Laser enhancement in hepatic regeneration for partially hepatectomized rats.

BACKGROUND AND OBJECTIVE: The bio-stimulation effect of laser has been observed in many areas of Medicine. However, there are a few works which investigate its use for liver regeneration. Most of their results were inconclusive due to the use of high power lasers. This work was carried out to investigate the bio-stimulation effect of laser in liver regeneration using low power lasers. STUDY DESIGN/MATERIALS AND METHODS: We used Wistar male rats, which were irradiated with laser light (wavelength 590 nm and intensity of 50 mW/cm(2)) for 5 minutes after 70% hepatectomy. The respiratory mitochondrial activity, the serum level of aminotransferase and the PCNA were measured. RESULTS: Our results show a dramatic increase in the mitochondrial activity for the laser treated group at 24 hours after the hepatectomy. CONCLUSIONS: We conclude that the laser promotes a bio-stimulation effect on the early stages of liver regeneration without any detectable damage of the cells.

Angiotensin-converting enzyme inhibition by lisinopril enhances liver regeneration in rats.

Bradykinin has been reported to act as a growth factor for fibroblasts, mesangial cells and keratinocytes. Recently, we reported that bradykinin augments liver regeneration after partial hepatectomy in rats. Angiotensin-converting enzyme (ACE) is also a powerful bradykinin-degrading enzyme. We have investigated the effect of ACE inhibition by lisinopril on liver regeneration after partial hepatectomy. Adult male Wistar rats underwent 70% partial hepatectomy (PH). The animals received lisinopril at a dose of 1 mg kg body weight(-1) day(-1), or saline solution, intraperitoneally, for 5 days before hepatectomy, and daily after surgery. Four to six animals from the lisinopril and saline groups were sacrificed at 12, 24, 36, 48, 72, and 120 h after PH. Liver regeneration was evaluated by immunohistochemical staining for proliferating cell nuclear antigen using the PC-10 monoclonal antibody. The value for the lisinopril-treated group was three-fold above the corresponding control at 12 h after PH (P<0.001), remaining elevated at approximately two-fold above control values at 24, 36, 48 (P<0.001), and at 72 h (P<0.01) after PH, but values did not reach statistical difference at 120 h after PH. Plasma ACE activity measured by radioenzymatic assay was significantly higher in the saline group than in the lisinopril-treated group (P<0.001), with 81% ACE inhibition. The present study shows that plasma ACE inhibition enhances liver regeneration after PH in rats. Since it was reported that bradykinin also augments liver regeneration after PH, this may explain the liver growth stimulating effect of ACE inhibitors.

Angiotensin-converting enzyme inhibition by lisinopril enhances liver regeneration in rats

Bradykinin has been reported to act as a growth factor for fibroblasts, mesangial cells and keratinocytes. Recently, we reported that bradykinin augments liver regeneration after partial hepatectomy in rats. Angiotensin-converting enzyme (ACE) is also a powerful bradykinin-degrading enzyme. We have investigated the effect of ACE inhibition by lisinopril on liver regeneration after partial hepatectomy. Adult male Wistar rats underwent 70 percent partial hepatectomy (PH). The animals received lisinopril at a dose of 1 mg kg body weight-1 day-1, or saline solution, intraperitoneally, for 5 days before hepatectomy, and daily after surgery. Four to six animals from the lisinopril and saline groups were sacrificed at 12, 24, 36, 48, 72, and 120 h after PH. Liver regeneration was evaluated by immunohistochemical staining for proliferating cell nuclear antigen using the PC-10 monoclonal antibody. The value for the lisinopril-treated group was three-fold above the corresponding control at 12 h after PH (P<0.001), remaining elevated at approximately two-fold above control values at 24, 36, 48 (P<0.001), and at 72 h (P<0.01) after PH, but values did not reach statistical difference at 120 h after PH. Plasma ACE activity measured by radioenzymatic assay was significantly higher in the saline group than in the lisinopril-treated group (P<0.001), with 81 percent ACE inhibition. The present study shows that plasma ACE inhibition enhances liver regeneration after PH in rats. Since it was reported that bradykinin also augments liver regeneration after PH, this may explain the liver growth stimulating effect of ACE inhibitors

