve liver steatosis in non-alcoholic fatty liver disease among overweight and obese children
2 Does tocotrienol-rich fraction vitamin E supplementation improve the level of liver steatosis by reducing the level of DNA damage
Participants will
1 consume daily either a dose of 50 mg of tocotrienol-rich fraction TRF vitamin E or a placebo for 6 months
2 Routine clinical assessments include weight height waist circumference and BMI Fasting glucose and fasting serum lipid
3 The following investigations were performed upon recruitment and following 6 months of intervention i liver biomarker and enzymes ii DNA damage iii TNFα IL-6 and IFN-gamma genes iv Fibroscan
Detailed Description:
Methodology This study enrolled a diverse group of participants aged 10 to 18 regardless of gender who were diagnosed with fatty liver disease detected via ultrasonography and abnormally high alanine transaminase levels at least two-fold higher than the upper limits for their respective genders The trial consisted of 29 patients with 15 receiving a daily oral dose of 50 mg TRF and 14 receiving a placebo for a duration of six months Various clinical parameters LIVERFASt in vitro diagnostic test fibroscan biochemical parameters DNA damage and gene expression both at the outset and at the end of the study were monitored
The amount of blood sample taken was 13mls each at entry and on completion at 6 months Details of these tests are as shown below
Transient Elastography FibroScan
This is a non-invasive test to measure liver stiffness kPa indicator of liver fibrosis and to detect degree of fat accumulation CAP range value of 100-400 dBm liver stiffness of 8kPa indicates advanced fibrosis while CAP of 263 indicates fatty liver During the screening the participant lied down on the examination bed with hisher right hand behind hisher head A probe was placed by the investigator between the right ribs of the patient Measurements were recorded into the machine FibroScan 502 Touch The screening is quick and easy usually taking less than 15 minutes The scanned steatosis was scored and graded
Determination of DNA Damage The DNA Damage pre and post intervention were conducted by using CometAssay Kit Trevigen Gaithersburg USA following the manufacturers protocol Briefly blood cells in 5 μL medium was suspended in 70 μL warm 06 low-melting point LMA agarose Boehringer Mannheim Germany DNAse-free RNAsefree Slides were allowed to sit in the alkaline buffer for 20 min to allow unwinding of DNA strands and expression of alkali-labile damage Electrophoresis was performed for 20 min at 300 mA and 25 V Following dropwise neutralization TrisHCl pH 75 for 5 min cells were stained by applying 30 μL 1X ethidium bromide The slides were examined and the tail length was measured with a fluorescence microscope Carl Zeiss Germany The DNA migration of 100 randomly selected cells were examined for each sample A total damage score was determined by multiplying the number of cells assigned to each grade of damage by the numeric value of the grade according to methods described by Heaton et al 19 Total DNA damage score was calculated as follows Total DNA damage 0 n0 1 n1 2 n2 3 n3 4 n4 where n0 cells with Score 0 n1 cells with Score 1 n2 cells with Score 2 n3 cells with Score 3 and n4 cells with Score 4
Determination of liver biomarker and enzyme The levels of liver enzymes including alanine aminotransferase ALT aspartate aminotransferase AST gamma glutamyl transferase GGT and alkaline phosphatase ALP pre and post intervention were determined from the blood samples of the patients Other biomarkers such as lipid profile α-2 macroglobulin and haptoglobin were also be analysed The blood samples were collected in a BD Vacutainer and centrifuge at 1500 xg for 15 mins for the serum separation The serum was stored in -80C if it is not processed immediately The levels of the liver enzymes were determined by using ADVIA 1800 Siemens USA
Quantitative Polymerase Chain Reaction qPCR
Total RNA was extracted from blood samples of NAFLD patients with or without tocotrienol Primers and probes for TNFα IL-6 and IFN-gamma genes were designed 1 ugul of total RNA was converted to cDNA which then be used for PCR reaction All samples were analysed in duplicate Expression of genes was determined after normalised with the housekeeping genes
The amount of blood sample taken was 13mls each at entry and on completion at 6 months Details of these tests are as shown below
Transient Elastography FibroScan
This is a non-invasive test to measure liver stiffness kPa indicator of liver fibrosis and to detect degree of fat accumulation CAP range value of 100-400 dBm liver stiffness of 8kPa indicates advanced fibrosis while CAP of 263 indicates fatty liver During the screening the participant lied down on the examination bed with hisher right hand behind hisher head A probe was placed by the investigator between the right ribs of the patient Measurements were recorded into the machine FibroScan 502 Touch The screening is quick and easy usually taking less than 15 minutes The scanned steatosis was scored and graded
Determination of DNA Damage The DNA Damage pre and post intervention were conducted by using CometAssay Kit Trevigen Gaithersburg USA following the manufacturers protocol Briefly blood cells in 5 μL medium was suspended in 70 μL warm 06 low-melting point LMA agarose Boehringer Mannheim Germany DNAse-free RNAsefree Slides were allowed to sit in the alkaline buffer for 20 min to allow unwinding of DNA strands and expression of alkali-labile damage Electrophoresis was performed for 20 min at 300 mA and 25 V Following dropwise neutralization TrisHCl pH 75 for 5 min cells were stained by applying 30 μL 1X ethidium bromide The slides were examined and the tail length was measured with a fluorescence microscope Carl Zeiss Germany The DNA migration of 100 randomly selected cells were examined for each sample A total damage score was determined by multiplying the number of cells assigned to each grade of damage by the numeric value of the grade according to methods described by Heaton et al 19 Total DNA damage score was calculated as follows Total DNA damage 0 n0 1 n1 2 n2 3 n3 4 n4 where n0 cells with Score 0 n1 cells with Score 1 n2 cells with Score 2 n3 cells with Score 3 and n4 cells with Score 4
Determination of liver biomarker and enzyme The levels of liver enzymes including alanine aminotransferase ALT aspartate aminotransferase AST gamma glutamyl transferase GGT and alkaline phosphatase ALP pre and post intervention were determined from the blood samples of the patients Other biomarkers such as lipid profile α-2 macroglobulin and haptoglobin were also be analysed The blood samples were collected in a BD Vacutainer and centrifuge at 1500 xg for 15 mins for the serum separation The serum was stored in -80C if it is not processed immediately The levels of the liver enzymes were determined by using ADVIA 1800 Siemens USA
Quantitative Polymerase Chain Reaction qPCR
Total RNA was extracted from blood samples of NAFLD patients with or without tocotrienol Primers and probes for TNFα IL-6 and IFN-gamma genes were designed 1 ugul of total RNA was converted to cDNA which then be used for PCR reaction All samples were analysed in duplicate Expression of genes was determined after normalised with the housekeeping genes
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
False
Is a FDA Regulated Device?:
False
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None