Objective To investigate the effects of activities of Na+K+ATPase in plasma membranes of hepatocytes on the formation of calcium bilirubinate gallstone. MethodsThe research were studied in Xiao’s rabbit models.One hundred and three rabbits were randomly divided into the control group (Con,n=28),simple biliary obstruction group (BO,n=36),biliary obstruction and infection group (BOI,n=39).The activities of Na+K+ATPase,contents of intracellular calcium and bile acids in bile were measured in 3rd,7th,14th and 20th day. ResultsThe activities of Na+K+ATPase,contents of bile acids in bile decreased gradually and the contents of intracelluar calcium of hepatocytes increased continously in all phases of experimental groups compared with control group (P＜0.01).There was more significant changes in BOI group than in BO group (P＜0.05). Conclusion The progressive decrease of activities of Na+K+ATPase has a close relationship with the overloaded intracellular calcium and the continous decrease of bile acids contents in bile in formation of calcium bilirubinate gallstone.Infection can promote above mentioned changes and more stone formation.
To study of plasma lipoprotein cholesterol and effects of these changes on bile acids and cholesterol in bile during gallstone formation in rabbit model. This gallstone model was induced by high cholesterol diet (HCD). The rabbits were divided into five groups and there were ten animals in each group. The plasma highdensity lipoprotein cholesterol (HDL-C) and its subgroups (HDL2-C, HDL3-C), low-density lipoprotein cholesterol (LDL-C), very low-density lipoprotein cholesterol (VLDL-C), total cholesterol, triglyceride, phospholipid, bile acids and cholesterol of bile were investigated in different time. The results were as follow: ①As the time of feeding HCD passed by, the plasma total cholesterol, LDL-C and VLDL-C increased markedly (3-week group and 4-week group vs control group, Ｐ＜0.05). Though the plasma HDL-C and its subfractions HDL2-C and HDL3-C did not change significantly, the function of HDL in transporting plasma cholesterol decreased markedly (from 80.00% to merely 3.68%); ②Cholesterol in bile increased gradually and there were significant differences when 3-week group and 4-week group comparing with control group. The concentration of GDCA and GCA in bile changed slightly (Ｐ＞0.05). These results suggest that the changes of plasma lipoprotein cholesterol may affect the metabolism of cholesterol and bile acids and it may take an important role in the formation of gallstone.
The aim of the this study was to search for bacterial DNA sequences in cholesterol gallstones with negative bacterial culture by NP-PCR technique. Bacterial gene fragments were amplified in vitro from DNA which were extracted from cholesterol gallstones in gallbladder for identifying the existence of bacteria. The gallbladder gallstones of 30 patients were analysed. Bacterial DNA was found in the stones of 26 patients, indicating that most cholesterol gallstones harbor bacterial DNA.
Objective To discuss the changes of c-kit/scf mRNA and protein in guinea pig gallbladder fed on high cholesterol diet. Methods Twenty guinea pigs were divided into two equal groups of 10 each：the control group and lithogenic group. Normal diet and high cholesterol diet was given to each group respectively. The period of stone permeation was six weeks. RT-PCR and Western blot were used to determin the expressions of c-kit and scf mRNA and protein. Results RT-PCR results showed that the expressions of c-kit mRNA（t=6.985，P＜0.01） and scf mRNA （t=6.028， P＜0.01）decreased significantly in lithogenic group compared with the control group. Western blot results showed that the expressions of c-kit protein （t=10.256， P＜0.01） and scf protein （t=9.586， P＜0.01）decreased significantly in lithogenic group compared with the control group. Conclusions The expressions of c-kit/scf mRNA and protein decrease during the formation of cholesterol gallstones in guinea pigs fed on high cholesterol diet. Inhibition of c-kit/scf pathway may play a role in the formation of cholesterol gallstones.
Objective To review the mechanisms of cholesterol gallstones caused by female hormone so as to explore new treatments to prevent gallstones associated with estrogen and progesterone. Methods The literatures on gallstones related with female hormone were reviewed and the mechanisms of cholesterol gallstones were summarized. Results The cholesterol gallstones mechanisms was affected by estrogen through genomic effects，and the nucleation of cholesterol was promoted by estrogen through nongenomic，which resulted in the formation of cholesterol gallstones. And the bile empty dysfunction associated with estrogen through nongenomic effects was also the reason of cholesterol gallstone formation. The G proteins α subunit responsible for the motility of gallbladder were disrupted by progesterone through genomic effects，and the ionic channels and signal transduction were also interfered through nongenomic pathway，which impaired the contraction of gallbladder. However，the nongenomic effects might not play an important role in the gallstones formation caused by progesterone. Conclusions The mechanisms of cholesterol gallstones formation associated with female hormone are complicated，the understanding of chelesterol gallstones formation mechanisms might be helpful to prevent gallstones associated with estrogen and progesterone.
