Effect of peroxisome proliferator-activated receptor-gamma ligand on inflammation of human gallbladder epithelial cells

AIM: To investigate the effect of peroxisome proliferatoractivated receptor gamma (PPAR-γ) and its ligand, ciglitazone, on inflammatory regulation of human gallbladder epithelial cells (HGBECs) and to assess the effect of human epithelial growth factor (hEGF) on growth of HGBECs. METHODS: HGBECs were cultured in media containing hEGF or in hEGF-free media. HGBECs were divided into normal control group, inflammatory control group and ciglitazone group (test group). Inflammatory control group and ciglitazone group were treated with 5 μg/L of human interleukin-1β (hIL-1β) to make inflammatory model of HGBECs. The ciglitazone group was treated with various concentrations of ciglitazone, a potent ligand of PPAR-γ. Subsequently, interleukin-8 (IL-8), IL-6, and tumor necrosis factor-α (TNF-α) concentrations in all groups were measured. The data were analyzed statistically. RESULTS: HGBECs were cultured in medium successfully. The longevity of HGBECs in groups containing hEGF was longer than that in hEGF-free groups. So was the number of HGBECs. The longest survival time of HGBEC was 25 d. The inflammatory model of HGBECs was obtained by treating with hIL-lp. The concentrations of IL-6 and IL-8 in ciglitazone group were lower than those in inflammatory control group (P<0.05). The secretion of IL-6 in inflammatory control group was higher (350.31±37.05 μg/L) than that in normal control group (50.0±0.00 μg/L, P<0.001). Compared to normal control group, IL-8 concentration in inflammatory control was higher (P<0.05). CONCLUSION: hEGF improves the growth of HGBECs in vitro. Ciglitazone inhibits the inflammation of HGBECs in vitro and has potential therapeutic effect on cholecystitis in vivo.     

Effect of three nonpeptide cholecystokinin antagonists on human isolated gallbladder

Cholecystokinin is the most important stimulant of postprandial gallbladder contraction, and a regulator of gallbladder fasting tone. The aim of this study was to evaluate the effect of dexloxiglumide on isolated human gallbladder contraction induced by cholecystokinin–octapeptide and to compare this effect to that of lorglumide and amiglumide, two glutaramic acid analogs of dexloxiglumide. The negative logarithms of the antagonist dissociation constant (pK_B) values were 7.00 ± 0.14, 6.95 ± 0.11, and 6.71 ± 0.10 for lorglumide, dexloxiglumide, and amiglumide, respectively. Dexloxiglumide produced a concentration-dependent rightward shift of the cholecystokinin–octapeptide curve, without affecting its maximal response. A similar effect was obtained both with lorglumide and amiglumide. Moreover, the slopes for the three antagonists did not differ significantly from unity. These data show that the three molecules have a potent antagonistic effect, of a competitive nature, on gallbladder cholecystokinin type 1 receptors. It may be concluded that dexloxiglumide, lorglumide, and amiglumide exhibit a promising therapeutic profile for biliary colic and other gastrointestinal disorders in which CCK₁ receptors play important physiological roles.     

Effect of gastrin-releasing peptide (GRP) on guinea pig gallbladder contrction in vitro

Few studies have reported the effects of gastrin-releasing peptide (GRP)/bombesin on the guineas pig gallbladder, and the results are contradictory. Because such contradictory results may, in part, be due to technical factors, we investigated the effect of GRP on guinea pig gallbaladder smooth muscle, using a improved horizontal organ bath. The guinea pigs were killed and the gallbladder was removed. Four longitudinal uscle strips (2×12mm) were suspended in Krebs-Ringer solution at 37°C and aerated with 95% O₂ and 5% CO₂. The mechanical activity of the strips was recorded isotonically by displacement-voltage transducers. via L-arms, to which a piezoelectric element with a frequency of 100Hz and movement of 50μm was applied. GRP contracted gallbladder muscle strips dose dependently, but the calculated maximal response was 22.4% and 20.1% of the acetylcholine-and cholecystokinin octapeptide (CCK8)-induced responses, respectively. The GRP-induced contraction was unaffected by the muscarinic blocker, atropine, or by the CCK receptor antagonist, loxiglumide. It is concluded that GRP weakly, but apparently directly, stimulates guinea pig gallbladder contraction.     

