Study On UGT1A1 Gene Polymorphism And The Relationship Of Primary Intrahepatic Bile Duct Stones

Objective: These research studied the releationship between UGT1A1 gene polymorphism and the formation of Primary Intrahepatic Stones PIS by analyzed the bilirubin metabolism releatedgene polymorphism which included UGT1A1*28 and UGT1A1*6. For the prevention and treatment of Primary Intrahepatic Stones in the certain theory basis.Subjects and Methods:1.the subjectsSelecting October 2013 一 May 2014, the people’s hospital of qingzhou of primary intrahepatic biliary calculi patients as the experimental group, in total of thirty-two cases. Of which eighteen cases of patients with intrahepatic bile duct stones left,the right hepatic duct stones were ten cases, about four cases of patients with intrahepatic bile duct stones; twenty males and twelve females; at the age of 28-76 years old, average age is 52.68 years old.Choose fifteen healthy subjects as control group, ten cases of male, five cases of female; at the age of 20-72 years old, average age is 46.72 years old. All control patients by color Doppler ultrasound and abdominal CT examination showed no bile duct stones.Extracting object in experiment fasting fresh whole blood 10 ml, divided into three equal parts which is arranged in the containing anticoagulant negative pressure pumpingblood vessels, upside down mixing, one sending to laboratory in our hospital, determination of serum bilirubin level, the other two copies are stored in the-70 degrees Celsiusfreezer.2 experimental methodsUsing DNA extraction kit, extraction experiment object vein blood DNA, using the method of RT-PCR, the promoter region of UGT1A1 gene(*28 locus) and the first exon region(site *6)sequence of gene amplification and sequencing, analysis the genotype ofsample. Expression of B-UGTm RNA was detected by RT-PCR in blood. To analyze the relationship between UGT1A1*28and UGT1A1*6 with primary bile duct stone formation in liver by statistical method.Results:1. The distribution of UGT1A1*28 gene and UGT1A1*6 gene frequencies in the groups of Intraccholedocholithiasis and controls.1.1 intrahepatic calculi group UGT1A1* 28 gene frequency distribution: the wildtype(*1/*1) 87.5%(n=28), the heterozygous mutant(*1/*28) was 12.5%(n=4),the homozygous mutant(*28/*28) was 0(n=0).The distribution of UGT1A1*6 gene frequencies in the groups of Intraccholedocholithiasis : The percentage of wild type(G/G) is 68.7%(n=22),The heterozygous mutant(G/A) is 12.5%(n=4),The homozygous mutant(A/A)is 6.3%(n=2).1.2 The distribution of UGT1A1*28 gene frequencies in the groups of control : The percentage of wild type(*1/* 1) is 60%(n=9),The heterozygous mutant(*1/*28) is40%(n=6),The homozygous mutant(*28/*28) is 0(n=0).The distribution of UGT1A1*6 gene frequencies in the groups of control : The percentage of wild type(G/G) is 100%(n=15), Not found the mutation genotype.2.The comparison of the distribution of UGT1A1*28 gene and UGT1A1*6 gene frequencies in the groups of Intraccholedocholithiasis and controls:there is no significance of UGT1A1*28 between two groups( χ2=3.115, P > 0.05),but the expression of UGT1A1* 6 gene in Intraccholedocholithiasis group is significantly higher than the control.3.The mean expression of B-UGTm RNA in the biood of Intraccholedocholithiasis patients is 1.059±0.014,the control is 2.635±0.106,so the former is signicantly lower than the latter(P<0.05).Conclusion:1. UGT1A1 * 28 and the formation of primary intrahepatic biliary calculi has no obvious relation.2. UGT1A1 * 6 is probably one of the risk factors of primary intrahepatic bile duct stone formation. But due to the small number of cases,the area is limited,need to expand the clinical cases, further validation.