| Objective:research weather the cholesterol metabolism-related genes exist DNA methylation, and find out the DNA methylation screening.Methods:There are 25 patients with gallbladder stones (GS group),35 cases of hepatolithiasis (HS group) and 25 cases of non-gallstone patients (control group), from whose liver tissue we extract DNA, Adopting Methylation specific polymerase chain reaction (methylation-specific PCR, MSP) method. We detect the methylation status of liver tissue's SRBI, CYP7A1, HCM-COA R, BSEP and other gene promoter,and then screen methylated adress of DNA. We treat it with sodium bisulfite after DNA denatured, which transform the unmethylated cytosine in the DNA into thymine, and then detect DNA methylation by PCR amplificating and agarose gel electrophoresis. The operation to obtain the gallstones, fully washed with running tap water, then distilled water rinse, place a clean gauze, and allowed themselves to dry completely. The stones set in a mortar specimens inquiry into powder, then put in the dryer, after re-drying three days and weighting them, Weigh the quality of quantitative and analyze chemically, extract Cholesterol with diethyl ether, color with Sulfuric acid color method of ferric chloride color, assay cholesterol content by spectrophotometer to determine the nature of stone。Results:SRBI gene in gallstone group which is methylated was 8.0%(2/ 25),and in the hepatolithiasis group and non-stone group were 8.0%(3/35) and 28%(7/25), comparing among the three groups, the difference was not statistically significant (X2=5.61, P> 0.05), pairwise comparisons among the three groups, bile duct stones and non-stone group, X2= 4.9, P<0.05, which is statistically significant, methylation of hepatolithiasis was lower than non-stone group of patients, comparing gallbladder stone group with non-stone group, X2=1.5, P> 0.05, the difference was not statistically significant, between the group and non-stone group of stones X2=4.1, P<0.05, statistically significant, the methylation of the stone group was lower than non-methylation stone group, the rate of methylation of BSEP in the gallbladder stone patients was 24.0%(6/25), in patients with hepatolithiasis group was 37.1%(13/35), the all stone groups was 31.7%(19/60), in non-stone group was 12.0%(3/25),among the bile duct stones, gallbladder stones and non-stone group, X=11.7, P<0.05, the differences among the three groups were statistically significant, the rate of Methylation among the three groups are different, hepatolithiasis group of patients was higher than non-stone group, pairwise comparison among the three groups, bile duct stones and non-stone group of statistical parameters X2=3.52, P> 0.05, the difference was not statistically significant. the value of statistical parameters of the BSEP methylation of stone group and non-stone group, X2=2.61, P> 0.05, the difference was not statistically significant, comparing gallstones group with the non-stone group also was non-statistical significance. HMG-COAR and CYP7A1 methylation of stones in the gallbladder, bile duct stones and non-stone groups, comparisons between any two, there were no statistical significance.Conclusion:In the process of gallstone formation, methylation of candidate gene of gallstones may be the one of the major reasons, This study found that gallstone patients show high methylation of BSEP gene, SRBI appears hypomethylation, BSEP gene hypermethylation can lead to down-regulation of its own, hypomethylation of SRBI can express their own up-regulation, Both of them can lead to the proportion of cholesterol in bile increasing, which is beneficial to form gallstone.
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