Objective To investigate the mRNA expressions of liver X receptor α (LXRα), farnesoid X receptor (FXR), steroid xenobiotic receptor (SXR) and liver receptor homolog 1 (LRH-1) gene in patients with cholesterol gallstone (CGS) disease in order to elucidate the biomolecular pathogenesis of gallstone formation. Methods Twenty-seven patients with CGS (CGS group) and 10 controls without gallstones (control group) were included in this study. Serum lipid composition (total cholesterol, triglyceride, high density lipoprotein cholesterol, apoprotein B, apoprotein A1), gallstone cholesterol concentration and biliary composition (cholesterol, bile salts, lecithin) were assayed. Biliary total lipid and cholesterol saturation index (CSI) were calculated. mRNA expressions of LRH-1, FXR, SXR and LXRα gene were determined by real-time polymorphism chain reaction. Results Serum high density lipoprotein cholesterol concentration was lower in CGS group than that in control group 〔(0.93±0.05) mmol/L vs (1.33±0.09) mmol/L, P＜0.001〕 and serum apoprotein A1 was also lower in CGS group than that in control group 〔(1.19±0.05) g/L vs (1.36±0.06) g/L, P＜0.05〕. There were no differences in serum total cholesterol, triglyceride and apoprotein B between two groups (Pgt;0.05). CSI was higher in CGS group than that in control group (1.17±0.02 vs 0.79±0.10), P＜0.001. Biliary cholesterol was also higher in CGS group than that in control group 〔(7.96±0.39) mol% vs (5.26±0.89) mol%, P＜0.01〕, while biliary total lipid was lower in CGS group than that in control group 〔(104.72±10.51) g/L vs (154.24±14.20) g/L, P＜0.05〕. There were no differences in bile salts and lecithin between two groups (Pgt;0.05). Expression of LRH-1 gene was higher in CGS group than that in control group (14.18±1.80 vs 7.22±2.22), the difference was statistically significant (P＜0.05). There were no differences in mRNA expressions of LXRα, FXR and SXR gene between two groups (Pgt;0.05). Conclusion CGS disease may be related to increased expression of LRH-1 gene.
Objective In order to study the mechanism of cholesterol gallstone formation through rabbit model which was induced by high cholesterol diet (HCD)Methods the activities of the high density lipoprotein receptor (HDLR) and low density lipoprotein receptor (LDLR) of hepatocytes were investigated. Results The results were as follows: The HDLR activity increased significantly after taking HCD for one week, at the same time, the LDLR activity only increased slightly. Thereafter, the activities of HDLR and LDLR all decreased markedly. As the time of animals taking HCD went on, serum total cholesterol, LDL cholesterol and hepatic cholesterol increased, but bile acids of biliary tract decreased gradually. Conclusion The results suggest that the changes of HDLR and LDLR activities of hepatocytes had no significant effect on bile cholesterol and the decreased HDLR and LDLR activities may cause the reduction some of substrate for bile acids synthesise and play an important role in the formation of gallstone.
ObjectiveTo explore the expressions of transthyretin(TTR) mRNA and its protein in tissues of human gallbladder with cholesterol gallstones, and to explore its role in the formation of cholesterol gallstones. MethodsGallbladder were got from cases of cholesterol gallstones(cholesterol gallstones group, n=25) and cases underwent liver transplantation with normal gallbladder(normal control group, n=9) respectively, who were treated in Ren Ji Hospital and Huashan Hospital. Real time PCR(RT-PCR) and Western blot method were used to determine the expressions of TTR mRNA and its protein respectively. In addition, 2 kinds of artificial model bile system were established to test nucleation time(NT) and nucleation activity, which added TTR and albumin(ALB). ResultsThe expression levels of TTR mRNA and protein in cholesterol gallstones group were 1.51±0.78 and 3.95±0.09 respectively, which were both higher than those of normal control group(P<0.05). The NT were(14.5±1.3)d and(18.0±0.8)d in TTR group and ALB group in small model bile system(P<0.01), which was similar with comprehensive model bile system[(13.5±0.6)d vs. (18.5±1.3)d]. The nucleation activity were 0.81 and 0.73 in small model bile system and comprehensive model bile system respectively. ConclusionsExpression of TTR up-regulates in human gallbladder tissues of patients with cholesterol gallstones, and TTR plays role of nucleation in model bile system, which is related to the formation of cholesterol gallstones.