Efeitos do pré-condicionamento hepático em ratos cirróticos, submetidos à isquemia e reperfusäo hepática: resultados preliminares / Effects of the hepatic preconditioning in rats with liver cirrhosis, submitted to ischemia of the hepatic reperfusion: preliminary result

Baseando-se nos efeitos estimuladores do metabolismo energético pelo pré-condicionamento isquêmico (PCI) no tecido hepático, estudou-se dois grupos de ratos cirróticos submetidos a isquemia de 20 min e reperfusäo de 120 min, após o PCI ou näo respectivamente, determinando assim o valor do seu uso no prolongamento da manobra de Pringle e na regeneraçäo hepática na hepatectomia. Descritores: Cirrose hepática; isquemia hepática; razäo de controle respiratório(RCR); pré-condicionamento isquêmico hepático.

Evoluçäo da fibrose biliar secundária em ratos tratados mediante derivaçäo bílio duodenal ou bíliojejunal com alça de roux medindo 5, 10 e 15 cm / Evolution of secondary biliary fibrosis in rats treated with bile-duodenal shunt or roux-en-y bile-jejunal shunt measuring 5, 10 and 15 centimeters

A modalidade de derivaçäo bílio-digestiva empregada no tratamento da colestase extra-hepática crônica pode influenciar na reparaçäo das lesöes hepáticas. Avaliou-se o desempenho das derivaçöes bílio-duodenal e bílio-jejunal em Y de Roux com alça exclusa de diferentes comprimentos na reparaçäo das lesöes morfológicas e funcionais do fígado de ratos com fibrose biliar secundária. Foram utilizados ratos Wistar, com 15 dias de obstruçäo biliar, alocados em 5 grupos de 6 animais. O grupo OB caracterizou as alteraçöes da fibrose biliar. Os animais remanescentes foram tratados mediante derivaçäo com o duodeno (grupo DBD), e com o jejuno, em alça exclusa de 5cm (grupo DBJ5), 10cm (grupo DBJ10) e 15cm (grupo DBJ15), sendo reavaliados 3 meses depois. Outros 6 animais foram submetidos à inteTodos animais foram submetidos à avaliaçäo morfométrica do fígado, análise bioquímica do sangue e microbiológica da bile, estudo da funçäo mitocondrial hepática e verificaçäo do peso úmido do fígado e do baço. Na análise estatística adotou-se o nível de significância de 5 por cento. Houve aumento significativo do peso estimado, em g/Kg de peso corporal, dos ductos biliares, da fibrose e dos hepatócitos nos animais do grupo OB (medianas de 1,30; 10,03 e 37,0) em relaçäo aos animais controles (IS) (medianas de 0,03; zero e 29,37). Após tratamento, ocorreu regressäo significativa do peso estimado dos ductos biliares e da fibrose, com valores medianos de 0,22 e 0,22 para o grupo DBD, 0,45 e 3,31 para o grupo DBJ5 e 0,22 e 5,0 para o grupo DBJ15. Houve regressäo significativa do peso estimado dos hepatócitos apenas nos grupos derivados com o jejuno, com valores medianos de 31,93; 24,46 e 28,52. Ocorreu aumento significativo do peso úmido do fígado e do baço no grupo OB (medianas em g/Kg de peso corporal de 49,85 e 5,71) em relaçäo ao grupo IS (30,0 e 3,04). Houve regressäo significativa do peso do fígado em todos os tratamentos e do peso do baço nos animais tratados com derivaçäo bílio-jejunal, (valores medianos de 35,59 e 2,53 para DBJ5, 37,54 e 2,82 para DBJ10 e 32,73 e 2,93 para DBJ15). Após o tratamento, surgiram infiltrado inflamatório misto, nos espaços portais, refluxo enterobiliar e contaminaçäo bacteriana da bile. Houve aumento significativo no consumo de oxigênio pela mitocôndrias hepáticas nos estados 3 e 4 no grupo OB (medianas de 101,55 e 31,05 nanoátomos de O2/mgprot./min), em relaçäo ao grupo IS (medianas de 57,22 e 15,51)...

Biochemical and morphological changes in the liver after hepatic artery ligation in the presence or absence of extrahepatic cholestasis.

