Multiorgan chronic inflammatory hepatobiliary pancreatic murine model deficient in tumor necrosis factor receptors 1 and 2.
OBJECTIVE
To provoke persistent/chronic multiorgan inflammatory response and to contribute to stones formation followed by fibrosis in hepatobiliary and pancreatic tissues.


METHODS
Tumor necrosis factor receptors 1 and 2 (TNFR1/R2) deficient mice reared in-house were given dibutyltin dichloride (DBTC) twice within 10 d by oral gavage delivery. Sham control animals received vehicle treatment and naïve animals remained untreated throughout the study. Animals were monitored daily for symptoms of pain and discomfort. The abdominal and hindpaw hypersensitivity were assessed with von Frey microfilaments. Exploratory behaviors were recorded at the baseline, after initiation of treatment, and before study termination. Histopathological changes were examined postmortem in tissues. Collagen accumulation and fibrosis were confirmed with Sirius Red staining.


RESULTS
Animals lost weight after oral administration of DBTC and developed persistent inflammatory abdominal and hindpaw hypersensitivity compared to sham-treated controls (P < 0.0001). These pain related secondary mechanical hypersensitivity responses increased more than 2-fold in DBTC-treated animals. The drastically diminished rearing and grooming rates persisted after DBTC administration throughout the study. Gross as well as micropathology at one month confirmed that animals treated with DBTC developed chronic hepatobiliary injuries evidenced with activation of stellate cells, multifocal necrosis, fatty degeneration of hepatocytes, periportal infiltration of inflammatory cells, and prominent biliary ductal dilation. The severity of hepatitis was scored 3.7 ± 0.2 (severe) in DBTC-treated animals vs score 0 (normal) in sham-treated animals. Fibrotic thickening was extensive around portal ducts, in hepatic parenchyma as well as in lobular pancreatic structures and confirmed with Sirius Red histopathology. In addition, pancreatic microarchitecture was presented with distortion of islets, and parenchyma, infiltration of inflammatory cells, degeneration, vacuolization, and necrosis of acinar cells and distention of pancreatic ducts. Extent of pancreatic damage and pancreatitis were scored 3.6 ± 0.4 (severe) for DBTC-treated in contrast to score 0 (normal) in sham-treated animals. The gall bladder became expanded with ductal distention, and occasional bile stones were detected along with microscopic hepatic lesions. DBTC-treated animals developed splenic hypertrophy with increased weight and length (P < 0.01) along with thymic atrophy (P < 0.001). Finally, colitic lesions and colitis were prominent in DBTC-treated animals and scored 3.4 ± 0.3 (moderately severe) vs 0 (normal) for the sham-treated animals.


CONCLUSIONS
This is the first report of chronic inflammatory multiorgan hepatobiliary pancreatitis, along with fibrosis and calculi formation induced reliably utilizing oral DBTC administration in TNFR1/R2 deficient mice.
Helieh S Oz, Department of Physiology and Internal Medicine, College of Medicine, University of Kentucky Medical Center, Lexington, KY 40536, United States.
Helieh S
HS
Oz
D015746:Abdominal Pain / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001522:Behavior, Animal; D001652:Bile Ducts / Q000378:metabolism* / Q000473:pathology; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology / Q000235:genetics / Q000378:metabolism* / Q000523:psychology; D002761:Cholangitis / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism* / Q000523:psychology; D003092:Colitis / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism; D005106:Exploratory Behavior; D020022:Genetic Predisposition to Disease; D006120:Grooming; D055166:Hepatic Stellate Cells / Q000378:metabolism / Q000473:pathology; D006930:Hyperalgesia / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism; D020347:Lithiasis / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism* / Q000523:psychology; D008099:Liver / Q000378:metabolism* / Q000473:pathology; D008106:Liver Cirrhosis, Experimental / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism* / Q000523:psychology; D018345:Mice, Knockout; D009947:Organotin Compounds; D058748:Pain Perception; D010179:Pancreas / Q000378:metabolism* / Q000473:pathology; D058954:Pancreatic Stellate Cells / Q000378:metabolism / Q000473:pathology; D010195:Pancreatitis / Q000235:genetics / Q000378:metabolism* / Q000523:psychology; D010641:Phenotype; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000172:deficiency* / Q000235:genetics; D047889:Receptors, Tumor Necrosis Factor, Type II / Q000172:deficiency* / Q000235:genetics; D013154:Spleen / Q000378:metabolism / Q000473:pathology; D015431:Weight Loss
D009947:Organotin Compounds; D047888:Receptors, Tumor Necrosis Factor, Type I; D047889:Receptors, Tumor Necrosis Factor, Type II; C506236:Tnfrsf1a protein, mouse; C056189:di-n-butyltin
Calculi formation; Gall bladder; Hepatitis; Hepatobiliary inflammation; Inflammatory pain; Multiorgan; Pancreatitis