Increased renal expression of bilirubin glucuronide transporters in a rat model of obstructive jaundice

2002 ◽  
Vol 282 (4) ◽  
pp. G656-G662 ◽  
Author(s):  
Yuji Tanaka ◽  
Yoshinao Kobayashi ◽  
Esteban C. Gabazza ◽  
Kunihiro Higuchi ◽  
Toshinori Kamisako ◽  
...  
Keyword(s):  
Bile Acid ◽  
Obstructive Jaundice ◽  
Protein Expression ◽  
Mrna Expression ◽  
Sham Operation ◽  
Compensatory Mechanism ◽  
Human Bile ◽  
Duct Ligation ◽  
Pah Clearance ◽  
Conjugated Bilirubin

Regulation of bilirubin glucuronide transporters during hyperbilirubinemia in hepatic and extrahepatic tissues is not completely clear. In the present study, we evaluated the regulation of the bilirubin glucuronide transporters, multidrug resistance-associated proteins (MRP)2 and 3, in rats with obstructive jaundice. Bile duct ligation (BDL) or sham operation was performed in Wistar rats. Liver and kidneys were removed 1, 3, and 5 days after BDL ( n = 4, in each group). Serum and urine were collected to measure bilirubin levels just before animal killing. MRP2 And MRP3 mRNA expressions were determined by real-time RT-PCR. Protein expression of MRP2 and MRP3 was determined by Western blotting. Renal MRP2 function was evaluated by para-aminohippurate (PAH) clearance. The effect of conjugated bilirubin, unconjugated bilirubin, human bile, and sulfate-conjugated bile acid on MRP2 gene expression was also evaluated in renal and hepatocyte cell lines. Serum bilirubin and urinary bilirubin excretion increased significantly after BDL. In the liver, the mRNA expression of MRP2 decreased 59, 86, and 82%, and its protein expression decreased 25, 74, and 93% compared with sham-operated animals after 24, 72, and 120 h of BDL, respectively. In contrast, the liver expression of MRP3 mRNA increased 138, 2,137, and 3,295%, and its protein expression increased 560, 634, and 612% compared with sham-operated animals after 24, 72, and 120 h of BDL, respectively. On the other hand, in the kidneys, the mRNA expression of MRP2 increased 162, 73, and 21%, and its protein expression increased 387, 558, and 472% compared with sham-operated animals after 24, 72, and 120 h of BDL, respectively. PAH clearance was significantly increased after BDL. The mRNA expression of MRP2 increased in renal proximal tubular epithelial cells after treatment with conjugated bilirubin, sulfate-conjugated bile acid or human bile. Upregulation of MRP2 in the kidneys and MRP3 in the liver may be a compensatory mechanism to improve bilirubin clearance during obstructive jaundice.

Ammonia reduction with ornithine phenylacetate restores brain eNOS activity via the DDAH-ADMA pathway in bile duct-ligated cirrhotic rats

2012 ◽  
Vol 302 (1) ◽  
pp. G145-G152 ◽  
Author(s):  
Vairappan Balasubramaniyan ◽  
Gavin Wright ◽  
Vikram Sharma ◽  
Nathan A. Davies ◽  
Yalda Sharifi ◽  
...  
Keyword(s):  
Bile Duct ◽  
Protein Expression ◽  
Ammonia Concentration ◽  
Sham Operation ◽  
Sprague Dawley ◽  
Sprague Dawley Rats ◽  
Duct Ligation ◽  
Tnf Α ◽  
No Signaling ◽  
Brain Water

