Effect of Bile-Duct Ligation on Organelle Marker Enzymes in the Liver and Serum of Rats

1. Marker enzymes for the principal subcellular organelles of rat liver were asayed in the liver of rats 1 day and 8 days after bile-duct ligation or after laparotomy as a control procedure. 2. The microsomal enzymes in liver tissue showed complex changes. Benz[α]pyrene hydroxylase activity, predominantly found in the smooth endoplasmic reticulum, was decreased. Glucose 6-phosphatase activity and ribonucleic acid, which are localized predominantly in the rough endoplasmic reticulum, were increased. 3. The plasma membrane enzyme, alkaline phosphatase, increased in activity after bile-duct ligation. 4. No changes in mitochondrial enzyme activities were noted after 1 day but there was a 50% reduction 8 days after ligation. Lysosomal enzyme activities did not change in the liver tissue. 5. Liver catalase and d-amino acid oxidase activities showed a slight increase at 1 day post-ligation but a significant fall by 8 days. 6. Lactate dehydrogenase, a cytosol enzyme, showed a decrease in activity after 1 day but an increase in tissue activities 8 days after ligation. 7. Serum activities of mitochondrial, plasma membrane, microsomal, lysosomal and cytosol marker enzymes tended to increase post-ligation, particularly at 8 days. 8. Monoamine oxidase, a predominantly mitochondrial enzyme, was greatly elevated in the serum after 1 day but had returned to normal activities by 8 days.

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Plasma Membrane Form of Phosphatidate Phosphohydrolase: A Possible Role in Signal Transduction during Liver Fibrogenesis

Clinical Science ◽

10.1042/cs0850281 ◽

1993 ◽

Vol 85 (3) ◽

pp. 281-287 ◽

Cited By ~ 5

Author(s):

Christopher P. Day ◽

Alastair D. Burt ◽

Ashley Stj. M. Brown ◽

Mark K. Bennett ◽

Desmond J. Farrell ◽

...

Keyword(s):

Plasma Membrane ◽

Liver Disease ◽

Bile Duct ◽

Common Bile Duct ◽

Alcoholic Liver Disease ◽

Bile Duct Ligation ◽

Sham Operation ◽

Common Bile Duct Ligation ◽

Duct Ligation ◽

Phosphatidate Phosphohydrolase

1. Several growth factors important in liver regeneration and fibrosis stimulate phospholipase D in plasma membranes via a receptor/G-protein-coupled mechanism resulting in hydrolysis of phosphatidylcholine to phosphatidate. Phosphatidate can be further hydrolysed to diacylglycerol by phosphatidate phosphohydrolase. Phosphatidate and diacylglycerol can act as ‘second-messengers’ and regulation of phosphatidate phosphohydrolase activity could control the balance between them. 2. A form of phosphatidate phosphohydrolase, located in the plasma membrane and insensitive to inhibition by N-ethylmaleimide, has recently been identified that is distinct from the ‘metabolic’ form, which is present in the cytosol and microsomes and is sensitive to N-ethylmaleimide. 3. We have investigated the hypothesis that the balance between regeneration and fibrosis is, in part, determined by the activity of plasma membrane phosphatidate phosphohydrolase through its effect on the phosphatidate/diacylglycerol ratio. N-Ethylmaleimide-insensitive and -sensitive phosphatidate phosphohydrolase activities were measured in three hepatic conditions characterized by regeneration and/or fibrosis: alcoholic liver disease in humans (regeneration and fibrosis) and rat livers after either acute CCl-4-induced injury (regeneration) or common bile duct ligation (fibrosis). 4. In patients with alcoholic liver disease, N-ethylmaleimide-insensitive phosphatidate phosphohydrolase activity was higher in cirrhotic biopsies (5.82±0.3 nmol of Pi min−1 mg−1 of protein, n = 19) than in non-cirrhotic biopsies (2.17 ±0.2, n = 23) or in wedge biopsies from healthy subjects undergoing routine cholecystectomy (2.16 ±0.5, n = 6). N-Ethylmaleimide-insensitive phosphatidate phosphohydrolase activity was unchanged in the 10 days after CCl4 treatment but increased progressively in common bile duct-ligated rats (e.g. day 28: ‘sham’ operation, 1.97 ±0.3, chronic bile duct ligation, 6.91 ±1.24 nmol of Pi min−1 mg−1 of protein). N-Ethylmaleimide-insensitive phosphatidate phosphohydrolase activity correlated closely with the degree of fibrosis in humans and rats. N-Ethylmaleimide-sensitive phosphatidate phosphohydrolase activity was unchanged after CCI4 treatment or common bile duct ligation and was not increased in cirrhotic livers. 5. Plasma membrane N-ethylmaleimide-insensitive phosphatidate phosphohydrolase increases in liver fibrosis but not regeneration. Stimulation of phosphatidate phosphohydrolase activity with its effect on the diacylglycerol/phosphatidate ratio may play a role in transduction of the fibrosis signal.

