A Study Of The Relationship Between The Expression Of Bilirubin UDP-glucuronsytransferase1A1and Primary Intarhepatic Stones

Objective:Bilirubin UDP-glucuronsytransferase(B-UGT) is the key enzyme in bilirubin metabolism,its coding gene is UGT1A1.The previous reports indicated that the reduction of hepatic activity of bilirubin UDP-glucuronsytransferase might associated with the increase of monoconjugated bilirubin(MCB) in bile with which leads to lithogenic bile. this experiments study the pathogenesis of primary intrahepatic stones by measuring the expression of UGT1A1of mRNA and protein, to explore the relationship between the expression and regulation in primary intrahepatic stones.Observation and Methods:1、There are33patients (19males,14females) with primary intrahepatic stones(HS group) and15non-primary intrahepatic stones(NS group:9males,6females), which consisted of8hepatic cysts,6hepatic hemangiomas,1liver cancer. Each primary intrahepatic stones were identified to be pigments stone in surgery. The liver tissues were frozen eighty degrees below zero immediately in central laboratory after the sugery.2、Measure the expression of UGT1A1mRNA:the expression of UGT1A1mRNA was analysed by real-time PCR(SYBR Green relative quatitative), GDPH is used as house-keeping gene. The forward primer of real-time PCR is TGCCCACTGTATTC TTCTTGC(5’-3’) and reverse primer is TTCTGTGAAAAGGC AATGAGC (5’-3’).The expression of mRNA is analysed by Bio-Rad iQ5according to the standard curve and Ct value.3、Measure the expression of UGT1A1protein:the expression of bilirubin UDP-glucuronsytransferase proteins was analysed by western blot, B-action is used as house-keeping gene.The contents of total cholesterol(TC),total bile acid(TBA),Triglyceride(TG), low density lipoprotein(LDL),and high density lipoprotein(HDL),total bilirubin(TB), direct bilirubin(DB),indirect bilirubin(IDB) were measured in preoperative plasma with empty stomch.Results:1、analysis of the plasma:The total cholesterol,total bile acid, triglyceride, low density lipoprotein, and high density lipoprotein, total bilirubin, direct bilirubin, indirect bilirubin in non-primary intrahepatic stones group(NS) is higher than primary intrahepatic stones group(HS group). But the difference is not statistically significant.2、The mRNA expression of UGT1A1in primary intrahepatic stones group were increased obviously than non-stones group(10.246±2.151vs.1.000±0.242),(t=2.89,p=0.018).3、 The proteins expression of UGT1A1in primary intrahepatic stones group were higher than non-stones groups(1.059±0.232vs.0.835±0.007),(t=2.487,p=0.020).3、there were no correlations between the mRNA and protein(B-UGT) expressions of UGT1A1gene (r=-0.039, P=0.827).There are zero correlation between the mRNA、protein(B-UGT) expressions of UGT1A1gene and total cholesterol,total bile acid,triglyceride, low density lipoprotein, and high density lipopro,total bilirubin, direct bilirubin, indirect bilirubin.Conclusion:The UGT1A1gene might be associated with primary intrahepatic stones.The mRNA and proteins expression of UGT1A1were increased obviously in primary intrahepatic stones. The expression of UGT1A1gene maybe not correlated with the decreased activity of hepatic B-UGT.