Effects of sphincter of Oddi motility on the formation of cholesterol gallstones

AIM: To investigate the mechanisms and effects of sphincter of Oddi(SO) motility on cholesterol gallbladder stone formation in guinea pigs.METHODS: Thirty-four adult male Hartley guinea pigs were divided randomly into two groups, the control group(n = 10) and the cholesterol gallstone group(n = 24), which was sequentially divided into four subgroups with six guinea pigs each according to time of sacrifice. The guinea pigs in the cholesterol gallstone group were fed a cholesterol lithogenic diet and sacrificed after 3, 6, 9, and 12 wk. SO manometry and recording of myoelectric activity were obtained by a multifunctional physiograph at each stage. Cholecystokinin-A receptor(CCKAR) expression levels in SO smooth muscle were detected by quantitative real-time PCR(q RT-PCR) and serum vasoactive intestinal peptide(VIP), gastrin, and cholecystokinin octapeptide(CCK-8) were detected by enzyme-linked immunosorbent assay at each stage in the process of cholesterol gallstone formation.RESULTS: The gallstone formation rate was 0%, 0%, 16.7%, and 83.3% in the 3, 6, 9, and 12 wk groups, respectively. The frequency of myoelectric activity in the 9 wk group, the amplitude of myoelectric activity in the 9 and 12 wk groups, and the amplitude and the frequency of SO in the 9 wk group were all significantly decreased compared to the control group. The SO basal pressure and common bile duct pressure increased markedly in the 12 wk group, and the CCKAR expression levels increased in the 6 and 12 wk groups compared to the control group. Serum VIP was elevated significantly in the 9 and 12 wk groups and gastrin decreased significantly in the 3 and 9 wk groups. There was no difference in serum CCK-8 between the groups.CONCLUSION: A cholesterol gallstone-causing diet can induce SO dysfunction. The increasing tension of the SO along with its decreasing activity may play an important role in cholesterol gallstone formation. Expression changes of CCKAR in SO smooth muscle and serum VIP and CCK-8 may be important causes of SO dysfunction.

Association of caveolin-3 and cholecystokinin A receptor with cholesterol gallstone disease in mice

AIM: To investigate the role of caveolin-3 (CAV3) and cholecystokinin A receptor (CCKAR) in cholesterol gallstone disease (CGD).

Biliary microbiota and mucin 4 impact the calcification of cholesterol gallstones

Background:Cholesterol gallstones account for over 80%of gallstones,and the pathogenesis of gallstone formation involves genetic and environmental factors.However,data on the evolution of cholesterol gallstones with various densities are limited.This study aimed to determine the roles of microbiota and mucins on the formation of calcified cholesterol gallstones in patients with cholelithiasis.Methods:Paired gallbladder tissues and bile specimens were obtained from cholelithiasis patients who were categorized into the isodense group and calcified group according to the density of gallstones.The relative abundance of microbiota in gallbladder tissues was detected.Immunohistochemistry and enzyme-linked immunosorbent assay were performed to detect the expression levels of MUC1,MUC2,MUC3a,MUC3b,MUC4,MUC5ac and MUC5b in gallbladder tissues and bile.The correlation of microbiota abundance with MUC4 expression was evaluated by linear regression.Results:A total of 23 patients with gallbladder stones were included.The density of gallstones in the isodense group was significantly lower than that of the calcified group(34.20±1.50 vs.109.40±3.84 HU,P<0.0001).Compared to the isodense group,the calcified group showed a higher abundance of gram-positive bacteria at the fundus,in the body and neck of gallbladder tissues.The concentrations of MUC1,MUC2,MUC3a,MUC3b,MUC5ac and MUC5b in the epithelial cells of gallbladder tissues showed no difference between the two groups,while the concentrations of MUC4 were significantly higher in the calcified group than that in the isodense group at the fundus(15.49±0.69 vs.10.23±0.54 ng/mL,P<0.05),in the body(14.54±0.94 vs.11.87±0.85 ng/mL,P<0.05)as well as in the neck(14.77±1.04 vs.10.85±0.72 ng/mL,P<0.05)of gallbladder tissues.Moreover,the abundance of bacteria was positively correlated with the expression of MUC4(r=0.569,P<0.05)in the calcified group.Conclusions:This study showed the potential clinical relevance among biliary microbiota,mucins and calcified gallstones in patients with gallstones.Gram-positive microbiota and MUC4 may be positively associated with the calcification of cholesterol gallstones.