Ammonia is central in the pathogenesis of hepatic encephalopathy, which is associated with dysfunction of the nitric oxide (NO) signaling pathway. Ornithine phenylacetate (OP) reduces hyperammonemia and brain water in cirrhotic animals. This study aimed to determine whether endothelial NO synthase activity is altered in the brain of cirrhotic animals, whether this is associated with changes in the endogenous inhibitor, asymmetric-dimethylarginine (ADMA) and its regulating enzyme, dimethylarginine-dimethylaminohydrolase (DDAH-1), and whether these abnormalities are restored by ammonia reduction using OP. Sprague-Dawley rats were studied 4-wk after bile duct ligation (BDL) ( n = 16) or sham operation ( n = 8) and treated with placebo or OP (0.6 g/kg). Arterial ammonia, brain water, TNF-α, plasma, and brain ADMA were measured using standard techniques. NOS activity was measured radiometrically, and protein expression for NOS enzymes, ADMA, DDAH-1, 4-hydroxynonenol (4HNE), and NADPH oxidase (NOX)-1 were measured by Western blotting. BDL significantly increased arterial ammonia ( P < 0.0001), brain water ( P < 0.05), and brain TNF-α ( P < 0.01). These were reduced significantly by OP treatment. The estimated eNOS component of constitutive NOS activity was significantly lower ( P < 0.05) in BDL rat, and this was significantly attenuated in OP-treated animals. Brain ADMA levels were significantly higher and brain DDAH-1 significantly lower in BDL compared with sham ( P < 0.01) and restored toward normal following treatment with OP. Brain 4HNE and NOX-1 protein expression were significantly increased in BDL rat brain, which were significantly decreased following OP administration. We show a marked abnormality of NO regulation in cirrhotic rat brains, which can be restored by reduction in ammonia concentration using OP.

scholarly journals The Effect of Biliary Decompression on Bacterial Translocation in Jaundiced Rats

HPB Surgery ◽  
1993 ◽  
Vol 7 (2) ◽  
pp. 99-110 ◽  
Author(s):  
Jin Wen Ding ◽  
Roland Andersson ◽  
Vasile Soltesz ◽  
Roger Willén ◽  
Steffen Loft ◽  
...  
Keyword(s):  
Liver Function ◽  
Obstructive Jaundice ◽  
Bacterial Translocation ◽  
Serum Alkaline Phosphatase ◽  
Morphological Changes ◽  
Sham Operation ◽  
Biliary Decompression ◽  
Septic Complications ◽  
Sham Operation Group ◽  
Duct Ligation

Patients with obstructive jaundice are prone to septic complications after biliary tract operations. Restoring bile flow to the intestine may help to decrease the complication rate. The present study is aimed at evaluating the effect of biliary decompression on bacterial translocation in jaundiced rats.Sixty-six male Sprague-Dawley rats were randomly allocated to six groups subjected to common bile duct ligation (CBDL) and transection (groups 2–6) or sham operation (group 1). In groups and 2 the incidence of enteric bacterial translocation was determined 2 weeks after sham operation or CBDL. In groups 3–6, biliary decompression was achieved by performing a choledochoduodenostomy after 2 weeks of biliary decompression. Bacterial translocation was then studied 1,2,3 and 5 weeks following biliary decompression.The rate of bacterial translocation to mesenteric lymph nodes in obstructive jaundice was significantly higher as compared with controls, and decreased with time to nil three weeks following biliary decompression. The incidence of bacterial translocation was closely correlated (r = 0.844; p = 0.034) with serum alkaline phosphatase activity and seemed to fit with the morphological changes noted in the small intestine. The decrease in bacterial translocation, however, lags behind the recovery of liver function as measured by routine liver function tests and antipyrine clearance.Obstructive jaundice thus promotes bacterial translocation in the rat. Biliary decompression gradually decreases the rate of bacterial translocation.

scholarly journals Increased cholesterol 7α-hydroxylase expression and size of the bile acid pool in the lactating rat

2008 ◽  
Vol 294 (4) ◽  
pp. G1009-G1016 ◽  
Author(s):  
Clavia Ruth Wooton-Kee ◽  
David E. Cohen ◽  
Mary Vore
Keyword(s):  
Bile Acid ◽  
Protein Expression ◽  
Mrna Expression ◽  
Bile Acids ◽  
Fold Increase ◽  
Bile Acid Pool ◽  
Hydrophobicity Index ◽  
Acid Pool ◽  
Bile Acid Transporters ◽  
Cholic Acids