The present study was carried out to investigate the biochemical and morphological changes in the liver after ligation of the hepatic artery (HA) in the presence and in the absence of extrahepatic cholestasis (EHC). The study was conducted on 100 rats divided into four groups of 25 animals each: group 1, sham operation; group 2, hepatic artery ligation (HAL); group 3, bile duct ligation (BDL); and group 4, HAL plus BDL. All animals were sacrificed 7 days after surgery when total bilirubin and fractions, alkaline phosphatase (AP), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured in serum and on the inner hepatocyte mitochondrial membrane (IHMM); the incidence of necrosis and the volume fractions of vessels, bile ducts and hepatocytes in the liver were also determined. HAL reduces the relative volumes of bile ducts, with no changes in levels of bilirubin and fractions, AP, ALT, AST and IHMM, but HAL associated with EHC reduces duct proliferation and the liver becomes more vulnerable to necrosis. In conclusion, the normal liver depends on HA flow and this dependence is more evident in the presence of EHC.

Lack of control of liver gluconeogenesis in cholestatic rats with reduced portal blood flow.

Previous studies indicated a role for ischemia in the metabolic changes induced by cholestasis. Liver pyruvate kinase is a key enzyme for the concurrent control of glycolysis and gluconeogenesis. In this experiment the control of pyruvate kinase activity was investigated in cholestatic rats. Pyruvate kinase kinetics changed from a sigmoidal type in sham-operated rats to a hyperbolic type in obstructed rats. The change in the enzymatic kinetics paralleled the reduction in the portal blood flow, which reached 50% of the control value 7 days after obstruction. Dibutyryl cyclic AMP (5 mumol/kg body wt) plus theophylline 0.1 mmol/L failed to inactivate the enzyme when injected into the portal veins of rats whose livers were obstructed 7 days before. Both the kinetics changes and the lack of phosphorylation control are compatible with ischemia.

Hepatic morphology changes after hepatic artery ligation in extrahepatic cholestasis.

The present study was performed to correlate changes in hepatic morphology with hepatic artery ligation (HAL) in the presence of extrahepatic cholestasis (EHC). The study was conducted on 36 male Wistar rats weighing 350 to 400 g, divided at random into four groups of 9 animals each: group I, sham operation (control); group II, HAL; group III, bile duct ligation (BDL); group IV, HAL plus BDL. After seven days, liver fragments were obtained for morphological study. The relative volume of the bile ducts was I > II < III and III > IV (P < 0.05). These data indicate that arterial irrigation is important for the nutrition of the biliary tree. Seventy-six percent of the animals submitted to HAL plus BDL showed hepatic necrosis. In general, the liver probably becomes more dependent on HA flow in the presence of EHC.

Hepatic morphology changes after hepatic artery ligation in extrahepatic cholestasis

The present study was performed to correlate changes in hepatic morphology with hepatic artery ligation (HAL) in the presence of extrahepatic cholestasis (EHC). The study was conducted on 36 male Wistar rats weighing 350 to 400 g, divided at random into four groups of 9 animals each: group I, sham operation (control); group II, HAL; group III, bile duct ligation (BDL); group IV, HAL plus BDL. After seven days, liver fragments were obtained for morphiological study. The relative volume of the bile ducts was I>IIIV (P<0.05). These data indicate that arterial irrigation is important for the nutrition of the biliary tree. Seventy-six percent of the animals sumitted to HAL plus BDL showed hepatic necrosis. In general, the liver propably becomes more dependent on HA flow in the presence of EHC

Integrity of liver inner mitochondrial membrane in rats with extrahepatic cholestasis.

To investigate the effect of extrahepatic cholestasis on integrity of the inner mitochondrial membrane, a study was conducted on two groups of rats: sham-operated control animals (N = 10) and rats subjected to extrahepatic cholestasis (EHC, N = 10) by double ligation of the hepatic duct. The animals were observed for 7 days and then sacrificed. The EHC group presented significantly higher serum levels of alanine aminotransferase, total bilirubins and alkaline phosphatase than the controls (P less than 0.01). Basal mitochondrial respiration (state IV), analyzed separately using either alpha-ketoglutarate or alpha-ketoglutarate + pyruvate as substrates, was similar in the two groups (P greater than 0.01). ADP-activated respiration, state III, diminished significantly in the EHC group. The results show that the decrease in mitochondrial function that has been reported by several investigators to occur in EHC is due to mitochondrial alterations not related to the ability of these organelles to maintain the proton gradient, since the inner mitochondrial membrane continued to be energized throughout the observation period.