Effect of intestinal transit on the formation of cholesterol gallstones in hamsters

BACKGROUND: The effect of 'intestinal transit' has become a new field of interest in the study of the pathogenesis of cholesterol gallstones. This study was undertaken to further test this notion and ascertain the relationship between impaired intestinal transit function and cholesterol gallstones. METHODS: A total of 64 hamsters were divided into 2 groups, experimental and control. Each was subdivided into 4 subgroups for sacrifice at different time. A high-cholesterol diet and a standard diet were fed to each group. The geometric center, which represents the intestinal transit function was calculated. RESULTS: The growth of all hamsters was normal. Cholesterol gallstones were found in 2 hamsters at the end of the 4th week. The geometric center values for the experimental and control groups were 2.3891 +/- 0.3923 vs. 2.7730 +/- 0.5283, at the end of week 3; 1.8148 +/- 0.4312 vs. 3.2294 +/- 1.1613 at week 4; 1.8451 +/- 0.3700 vs. 2.9075 +/- 0.3756 at week 5; and 1.8025 +/- 0.3413 vs. 3.0920 +/- 0.5622 at week 6. CONCLUSION: A high cholesterol diet can significantly reduce the intestinal transit function and facilitate the formation of cholesterol gallstones.

Overexpression of sterol carrier protein-2 mRNA in patients with cholesterol gallstones

BACKGROUND: Hypersecretion of biliary cholesterol is believed to be one of the important causes of lithogenic bile. Sterol carrier protein-2 ( SCP2 ) participates in choles- terol trafficking and metabolism and may play a key role in cholesterol gallstone formation. This study was undertaken to investigate the expression of liver SCP2 mRNA in pa- tients with cholesterol gallstone and those patients with non-cholesterol gallstone. METHODS: The expression of liver SCP2 mRNA was studi- ed in 36 patients with cholesterol gallstone and 30 patients with non-cholesterol gallstone by reverse transcription-poly- merase chain reaction (RT-PCR). RESULT: The expression of SCP2 mRNA was increased more significantly in patients with cholesterol gallstone than in patients with non-cholesterol gallstone. CONCLUSION: The SCP2 gene was overexpressed in pa- tients with cholesterol gallstone, indicating that SCP2 may be one of the important causes of cholesterol gallstone.

Albiflorin Granule significantly decreased the cholesterol gallstone formation by the regulation of insulin transduction signal

Objective:To study the mechanism of insulin resistance in the cholesterol gallstone formation from insulin signal transduction pathway so as to reveal the possible mechanism and the effective role of Albiflorin Granule on preventing the cholesterol gallstones.Methods:Serum triglycerides(TG),free fatty acid(FFA),and total cholesterol(TC) from different groups were measured and liver cells Ins R,PKB,IKK-β protein expression levels were detected by western blotting.Results:Albiflorin significantly decreased the cholesterol gallstone formation rate,increased glucose infusion rate in gallstone guinea pigs and improved insulin resistance.Compared with the normal group,insulin receptor and PKB protein expression in GS group were significantly reduced.IKK-β protein in the GS group increased significantly and Albiflorin could reduce IKK-β protein expression in guinea pig liver cells.Conclusions:The model of insulin resistance in cholesterol gallstone guinea pig was successfully established,which plays an important role in the cholesterol gallstone formation.All aspects of insulin signaling pathway are involved in gallstone formation.Albiflorin can regulate various aspects of insulin signal transduction pathway to prevent the formation of gallbladder.

A comparative study of changing patterns of concanavalin A-binding proteins in early stage of cholesterol gallstone

IM To elucidate the importance and the changing patterns of biliary concanavalin Abinding proteins (CPs) in the early stage of cholesterol gallstone.METHODS CPs concentrations and nucleation activities were measured by lectin affinity chromatography in biles of patients with cholesterol gallstone, pigment gallstone, gallbladder cholesterosis and nonbiliary diseases.RESULTS The concentrations of CPs were much higher in patients with cholesterol gallstone (039g/L±011g/L, n=36, P<001) or gallbladder cholesterosis (040g/L±009g/L, n=9, P<001) than in those with pigment gallstone (026g/L±012g/L, n=7) and/or nonbiliary diseases (027g/L±009g/L, n=10). Pronucleating activities were much stronger in patients with cholesterol gallstones (nucleation time ratio: 057±021, n=5, P<001 vs pigment gallstone and/or nonbiliary diseases) and gallbladder cholesterosis (nucleation time ratio: 044±023, n=5, P<001 vs pigment gallstone or nonbilliary diseases). The binding percentages of CPs to model biliary vesicles were also higher in patients with cholesterol gallstones (n=6) than those with pigment gallstones (n=6) (24%±09% vs 09%±05%, P<001).CONCLUSION Hypersecretion of CPs, especially those in vesicular phase may be the important changes in the early stage of cholesterol gallstone.