Maximal bile acid secretory rates and expression of bile acid transporters in liver and ileum are increased in lactation, possibly to facilitate increased enterohepatic recirculation of bile acids. We determined changes in the size and composition of the bile acid pool and key enzymes of the bile acid synthetic pathway [cholesterol 7α-hydroxylase (Cyp7a1), sterol 27-hydroxylase (Cyp27a1), and sterol 12α-hydroxylase (Cyp8b1)] in lactating rats relative to female virgin controls. The bile acid pool increased 1.9 to 2.5-fold [postpartum (PP) days 10, 14, and 19–23], compared with controls. A 1.5-fold increase in cholic acids and a 14 to 20% decrease in muricholic acids in lactation significantly increased the hydrophobicity index. In contrast, the hepatic concentration of bile acids and small heterodimer partner mRNA were unchanged in lactation. A 2.8-fold increase in Cyp7a1 mRNA expression at 16 h (10 h of light) demonstrated a shift in the diurnal rhythm at day 10 PP; Cyp7a1 protein expression and cholesterol 7α-hydroxylase activity were significantly increased at this time and remained elevated at day 14 PP but decreased to control levels by day 21 PP. There was an overall decrease in Cyp27a1 mRNA expression and a 20% decrease in Cyp27a1 protein expression, but there was no change in Cyp8b1 mRNA or protein expression at day 10 PP. The increase in Cyp7a1 expression PP provides a mechanism for the increase in the bile acid pool.

Abstract P038: Dietary Capsaicin May Decrease Blood Pressure Through Enhancing NO With eNOS Activation in 2-Kidney, 1-Clip Hypertensive Rats

Circulation ◽  
2015 ◽  
Vol 131 (suppl_1) ◽  
Author(s):  
Yukiko Segawa ◽  
Hiroko Hashimoto ◽  
Tomoko Osera ◽  
Nobutaka Kurihara
Keyword(s):  
Blood Pressure ◽  
Protein Expression ◽  
Mrna Expression ◽  
Dietary Therapy ◽  
Left Renal Artery ◽  
Sham Operation ◽  
Hypertensive Rats ◽  
Enos Mrna ◽  
Enos Protein Expression ◽  
Enos Protein

Objective: Capsaicin, a component of chili peppers, is reported to have beneficial effects on cardiovascular system through the vasodilative effects. We recently demonstrated the alleviation of blood pressure (BP) elevation by consuming a low concentration of capsaicin diet in 2-kidney, 1-clip (2K1C) hypertensive rats. Since the alleviation was diminished when 2K1C rats took NG-nitro-L-arginine methyl ester, a NO synthase (NOS) inhibitor, during the protocol, we hypothesized that NO has a key role in the effect of capsaicin in 2K1C rats. In this study, we observed eNOS mRNA expression and protein expressions of eNOS and phosphorylated eNOS in 2K1C rats fed a diet containing capsaicin. Methods: Six-week old male Sprague-Dawley rats were treated with sham operation (SHAM) or clipping the left renal artery (2K1C). One week after the surgery, each group of rats were further divided into 2 groups randomly, which received either a control diet (CTL) or a diet containing 0.006% capsaicin (CAP) for 6 weeks. The systolic BP was measured by a tail-cuff method once per week throughout the protocol. At the end of the protocol, rats were euthanized and the abdominal aortas were collected for extracting mRNA and protein. Then, the expression of eNOS mRNA and protein in aorta was evaluated in each group of rats by real time RT-PCR and Western blotting. Results: As shown in Table, capsaicin diet alleviated BP elevation in 2K1C rats. After the dietary protocol, eNOS mRNA expression in 2K1C-CAP was significantly higher than in 2K1C-CTL. Although there were no significant differences in eNOS protein expression among four groups, phosphorylated eNOS protein expression in 2K1C-CAP was marginally significantly higher than in 2K1C-CTL. The expression was also significantly higher in 2K1C rats than in SHAM. Discussion: The present data suggested that dietary capsaicin decreases BP through enhancing NO with activation of eNOS in 2K1C hypertensive rats. It may be a clue for developing a dietary therapy for prevention of hypertension.

scholarly journals Lobe-Specific Heterogeneity in Asymmetric Dimethylarginine and Matrix Metalloproteinase Levels in a Rat Model of Obstructive Cholestasis