Undifferentiated spindle-cell carcinoma of the gallbladder: an immunohistochemical study

A case of undifferentiated spindle-cell carcinoma of the gallbladder is described. A 72-year-old man presented with right hypochondralgia and fever. Imaging studies revealed a well-demarcated solid tumor (with a necrotic center) in the gallbladder that invaded the liver and transverse colon. On gross examination of the surgical specimen, the cut surface of the polypoid tumor showed nodular invasive growth. Microscopically, the tumor was composed of atypical spindle-shaped tumor cells that proliferated in a whirling or interlacing pattern. The tumor also showed foci with a malignant epithelial component that simulated a carcinosarcoma. Immunohistochemically, the biphasic differentiation of the tumor was highlighted by the different immunoreactivity to antibodies against cytokeratins, epithelial membrane antigen (EMA), and vimentin shown by the malignant epithelial components and the spindle-cell components. However the latter showed faint positivity for cytokeratin antibody. These results suggested that the spindle-cell carcinoma of the gallbladder originated from cholecystic mucosa and showed sarcomatous reaction or dedifferentiation, as indicated by the presence of vimentin-positive cells. The proliferation index, as detected by ki-67, in the spindle-cell component was higher than that in the epithelial component, which may account for the more aggressive biological behavior of the spindle-cell component.     

Successful treatment of gallbladder carcinoma producing alpha-fetoprotein with segmental adenomyomatosis

The successful treatment of hepatoid adenocarcinoma of the gallbladder with elevated serum alpha-fetoprotein (1243 ng/ml) and segmental adenomyomatosis in a 58-year-old woman is described. The woman had alpha-fetoprotein (AFP)-producing carcinoma of the gallbladder with regional lymph node metastasis and was treated by extended radical resection and postoperative adjuvant chemotherapy. She is alive, showing normal serum AFP concentration and no recurrence, 57 months after surgery. The tumor cells were stained immunohistochemically for AFP by the peroxidase anti-peroxidase method. Serum AFP reactivity to concanavalin A and lentil lectin was similar to the pattern shown in hepatocellular carcinoma. Only a few cases of AFP-producing gallbladder carcinoma have been reported and there have been no reports of long-term survivors. The combination of aggressive radical resection and chemotherapy seems to have been effective for achieving long-term survival without liver metastasis.     

Segmental adenomyomatosis of the gallbladder predisposes to cholecystolithiasis

Background/Purpose

The aim of the present study was to clarify the association between adenomyomatosis of the gallbladder and cholecystolithiasis.

Methods

A cholecystectomy was performed for cholelithiasis or various other conditions in 1099 patients, of whom 608 had cholecystolithiasis. Adenomyomatosis of the gallbladder was classified as one of three variants: segmental, fundal, and diffuse. Segmental adenomyomatosis has an annular stricture dividing the gallbladder lumen into the “neck compartment” and the “fundal compartment”. Bile lipid analysis was performed in 8 patients with segmental adenomyomatosis.

Results

Adenomyomatosis of the gallbladder was observed in 156 patients (14.2%), of whom 99 had segmental adenomyomatosis, 54 had fundal adenomyomatosis, and 3 had diffuse adenomyomatosis. The prevalence of cholecystolithiasis was higher in patients with segmental adenomyomatosis (88.9%) than in those without adenomyomatosis (52.3%; P < 0.001). Gallstones were detected earlier in patients with segmental adenomyomatosis than in those without (P < 0.001) and were located predominantly in the fundal compartment. Bile in the fundal compartment had lower concentrations of total bile acids (P = 0.012), with an increased cholesterol saturation index (P = 0.012), compared to bile in the neck compartment.

Conclusions

Segmental adenomyomatosis is a condition predisposing to cholecystolithiasis, probably due to the lithogenic environment in the fundal compartment. Fundal or diffuse adenomyomatosis appears to be unrelated to cholecystolithiasis.

     

Prostaglandin E2 stimulates ion transport in prairie dog gallbladder

The effects of prostaglandins, and specifically prostaglandin E₂, on gallbladder ion transport were examined in the prairie dog. Gallbladders were mounted in an Ussing chamber and baseline short-circuit current, potential difference, and tissue resistance were measured. Addition of arachidonic acid (10^–4 M, mucosal surface) produced sustained elevations in short-circuit current and potential difference (P<0.05), with mild reductions in resistance. In a second set of tissues, indomethacin exposure (10^–6 M) resulted in a significant (P<0.02) decrease in short-circuit current and potential difference, with an increase in resistance. Subsequent addition of prostaglandin E₂ (10^–7 M, serosal surface) fully reversed these changes and led to a significant increase in short-circuit current and potential difference (P<0.001) with a return of resistance to baseline values. These findings suggest that endogenous prostaglandins mediate gallbladder ion transport.Financial support received from the Veterans Administration. Dr. Saunders-Kirkwood was supported by the Claude E. Welch Research Fellowship, Department of Surgery, Massachusetts General Hospital.     