High level of deoxycholic acid in human bile does not promote cholesterol gallstone formation

AIM:To study whether patients with excess deoxycholic acid (DCA) differ from those with normal percentage of DCA with respect to biliary lipid composition and cholesterol saturation of gallbladder bile. METHODS:Bile was collected during operation through puncturing into the gallbladder from 122 cholesterol gallstone patients and 46 gallstone-free subjects undergoing cholecystectomy.Clinical data,biliary lipids,bile add composition, presence of crystals and nucleation time were analyzed. RESULTS:A subgroup of gallstone patients displayed a higher proportion of DCA in bile than gallstone free subjects. By choosing a cut-off level of the 90th percentile,a group of 13 gallstone patients with high DCA levels (mean 50 percent of total bile acids) and a large group of 109 patients with normal DCA levels (mean 21 percent of total bile acids) were obtained.The mean age of the patients with high DCA levels was higher than that of the group with normal levels (mean age:62 years vs45 years) and so was the mean BMI (28.3 vs.24.7).Plasma levels of cholesterol and triglycerides were slightly higher in the DCA excess groups compared with those in the normal DCA group.There was no difference in biliary lipid composition,cholesterol saturation,nucleation time or occurrence of cholesterol crystals in bile between patients with high and normal levels of DCA. CONCLUSION:Gallstone patients with excess DCA were of older age and had higher BMI than patients with normal DCA.The two groups of patients did not differ with respect to biliary lipid composition,cholesterol saturation,nucleation time or occurrence of cholesterol crystals.It is concluded that DCA in bile does not seem to contribute to gallstone formation in cholesterol gallstone patients.

Effects of long term hydrophilic bile acid therapy on in vitro contraction of gallbladder muscle strips in patients with cholesterol gallstones

AIM:To evaluate ursodeoxycholic acid (UDCA) therapy on the in vitro contraction of gallbladder smooth muscle strips from cholesterol gallstone patients. METHODS:The contraction forces of gallbladder smooth muscle strips from 28 patients with cholesterol gallstones treated with UDCA were compared with contraction forces from 14 untreated patients. The strips were stimulated with increasing concentrations of cholecystokinin-8 (CCK-8). RESULTS:Although the contraction forces that developed in response to CCK-8 were higher in strips from specimens of UDCA treated patients compared to untreated patients,longer treatment periods (6-wk) caused more contraction responses than the short treatment period of 3-wk (F = 19.297,1.85 ± 0.22 g vs 1.70 ± 0.10 g,P < 0.01). Contraction forces developed with maximal stimulation with KCl in the 6-wk treatment group were also higher than contraction forces in the untreated group (F = 4.274,3.77 ± 0.45 g vs 3.30 ± 0.30 g,P < 0.05). CONCLUSION:Six-week UDCA treatment caused an increase in contractions of muscle strips from patients with cholesterol gallstones when compared to shorter treatment administration or controls. We suggest that extending UDCA treatment periods may cause more effective contractions in the gallbladder,and thereby increase the rate of response to treatment.

Research progress on the immune microenvironment of the gallbladder in patients with cholesterol gallstones

Cholesterol gallstones are very common in hepatobiliary surgery and have been studied to a certain extent by doctors worldwide for decades.However,the mechanism of cholesterol gallstone formation is not fully understood,so there is currently no completely effective drug for the treatment and prevention of cholesterol gallstones.The formation and development of cholesterol gallstones are caused by a variety of genetic and environmental factors,among which genetic susceptibility,intestinal microflora disorders,impaired gallbladder motility,and immune disorders are important in the pathogenesis of cholesterol gallstones.This review focuses on recent advances in these mechanisms.We also discuss some new targets that may be effective in the treatment and prevention of cholesterol gallstones,which may be hot areas in the future.

Cholesterol gallstones:Focusing on the role of interstitial Cajal-like cells

with a complex and multifactorial etiology.Declined gallbladder motility reportedly contributes to CG pathogenesis.Furthermore,interstitial Cajal-like cells(ICLCs)are reportedly present in human and guinea pig gallbladder tissue.ICLCs potentially contribute to the regulation of gallbladder motility,and aberrant conditions involving the loss of ICLCs and/or a reduction in its pacing potential and reactivity to cholecystokinin may promote CG pathogenesis.This review discusses the association between ICLCs and CG pathogenesis and provides a basis for further studies on the functions of ICLCs and the etiologies of CG.