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Andrea Ferrigno ◽  
Giuseppina Palladini ◽  
Alberto Bianchi ◽  
Vittoria Rizzo ◽  
Laura G. Di Pasqua ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Asymmetric Dimethylarginine ◽  
Left Lobe ◽  
Sham Operation ◽  
Hepatic Enzymes ◽  
Common Bile Duct Ligation ◽  
Nitric Oxide Synthase Inhibitor ◽  
Duct Ligation ◽  
Obstructive Cholestasis ◽  
Synthase Inhibitor

We investigated the effects of obstructive cholestasis in different hepatic lobes by evaluating asymmetric dimethylarginine (ADMA) (a nitric oxide synthase inhibitor), protein methyltransferase (PRMT) and dimethylarginine dimethylaminohydrolase (DDAH) (enzymes involved, resp., in its synthesis and degradation), the cationic transporter (CAT), and metalloproteinase (MMP) activity. Sixteen male Wistar rats underwent a 3-day cholestasis by common bile duct ligation (BDL) or sham operation. Blood samples and hepatic biopsies from left lobe (LL), median lobe (ML), and right lobe (RL) were collected. Serum hepatic enzymes, tissue ADMA, DDAH activity, CAT-2 protein, mRNA expression of DDAH and PRMT, and MMP-2 and MMP-9 activity were monitored. Cholestasis was confirmed by altered serum hepatic enzymes. Higher levels of tissue ADMA were detected in RL and ML as compared with LL. PRMT mRNA expression and DDAH activity did not differ among the lobes after BDL. CAT-2 levels are higher in the RL and ML in the sham-operated group. Higher activity in MMP-2 and MMP-9 was found in RL. In conclusion, after cholestasis an increase in hepatic ADMA in RL and ML was detected as well as tissue MMP-2 and MMP-9 activation in RL, supporting the evidence of functional heterogeneity among the liver lobes also occurring in an obstructive cholestasis model.

Role of heme oxygenase-carbon monoxide pathway in pathogenesis of cirrhotic cardiomyopathy in the rat

2001 ◽  
Vol 280 (1) ◽  
pp. G68-G74 ◽  
Author(s):  
Hongqun Liu ◽  
Daisheng Song ◽  
Samuel S. Lee
Keyword(s):  
Bile Duct ◽  
Protein Expression ◽  
Heme Oxygenase ◽  
Papillary Muscle ◽  
Cardiac Contraction ◽  
Zinc Protoporphyrin ◽  
Sham Operation ◽  
Muscle Contractility ◽  
Cirrhotic Cardiomyopathy ◽  
Duct Ligation

The enzyme heme oxygenase (HO), which exists in inducible (HO-1) and constitutive (HO-2) isoforms, degrades heme to biliverdin and CO. CO depresses cardiac contraction via cGMP. We aimed to clarify a possible role for the HO-CO pathway in the pathogenesis of cirrhotic cardiomyopathy in bile duct-ligated rats. Four weeks after bile duct ligation or sham operation, rat ventricles were examined for HO-1 and HO-2 mRNA by RT-PCR and for protein expression by Western blotting. Total HO enzyme activity and cGMP levels were also measured. The effects of a HO inhibitor, zinc protoporphyrin IX (ZnPP), on ventricular cGMP levels and isolated papillary muscle contractility were studied. We found that HO-1 mRNA transcription and protein expression were significantly augmented in cirrhotic hearts compared with sham-operated controls, whereas there was no difference in HO-2 mRNA or protein levels. Total HO activity and cGMP levels were significantly increased in cirrhotic ventricles vs. controls. In cirrhotic ventricles, treatment with ZnPP significantly decreased cGMP production and improved the blunted papillary muscle contractility, whereas it had no effect on control muscles. CO perfusion inhibited papillary muscle contractility, an effect completely blocked by methylene blue and partially blocked by ZnPP. These results indicate that activation of the HO-CO-cGMP pathway is involved in the pathogenesis of cirrhotic cardiomyopathy.

scholarly journals Conclusive evidence of endotoxaemia in biliary obstruction

Gut ◽  
1998 ◽  
Vol 42 (2) ◽  
pp. 293-299 ◽  
Author(s):  
W D B Clements ◽  
P Erwin ◽  
M D McCaigue ◽  
I Halliday ◽  
G R Barclay ◽  
...  
Keyword(s):  
Obstructive Jaundice ◽  
Biliary Obstruction ◽  
Inner Core ◽  
Humoral Response ◽  
Conclusive Evidence ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Method ◽  
Sham Operation ◽  
Strong Positive Correlation ◽  
Duct Ligation