Haplotype analysis of signal peptide (insertion/deletion) and XbaI polymorphisms of the APOB gene in gallbladder cancer.

PURPOSE: The incidence of gallbladder cancer (GBC) is usually paralleled by the prevalence of gallstone disease, and genes of cholesterol metabolism have been implicated in gallstone disease. The XbaI and insertion/deletion (ins/del) polymorphism of Apolipoprotein B (APOB) appears to influence cholesterol homoeostasis and possibly risk for gallstone disease. We examined the effect of these polymorphisms individually as well as their haplotypes on GBC and gallstone patients in North Indian population. METHODS: The study comprises 123 consecutive cases of proven GBC, 172 cases of gallstone and 232 healthy subjects of similar age and sex. The genomic DNA was extracted from peripheral blood leucocytes and genotyping was performed using polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism. RESULTS: In a case-control study, APOB XbaI and ins/del polymorphisms were not significantly associated with risk of GBC. Using the expectation maximization algorithm, four haplotypes were obtained, and haplotype X(+),D was found to be significantly higher in GBC patients without stone in comparison with healthy subjects [odds ratio (OR) 2.9, 95% confidence interval 1.2-6.6 P=0.012]. CONCLUSIONS: The X(+),D haplotype of APOB is associated with increased risk for development of GBC and the risk is not modified in the presence of gallstones.     

Primary sclerosing cholangitis in which differential diagnosis from gallbladder carcinoma was difficult

We report a case of localized primary sclerosing cholangitis (PSC) which was difficult to distinguish from gallbladder carcinoma. A 75-year-old woman with elevated serum bilirubin was hospitalized and underwent endoscopic nasobiliary drainage (ENBD). There was no history of diseases such as gallbladder stone, pancreatitis, or ulcerative colitis. Cholangiography through the ENBD tube showed localized stenosis of the common bile duct; the gallbladder could not be seen. Angiography showed no encasement of the hepatic artery. Ultrasonography showed a tumor in the cystic duct, and the tumor had invaded the gallbladder and common bile duct. We diagnosed gallbladder carcinoma on radioimaging, and performed an S4aS5 subsegmentectomy of the liver and resection of the extrahepatic biliary tree. Pathologically, no malignant cells were detected, and fibrosis around bile ducts and infiltration of inflammatory cells into hepatic tissue were found. It is well known that PSC is sometimes difficult to differentially diagnose from cholangiocarcinoma. Our case is of high interest because ultrasonography showed findings suggestive of gallbladder carcinoma. It is therefore necessary to keep the possibility of PSC in mind for the diagnosis and treatment of such localized biliary stenosis.     

Emergency Biliary Sonography: Utility of Common Bile Duct Measurement in the Diagnosis of Cholecystitis and Choledocholithiasis

Background

Measurement of the common bile duct (CBD) has traditionally been considered an integral part of gallbladder sonography, but accurate identification of the CBD can be difficult for novice sonographers.

Objective

To determine the prevalence of isolated sonographic CBD dilation in emergency department (ED) patients with cholecystitis or choledocholithiasis without laboratory abnormalities or other pathologic findings on biliary ultrasound.

Methods

We conducted a retrospective chart review on two separate ED patient cohorts between June 2000 and June 2010. The first cohort comprised all ED patients undergoing a biliary ultrasound and subsequent cholecystectomy for presumed cholecystitis. The second cohort consisted of all ED patients receiving a biliary ultrasound who were ultimately diagnosed with choledocholithiasis. Ultrasound data and contemporaneous laboratory values were collected. Postoperative gallbladder pathology reports and endoscopic retrograde cholangiopancreatography (ERCP) reports were used as the criterion standard for final diagnosis.

Results

Of 666 cases of cholecystitis, there were 251 (37.7%) with a dilated CBD > 6 mm and only 2 cases (0.3%; 95% confidence interval [CI] 0.0–0.7%) of isolated CBD dilation with an otherwise negative ultrasound and normal laboratory values. Of 111 cases of choledocholithiasis, there were 80 (72.0%) with a dilated CBD and only 1 case (0.9%; 95% CI 0.0–2.7%) with an otherwise negative ultrasound and normal laboratory values.

Conclusion

The prevalence of isolated sonographic CBD dilation in cholecystitis and choledocholithiasis is <1%. Omission of CBD measurement is unlikely to result in missed cholecystitis or choledocholithiasis in the setting of a routine ED evaluation with an otherwise normal ultrasound and normal laboratory values.