Integrity of liver inner mitochondrial membrane in rats with extrahepatic cholestasis

To investigate the effect of extrahepatic cholestasis on integrity of the inner mitochondrial membrane, a study was conduced on two groups of rats: sham-operated control animals (N=10) and rats subjected to extrahepatic cholestasis (EHC, N+10) by double ligation of the hepatic duct. The animals were observed for 7 days and then sacrificed. The EHC group presented significantly hugher serum levels of alanine aminotransferase, total bilirubins and alkakine phosphatase than the controls (P<0.01). Basal mitochondrial respiration (state IV), analyzed separately using either *-ketoglutarate or *-ketoglutarate+ pyruvate as substrates, was similar in the two groups (P>0.01). ADP-activated respiration, state III, dimished significantly in the EHC group. The results show that the decrease in mitochondrial function that has been reported by several investigators to occur in EHC is due to mitochondrial a alterations not related to the ability of these organelles to maintain the proton gradient, since the inner mitochondrial membrane continued to be energized throughtout the observation period

Hepatic regeneration after hepatectomy in schistosomatic fibrosis.

After partial hepatectomy (approximately 30%), the labelled hepatocyte index (LHI) of the normal mouse is increased and reaches its highest value 48 h after surgery. In mice with schistosomatic fibrosis (SF), the LHI was lower than in the controls at 24, 48 and 72 h after hepatectomy.

Serum gamma-glutamyl transpeptidase in chronic extrahepatic cholestasis.

Studies in the literature have shown the inhibitory action of bilirubin on serum gamma-glutamyl transpeptidase (GGT) activity. In spite of this, GGT is a sensitive diagnostic indicator, comparable to alkaline phosphatase for the diagnosis of chronic extrahepatic cholestasis (CEHC). To evaluate the relationship of bilirubin, alkaline phosphatase (AP) activity and GGT activity in CEHC we studied 30 patients with CEHC divided into two groups: A, 14 male and female subjects with serum bilirubin levels of 2 to 10 mg/dl, and B, 16 male and female subjects with serum bilirubin levels of 10.1 to 44 mg/dl. Serum GGT activity was measured by diluting serum samples 1:5 and 1:10. GGT values were equivalent in the two groups, there was no correlation between bilirubin and GGT (r = +0.179, P less than 0.01), and there was a significant correlation between GGT and AP (r = +0.627, P less than 0.01). The present results show that, if GGT inhibition by bilirubin did occur, this inhibition was not sufficient to prevent the increase in serum GGT levels and that the sensitivity of GGT as a marker for CEHC is similar to that of alkaline phosphatase.

Inhibitory effect of different anticoagulants on gamma-glutamyl transpeptidase activity.

Increased gamma-glutamyl transpeptidase (GGT) levels have been observed in patients with several types of liver diseases. However, since GGT activity can be inhibited or increased by several substances, the interpretation of these increased levels should be made with caution. The present study was designed to determine plasma GGT activity in blood samples obtained from 24 adult volunteers (13 males and 11 females aged 20 to 40 years) with the use of three different anticoagulants, i.e., ethylenediaminetetraacetic acid (EDTA), calcium fluoride and calcium citrate, and to compare it with GGT activity in serum obtained from the same blood samples. Serum GGT activity was significantly higher than plasma GGT activity (P less than 0.001) and significantly higher in men than in women (P less than 0.05), though plasma GGT activity was similar for both sexes.

Serum transaminase levels in the acute phase of chronic extrahepatic cholestasis.