Background—Endotoxaemia is implicated in the pathophysiology of obstructive jaundice. The EndoCab enzyme linked immunosorbent assay (ELISA) is a novel assay which measures endogenous antibody (IgG) to the inner core region of circulating endotoxins (ACGA).Aims—To investigate the significance of endotoxaemia in biliary obstruction using the EndoCab assay and assess the specificity of the humoral response to endotoxin compared with an exogenous antigenic challenge (tetanus toxoid, TT).Methods—Three groups of adult male Wistar rats were studied: no operation, sham operation, and bile duct ligation for 21 days (BDL). In the second study, rats rats received prior immunisation with TT.Results—In the preliminary experiment, plasma ACGA was significantly increased in the BDL group (306.6 (18.3)% versus 119.9 (6.7)% and 105.2 (4.6)% in the sham and no operation groups, respectively; p<0.001). Although the mean endotoxin concentration in the BDL group was greater than that in the control groups this was not significant. There was a strong positive correlation between ACGA and endotoxin concentrations (p=0.0021). In the second study mean ACGA after 21 days of BDL was significantly elevated (267.1 (31.2)% versus 101.6 (21.2)% at baseline, p<0.0001). ACGA was unaffected in the other two groups. TT antibody concentrations fell in all three groups; only in the BDL group was the fall significant (97.6 (5.3)% versus 78.8 (4.2)% at baseline, p<0.05).Conclusions—The specific rise in ACGA supports the hypothesis that endotoxin has an integral role in the pathophysiology of obstructive jaundice. The production of anticore glycolipid antibodies specifically reflects systemic endotoxaemia in this model. The EndoCab assay provides a novel, sensitive, and specific method for endotoxin detection.

scholarly journals Early type 1 diabetes aggravates renal ischemia/reperfusion-induced acute kidney injury

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mariana Charleaux de Ponte ◽  
Vanessa Gerolde Cardoso ◽  
Guilherme Lopes Gonçalves ◽  
Juliana Martins Costa-Pessoa ◽  
Maria Oliveira-Souza
Keyword(s):  
Renal Function ◽  
Protein Expression ◽  
Mrna Expression ◽  
Ischemia Reperfusion ◽  
Plasma Creatinine ◽  
Sham Operation ◽  
Tubular Injury ◽  
Early Diabetes ◽  
Gene And Protein Expression ◽  
Kidney Morphology

AbstractThe present study aimed to investigate the interaction between early diabetes and renal IR-induced AKI and to clarify the mechanisms involved. C57BL/6J mice were assigned to the following groups: (1) sham-operated; (2) renal IR; (3) streptozotocin (STZ—55 mg/kg/day) and sham operation; and (4) STZ and renal IR. On the 12th day after treatments, the animals were subjected to bilateral IR for 30 min followed by reperfusion for 48 h, at which time the animals were euthanized. Renal function was assessed by plasma creatinine and urea levels, as well urinary protein contents. Kidney morphology and gene and protein expression were also evaluated. Compared to the sham group, renal IR increased plasma creatinine, urea and albuminuria levels and decreased Nphs1 mRNA expression and nephrin and WT1 protein staining. Tubular injury was observed with increased Havcr1 and Mki67 mRNA expression accompanied by reduced megalin staining. Renal IR also resulted in increased SQSTM1 protein expression and increased proinflammatory and profibrotic factors mRNA expression. Although STZ treatment resulted in hyperglycemia, it did not induce significant changes in renal function. On the other hand, STZ treatment aggravated renal IR-induced AKI by exacerbating renal dysfunction, glomerular and tubular injury, inflammation, and profibrotic responses. Thus, early diabetes constitutes a relevant risk factor for renal IR-induced AKI.