Objective To explore association of apolipoprotein B (ApoB) gene rs676210 and rs2854725polymorphisms with gallstone disease and differences of polymorphisms between Uygur population and Han population. Methods A case control study was used. One hundred and eighty-nine patients with gallstone disease from 2010 to 2014 in our hospital were collected, of which 99 cases of Uygur population and 90 cases of Han population. One hundred and ninety age- and sex-matched healthy volunteer accepted physical examination in our hospital over the same period were collected as control, of which 93 Uygur population and 97 Han population. The ApoB genotyping of DNA samples were amplified by using SNaPshot single nucleotide polymorphism (SNP). The differences of polymorphisms between Uygur population and Han population and between patients with gallstone disease and healthy volunteer were analyzed. Results ① The differences of ApoB gene rs676210 and rs2854725 allele frequencies were not found between the patients with gallstone disease and healthy volunteer whether Uygur population or Han population (Uygur population: rs676210:χ2=0.229,P=0.633; rs2854725:χ2=0.028,P=0.866. Han population: rs676210:χ2=0.608,P=0.435; rs2854725:χ2=2.673,P=0.102). ② The differences of ApoB gene rs676210 and rs2854725 allele frequencies were not found between Uygur population and Han population whether the patients with gallstone disease or healthy volunteer (Patients with gallstone disease: rs676210:χ2=0.103,P=0.748; rs2854725:χ2=3.139,P=0.076. Healthy volunteer: rs676210:χ2=0.000,P=0.990; rs2854725:χ2=2.673,P=0.102). ③ The differences of ApoB gene rs676210 and rs2854725 genotype frequencies were not found between the patients with gallstone disease and healthy volunteer whether Uygur population or Han population (Uygur population: rs676210:χ2=2.301,P=0.317; rs2854725:χ2=3.040,P=0.219. Han population: rs676210:χ2=4.909,P=0.086; rs2854725:χ2=0.107,P=0.744). ④ The differences of ApoB gene rs676210 and rs2854725 genotype frequencies were not found between Uygur population and Han population patients with gallstone disease (rs676210:χ2=0.235,P=0.899; rs2854725:χ2=3.630,P=0.057). The difference of ApoB gene rs676210 genotype frequency was not found between Uygur population and Han population with healthy volunteer (χ2=1.026,P=0.599). While the difference of ApoB gene rs2854725 genotype frequency was found between Han population and Uygur population with healthy volunteer (χ2=9.153,P=0.010). When it was compared in pairs,α=0.05/3=0.016, the difference of G/T and T/T frequencies was found between Uygur population and Han population (χ2=6.128,P=0.013), G/T of Han population (27.8%) was higher than that of Uygur population (12.9%). Conclusions ApoB gene rs676210 and rs2854725 polymorphisms are not associated with gallstone disease. For healthy volunteer, ApoB gene rs676210 polymorphism shows no ethnics-specific difference between Uygur population and Han population, but ApoB gene rs2854725 polymorphism mightbe show a difference between Uygur population and Han population. Larger sample sizes and multicenter study are needed to confirm it.
ObjectiveTo explore the differential expressed lncRNA genes associated with formation of cholesterol gallstone, and analyze the biological functions of differential expressed lncRNA through bioinformatics.MethodsA total of 24 C57BL/6 mice were randomly divided into normal control group (n=8) and lithogenic group (n=16), which were treated with chow diets and lithogenic diets respectively for 5 weeks. After 5 weeks, mice of the lithogenic group were randomly divided into model control group (n=8) and ursodeoxycholic acid treatment group (n=8). Afterwards, mice of the normal control group were still fed with chow diets, mice of the model control group were fed with lithogenic diets, mice of the ursodeoxycholic acid treatment group were fed with ursodeoxycholic acid. After 2 weeks, collected liver tissues and gallbladder bile from the three groups, and observed gallbladder gross sample and analyzed lipids component of gallbladder bile, meanwhile detected the differential expressed lncRNA and analyzed the biological functions of differential expressed lncRNA through bioinformatics, including Gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) pathway analysis.ResultsWe successfully constructed the mice model of cholesterol gallstone. Total cholesterol level of gallbladder in the model control group had significantly higher than those of the normal control group and ursodeoxycholic acid treatment group (P<0.05), yet there was no significant difference between the normal control group and ursodeoxycholic acid treatment group (P=0.59). The levels of total bile acid, total bilirubin, and direct bilirubin had no significant difference among the three groups (P>0.05). There were 49 kinds of common overlapped difference lncRNA between the ursodeoxycholic acid treatment group and the model control group through differential expression analysis of lncRNA in liver tissues of the mice in three groups. GO and KEGG path analysis were performed separately by differential expressed lncRNA, and 88 kinds of GO terms and 18 kinds of pathways were significantly enriched from the model control group and the normal control group, 205 kinds of GO terms and 20 kinds of pathways were significantly enriched from the ursodeoxycholic acid treatment group and the normal control group.ConclusionsUrsodeoxycholic acid has therapeutic effect for cholesterol gallstone. Differential expressed lncRNAs play an important regulatory role in the formation of cholesterol gallstone and the prevention of gallstone formation by ursodeoxycholic acid treatment, which further lay the foundation in discussing specific mechanism regulated by lncRNA.