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MicroRNA-29a mitigation of endoplasmic reticulum and autophagy aberrance counteracts in obstructive jaundice-induced fibrosis in mice

Experimental Biology and Medicine ◽

10.1177/1535370217741500 ◽

2017 ◽

Vol 243 (1) ◽

pp. 13-21 ◽

Cited By ~ 13

Author(s):

Ying-Hsien Huang ◽

Ya-Ling Yang ◽

Fu-Chen Huang ◽

Mao-Meng Tiao ◽

Yen-Cheng Lin ◽

...

Keyword(s):

Endoplasmic Reticulum ◽

Bile Duct ◽

Protein Expression ◽

Bile Duct Ligation ◽

Binding Protein ◽

Wild Type ◽

Innovative Strategy ◽

Over Expression ◽

Duct Ligation

Hepatic fibrosis was caused by a number of signaling pathways that damage liver integrity. We have previously shown that microRNA-29a (miR-29a) protects against liver fibrosis. Aberrant endoplasmic reticulum (ER) and autophagy function reportedly exaggerate hepatic disorders. The aim of this study was to characterize the biological influence of miR-29a on ER function in injured livers with bile duct ligation (BDL). We performed BDL on miR-29a transgenic mice (miR-29aTg) and wild-type mice to induce cholestatic liver injury. Rat T6 cells were transfected with miR-29a mimic and tunicamycin. Compared to the wild-type mice, the BDL deterioration of liver function in terms of total bilirubin, alanine transaminase, and aspartate transaminase activity in the miR-29aTg mice was significantly reduced. Affected livers in the miR-29aTg mice demonstrated a slight fibrotic matrix formation. miR-29a over-expression reduced the BDL disturbance of the expressions of inositol-requiring kinase 1alpha, double-stranded RNA-activated protein kinase-like endoplasmic reticulum kinase, spliced-X-box binding protein 1 (sXBP1), CCAAT/enhancer-binding protein homologous protein (CHOP), ULK, LC3BII, p62, and cleaved caspase-8, 9 and 3. In vitro, T6 cells exposed to tunicamycin by increasing abundances of CHOP, sXBP1, cleaved caspase-3, and LC3BII were diminished in the cell cultures transfected with the miR-29a mimic. On the other hand, we observed that miR-29a signaling protected liver tissues from BDL-mediated metabolic dysfunction and excessive fibrosis histopathology. This study provides new molecular insight into the miR-29a stabilization of ER integrity that slows the progression of cholestatic liver deterioration. Impact statement Long-term hepatic damage caused by hepatitis and cholestasis can accelerate fibrosis matrix over-production, which is a harmful process attributed to the dysregulation of a number of cellular and molecular events. The purpose of this study is to characterize the biological influence of miR-29a on endoplasmic reticulum (ER) function in bile duct ligation (BDL)-injured livers. To the best of our knowledge, this report is the first demonstration that miR-29a over-expression diminishes BDL provocation of ER stress (unfolded protein response, UPR) effector protein expression. This work also demonstrates that miR-29a decreased caspases protein expression in cholestatic livers, while an increase in miR-29a function reduced sXBP1 and CHOP expressions in T6 cells in mice. Analyses of this study highlight that controlling miR-29a signaling can serve as an innovative strategy in the future for microRNA regulation of ER homeostasis to combat cholestasis induction hepatic disorders.