Changes in G protein expression account for impaired modulation of hepatic cAMP formation after BDL

1998 ◽  
Vol 274 (6) ◽  
pp. G1151-G1159 ◽  
Author(s):  
Bernard Bouscarel ◽  
Yasushi Matsuzaki ◽  
Man Le ◽  
Thomas W. Gettys ◽  
Hans Fromm
Keyword(s):  
Bile Acid ◽  
G Protein ◽  
Receptor Expression ◽  
Sham Operation ◽  
Acid Uptake ◽  
Α Subunit ◽  
Camp Synthesis ◽  
Bile Acid Uptake ◽  
Duct Ligation ◽  
Camp Formation

The regulation of cAMP synthesis by hormones and bile acids is altered in isolated hamster hepatocytes 2 days after bile duct ligation (BDL) [Y. Matsuzaki, B. Bouscarel, M. Le, S. Ceryak, T. W. Gettys, J. Shoda, and H. Fromm. Am. J. Physiol. 273 ( Gastrointest. Liver Physiol. 36): G164–G174, 1997]. Therefore, studies were undertaken to elucidate the mechanism(s) responsible for this impaired modulation of cAMP formation. Hepatocytes were isolated 48 h after either a sham operation or BDL. Both preparations were equally devoid of cholangiocyte contamination. Although the basal cAMP level was not affected after BDL, the ability of glucagon to maximally stimulate cAMP synthesis was decreased by ∼40%. This decreased glucagon effect after BDL was not due to alteration of the total glucagon receptor expression. However, this effect was associated with a parallel 50% decreased expression of the small stimulatory G protein α-subunit (GsαS). The expression of either the large subunit (GsαL) or the common β-subunit remained unchanged. The expression of Giα2and Giα3was also decreased by 25 and 46%, respectively, and was associated with the failure of ANG II to inhibit stimulated cAMP formation. Therefore, alterations of the expression of GsαSand Giα are, at least in part, responsible for the attenuated hormonal regulation of cAMP synthesis. Because cAMP has been reported to stimulate both bile acid uptake and secretion, impairment of cAMP synthesis and bile acid uptake may represent an initial hepatocellular defense mechanism during cholestasis.

Induction of short heterodimer partner 1 precedes downregulation of Ntcp in bile duct-ligated mice

2002 ◽  
Vol 282 (1) ◽  
pp. G184-G191 ◽  
Author(s):  
Gernot Zollner ◽  
Peter Fickert ◽  
Dagmar Silbert ◽  
Andrea Fuchsbichler ◽  
Conny Stumptner ◽  
...  
Keyword(s):  
Steady State ◽  
Bile Acid ◽  
Bile Duct ◽  
Protein Expression ◽  
Mrna Expression ◽  
Bile Acids ◽  
Serum Bile Acid ◽  
Acid Uptake ◽  
Mrna Levels ◽  
Short Heterodimer Partner

Cholestasis is associated with retention of bile acids and reduced expression of the Na+/taurocholate cotransporter (Ntcp), the major hepatocellular bile acid uptake system. This study aimed to determine whether downregulation of Ntcp in obstructive cholestasis 1) is a consequence of bile acid retention and 2) is mediated by induction of the transcriptional repressor short heterodimer partner 1 (SHP-1). To study the time course for the changes in serum bile acid levels as well as SHP-1 and Ntcp steady-state mRNA levels, mice were subjected to common bile duct ligation (CBDL) for 3, 6, 12, 24, 72, and 168 h and compared with sham-operated controls. Serum bile acid levels were determined by radioimmunoassay. SHP-1 and Ntcp steady-state mRNA expression were assessed by Northern blotting. In addition, Ntcp protein expression was studied by Western blotting and immunofluorescence microscopy. Increased SHP-1 mRNA expression paralleled elevations of serum bile acid levels and was followed by downregulation of Ntcp mRNA and protein expression in CBDL mice. Maximal SHP-1 mRNA expression reached a plateau phase after 6-h CBDL (12-fold; P < 0.001) and preceded the nadir of Ntcp mRNA levels (12%, P < 0.001) by 6 h. In conclusion, bile acid-induced expression of SHP-1 may, at least in part, mediate downregulation of Ntcp in CBDL mice. These findings support the concept that downregulation of Ntcp in cholestasis limits intracytoplasmatic accumulation of potentially toxic bile acids.

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