Piperine prevents cholesterol gallstones formation in mice

Biliary cholesterol may contribute to the formation of cholesterol gallstones, regulation of these levels could be a useful therapeutic strategy for gallstones disease. Piperine （PA） is a potential cholesterol lowering a- gent. In this study, we assessed the effect and mechanism of PA in preventing cholesterol gallstones formation in- duced by feeding lithogenic diet containing high cholesterol levels to mice. C57BL/6 inbred mice were fed lithogen- ic or chow diets for 10 weeks, with or without PA （ 15, 30 and 60 mg ~ kg-1 ） or ursodeoxycholic acid （ UDCA, 60 mg ~ kg-1） administration. Cholesterol, phospholipids and crystals in bile, the lipid in serum, pathological chan- ges and proteins expression in liver were analyzed. The results showed that PA could decrease the cholesterol poten- cy and crystals in bile, reduce total cholesterol （TC）, triglycerides （TG） and increase high-density lipoprotein / low-density lipoprotein （HDL/LDL） levels in serum. Furthermore, PA treatment reduced liver lipid peroxidation and protected hepatobiliary cells from liver injury by decreasing malondialdehyde （MDA） and increasing superoxide dismutase （SOD）. In addition, PA inhibited the expression of ATP-binding cassette transporters G5/8 （ABCG5/ 8 ） and liver X receptor （LXR） in liver, and reduced cholesterol transport from the hepatocytes to the gallbladder. It may be the mechanism of PA in preventing cholesterol gallstones formation. PA as a potential drug for prevention cholesterol gallstones merits further investigation.

Bile acids in serum and bile of patients with cholesterol gallstone

IM To analyze serum bile acids and biliary lipids of patients with cholesterol gallstone(CS) and explore the relationship between deoxycholic acid (DCA) and CS disease.METHODS Analysis of bile acids in serum was done with gaschromatography in two groups: CS group (n=151) and control group (n=256). Serum bile acids and biliary lipids were also studied in 90 matched samples..RESULTS The serum DCA was 0955μmol/L±0078μmol/L in CS group, which was more than that of control group (0696μmol/L±0047μmol/L), P<001. The ratio of DCA/chenodeoxycholic acids (CDCA) was 176±030 in CS group, about two times that in control group (092±014). The mole percent of DCA in bile was positively related to cholesterol saturation index (CSI) (P<001) and the mole percent of CDCA in bile negatively to CSI (P=001). There was correlation between the mole percent of DCA, CDCA and cholic acid in bile and in serum.CONCLUSION It is suggested that DCA is lithogenic and the increased amount of DCA or the ratio of DCA/CDCA in serum may be one of the features of cholesterol gallstone patients.

建立HPLC-CS检测腮腺液胆固醇的方法及应用

目的 对腮腺液胆固醇检测方法进行探索和条件优化,探讨其与龋病发生的相关性.方法 采用Slominary方法收集不同龋敏感者的腮腺液,冷冻干燥后用Folch方法提取脂质.采用Alltech ODS作为预富集柱,采用反向HPLC 对腮腺液胆固醇进行定性、定量检测.结果 预富集流动相：乙腈/（甲醇/0.1 g/dl磷酸水溶液=6∶1）=5/95（V/V）,流速0.3 ml/min;分析流动相：乙腈/甲醇=1/9 （V/V）,流速1.0 ml/min,CS-HPLC优于HPLC,t=5.473,P〈0.05;高龋组腮腺液胆固醇含量明显高于无龋组.结论 CS-HPLC可以有效实现唾液痕量物质的预富集和净化,提高检测的灵敏度;腮腺液胆固醇与龋病具有相关性.

Gallstones in patients with liver cirrhosis:Incidence,etiology,clinical and therapeutical aspects

Gallstones occur in about one third of the patients having liver cirrhosis.Pigment gallstones are the most frequent type,while cholesterol stones represent about15%of all stones in cirrhotics.Increased secretion of unconjugated bilirubin,increased hydrolysis of conjugated bilirubin in the bile,reduced secretion of bile acids and phospholipds in bile favor pigment lithogenesis in cirrhotics.Gallbladder hypomotility also contributes to lithogenesis.The most recent data regarding risk factors for gallstones are presented.Gallstone prevalence increases with age,with a ratio male/female higher than in the general population.Chronic alcoholism,viral C cirrhosis,and non-alcoholic fatty liver disease are the underlying liver diseases most often associated with gallstones.Gallstones are often asymptomatic,and discovered incidentally.If asymptomatic,expectant management is recommended,as for asymptomatic gallstones in the general population.However,a closer follow-up of these patients is necessary in order to earlier treat symptoms or complications.For symptomatic stones,laparoscopic cholecystectomy has become the therapy of choice.Child-Pugh class and MELD score are the best predictors of outcome after cholecystectomy.Patients with severe liver disease are at highest surgical risk,therefore gallstone complications should be treated using noninvasive or minimally invasive procedures,until stabilization of the patient condition.