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Alterations in key carbohydrate-metabolizing enzyme activities in rat livers following bile duct ligation and its release

The Japanese Journal of Surgery ◽

10.1007/bf02470849 ◽

1988 ◽

Vol 18 (1) ◽

pp. 68-76 ◽

Cited By ~ 1

Author(s):

Kenichi Hirata ◽

Hisashi Mimura ◽

Kunzo Orita

Keyword(s):

Bile Duct ◽

Enzyme Activities ◽

Bile Duct Ligation ◽

Duct Ligation ◽

Metabolizing Enzyme ◽

Rat Livers

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Identification of B10, an alkaline phosphodiesterase of the apical plasma membrane of hepatocytes and biliary cells, in rat serum: Increased levels following bile duct ligation and during the development of cholangiocarcinoma

Hepatology ◽

10.1002/hep.510270234 ◽

1998 ◽

Vol 27 (2) ◽

pp. 563-568 ◽

Cited By ~ 19

Author(s):

Nirina Rajho Meerson ◽

Dani�le Delautier ◽

Anne-Marie Durand-Schneider ◽

Alain Moreau ◽

Michael L. Schilsky ◽

...

Keyword(s):

Plasma Membrane ◽

Bile Duct ◽

Bile Duct Ligation ◽

Apical Plasma Membrane ◽

Rat Serum ◽

Duct Ligation

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The Protective Effects of Magnesium on Liver Tissue Alteration Induced by Bile Duct Ligation in Rat

International Journal of Advances in Chemical Engineering and Biological Sciences ◽

10.15242/ijacebs.a0915005 ◽

2015 ◽

Vol 2 (1) ◽

Cited By ~ 1

Keyword(s):

Bile Duct ◽

Liver Tissue ◽

Bile Duct Ligation ◽

Protective Effects ◽

Duct Ligation ◽

Tissue Alteration

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CYTOCHEMISTRY OF GOLGI FRACTIONS PREPARED FROM RAT LIVER

The Journal of Cell Biology ◽

10.1083/jcb.60.1.8 ◽

1974 ◽

Vol 60 (1) ◽

pp. 8-25 ◽

Cited By ~ 164

Author(s):

Marilyn G. Farquhar ◽

J. J. M. Bergeron ◽

George E. Palade

Keyword(s):

Endoplasmic Reticulum ◽

Plasma Membrane ◽

Acid Phosphatase ◽

Liver Tissue ◽

Marker Enzymes ◽

Reliable Identification ◽

Fractionation Procedure ◽

Thiamine Pyrophosphatase ◽

Opposite Face ◽

Liver Homogenates

Cytochemical tests for several marker enzymes were applied to liver tissue and to the three Golgi fractions (GF1, GF2, GF3) separated by the procedure of Ehrenreich et al. from liver homogenates of alcohol-treated rats. 5'-Nucleotidase (AMPase) reaction product was found in all three fractions but in different locations: It occurred along the inside of the membrane of VLDL-filled vacuoles in GF1 and GF2, and along the outside of the cisternal membranes in GF3. In the latter it was restricted to the dilated cisternal rims and was absent from the cisternal centers. The AMPase activity found in the fractions by biochemical assay is therefore indigenous to Golgi components and is not due to contamination by plasma membrane. Acid phosphatase (AcPase) reaction product was detected within lysosomal contaminants in GF1 and within many VLDL-filled vacuoles in GF1 and GF2, indicating that AcPase activity is due not only to contaminating lysosomes, but also to enzyme indigenous to Golgi secretory vacuoles. G-6-Pase reaction product was present in GF3 and within contaminating endoplasmic reticulum fragments, but not in other fractions. Thiamine pyrophosphatase (TPPase) was localized to some of the VLDL-filled vacuoles and cisternae in GF1 and GF2, and was not found in the cisternae in GF3. The results demonstrate the usefulness of cytochemical methods in monitoring the fractionation procedure: They have (a) allowed a reliable identification of contaminants, (b) made possible a distinction between indigenous and contaminating activities, and (c) shown, primarily by the results of the TPPase test, that the procedure achieves a meaningful subfractionation of Golgi elements, with GF1 and GF3, representing primarily trans-Golgi elements from the secretory Golgi face, and GF3 consisting largely of cis-Golgi components from the opposite face.

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