Serum alanine aminotransferase (ALT) and serum aspartate aminotransferase (AST) levels are normal or discretely increased in rats with chronic extrahepatic cholestasis (CEHC). During the acute phase (first 72 h after biliary obstruction), however, serum transaminase values are quite elevated due to a mechanism not yet fully elucidated. Thus, this is a good experimental model, not involving hepatocellular necrosis, for the study of serum ALT and AST levels during the acute phase of CEHC. Male Wistar rats (250-350 g) were divided into two groups: group A (N = 60) was submitted to sham operation for bile duct ligation (BDL), and group B (N = 60) was submitted to BDL. Thirty and 120 min after BDL there was a 1.5-fold increase in both serum ALT and AST levels compared to sham-operated rats (P less than 0.05). Serum ALT levels were higher than AST levels as early as 30 min after BDL and the highest serum values for both transaminases were observed at 360 min which was also the last value measured. Serum AST levels increased 120 min after BDL, with no further significant increases thereafter.

Effect of hepatic artery ligation in rats with chronic extrahepatic cholestasis.

Because of the liver's dependence on arterial blood to exert its metabolic functions in cirrhosis of the liver, with or without thrombosis of the portal vein, the interruption of hepatic arterial flow for the palliative treatment of malignant tumors of the liver is counterindicated. However, the effects of arterial devascularization on the cholestatic liver are not fully understood. The objective of the present study was to investigate hepatic alterations due to hepatic artery ligation in rats with chronic extrahepatic cholestasis. Serum alkaline phosphatase, bilirubin and alanine aminotransferase were measured in rats 3 h after sham operation (group A, N = 29) or ligation of the hepatic artery (group B, N = 29). Alanine aminotransferase activity was significantly higher (P less than 0.05) in group B, demonstrating acute hepatocellular damage in animals with chronic extrahepatic cholestasis.

Heparin-induced increase in plasma and serum gamma-glutamyl transpeptidase activity.

gamma-Glutamyl transpeptidase (GGT) activity may be a sensitive index of hepatobiliary dysfunction. GGT activity, however, has been shown to be affected by several factors in different clinical situations. The present study was designed to determine the effect of heparin on serum and plasma GGT activity. Blood samples were collected from 15 normal male Wistar rats weighing 200-250 g for the determination of GGT activity in plasma prepared with heparin (group P(H), N = 10), in serum (group S, N = 15) and in serum with heparin added (group S + H, N = 10). GGT activity was 45.1 +/- 9.5 U/l (mean +/- SD) for serum compared with 161.2 +/- 46.1 U/l for serum plus heparin and 93.3 +/- 30.9 U/l for plasma prepared with heparin. The mean for each group was significantly different from the means for the other groups. These data demonstrate that GGT activity measurements should be made on serum in the absence of heparin, which produces elevated results.

Lack of correlation between the sulfobromophthalein test and fatty liver in diabetic dogs.

The present study was performed to evaluate the correlation between fatty liver and liver function in diabetic dogs. Experimental diabetes was induced in 14 dogs by total pancreatectomy or by alloxan administration, and partial (30%) hepatectomy was performed. Diabetic dogs were compared with control dogs (N = 7) subjected only to partial hepatectomy (30%). Control and diabetic dogs were submitted to the sulfobromophthalein (BSP) liver function test. Serum BSP retention was increased in diabetic dogs but there was no correlation with the extent of fatty infiltration of the liver.

Effect of chlorpromazine and biliary drainage on portal blood flow and mitochondrial function during extrahepatic cholestasis.

The present study examines the effect of chlorpromazine and biliary drainage in cholestatic rats. The time course of portal blood flow was studied 24, 48, and 72 h and seven days after bile duct ligation. Portal blood flow decreased after 72 h. Chlorpromazine reduced biliary hydrostatic pressure in sham-operated control rats, but 24-h obstruction was sufficient to prevent this effect in cholestatic rats. The drug ameliorated the mitochondrial and cell membrane function of cholestatic rats before and after drainage. The data present further support for the role of ischemia in cholestasis.

Effect of hepatic artery ligation on liver metabolism in rats.

The rat is an experimental model of orthotopic liver transplant that does not need arterial revascularization of the transplanted liver. The objective of the present study was to investigate the effect of hepatic artery ligation on hepatic metabolism in rats. After 2, 3 or 24 h of hepatic artery ligation, no significant differences in serum levels of alanine aminotransferase were observed. Bile flow decreased significantly 3 h after arterial ligation (P less than 0.05) and returned to normal after 24 h (P less than 0.05).