Biliary cholesterol secretion: More than a simple ABC

Biliary cholesterol secretion is a process important for 2 major disease complexes, atherosclerotic cardiovascular disease and cholesterol gallstone disease. With respect to cardiovascular disease, biliary cholesterol secretion is regarded as the f inal step for the elimination of cholesterol originating from cholesterol-laden macrophage foam cells in the vessel wall in a pathway named reverse cholesterol transport. On the other hand, cholesterol hypersecretion into the bile is considered the main pathophysiological determinant of cholesterol gallstone formation. This review summarizes current knowledge on the origins of cholesterol secreted into the bile as well as the relevant processes and transporters involved. Next to the established ATP-binding cassette (ABC) transporters mediating the biliary secretion of bile acids (ABCB11), phospholipids (ABCB4) and cholesterol (ABCG5/G8), special attention is given to emerging proteins that modulate or mediate biliary cholesterol secretion. In this regard, the potential impact of the phosphatidylserine flippase ATPase class Ⅰ type 8B member 1, the Niemann Pick C1-like protein 1 that mediatescholesterol absorption and the high density lipoprotein cholesterol uptake receptor, scavenger receptor class B type Ⅰ, is discussed.

Serum leptin levels and insulin resistance are associated with gallstone disease in overweight subjects

AIM: To establish an association between the serum leptin levels and the development of gallstone disease (GD).METHODS: We carried out a non-matched case-controlled study in a university hospital in Mexico City. Two hundred and eighty-seven subjects were included: 97 cases with gallstones and 190 controls. Body mass index (BMI), fasting plasma leptin, insulin, serum lipid, and lipoprotein levels were measured. Insulin resistance was calculated by homeostasis model assessment (HOMA-IR). Unconditional logistic regression analysis (univariate and multivariate)stratified by BMI was used to calculate the risk of GD.RESULTS: The multivariate conditional regression analysis revealed a model for those patients with BMI ＜30. The selected variables in the model were HOMA-IR index with OR = 1.31, P= 0.02 and leptin higher than median with OR = 2.11, P= 0.05. In the stratum of BMI ≥30, we did not find a useful model.CONCLUSION: We concluded that insulin resistance and the development of GD appears to be associated with serum leptin levels in subjects with overweight, but not in obese subjects with similar metabolic profiles.

Sex-specific and Dose-response Relationship between the Incidence of Gallstones and Components of the Metabolic Syndrome in Jinchang Cohort:A Prospective Study

Gallstones is a very common gastrointestinal disease,involving the formation of masses in the gall bladder or biliary tract,due to abnormally high concentrations of cholesterol or bilirubin in the bile[1].There is a 10%–20%prevalence of gallstones worldwide,and>20%of patients with gallstones show clinical symptoms,implying a substantial disease burden[1].

Extensive Quantitative Analysis of Gallstones

Background: The chemical composition of gallstones is essential to study aetiopathogenesis of gallstone disease. Objective: To determine the composition of gallstones in a Sudanese population. Patients and methods: It describes an extensive quantitative analysis of gallstones from patients presented with symptomatic gall stone disease and treated by cholecystectomy after the acceptance of the pre-given informed consent in Khartoum teaching hospital in the period between Jan 2010 and Dec 2010. Using a pretested questionnaire data collected from and analyzed statistically by SPSS computer program version 21. Results: Data are analyzed from 94 patients (six males and 88 females). Cholesterol stones showed a significantly higher cholesterol content than pigment stones (p = 0.0042), though not significantly higher than mixed stones. Their phospholipids content and inorganic phosphates were higher than in the other types of stones and oxalate content was significantly elevated in comparison with mixed stones (p = 0.0402). In mixed stones, the cholesterol, bile acids, and bilirubin were intermediate between cholesterol and pigment stones, whereas triglycerides were significantly more than pigment stones (p = 0.0007). Bilirubin (p = 0.0021) and bile acids (p = 0.0016) were significantly higher than cholesterol stones (p = 0.0001) and (p = 0.0001) respectively. However, they contained the lowest amounts of sodium, potassium, magnesium, and oxalate. In pigment stones, bilirubin was significantly higher (p = 0.0001) than both groups. Conclusion: Collaborations between surgeons, nutritionists, biochemists, and physicians should be stimulated in future studies to define the different types of gall stones in different areas in Sudan and the relevancy of such types with diets’ traditions.