Heterogeneity of rabbit hepatocytes for bile secretion after acinar zone 3 damage induced by bromobenzene. Effect of bilirubin and bile salt infusions

Anaesthetized rabbits were used to study the effect of bromobenzene-induced hepatic damage to the acinar zone 3 on bile flow, bile salt, sodium secretion as well as bilirubin transport in basal conditions or with infusion of sodium glycodeoxycholate. The bromobenzene-pretreated animals exhibited in basal conditions a lower bile flow (44%) than that of the controls, with a smaller decrease in bile salt output (27%) and sodium output (29%), whereas no modification in endogenous bilirubin excretion was observed. The bile salt independent fraction of secretion (BSIF) was reduced significantly after the toxic lesion both in terms of absolute and relative values. The hepatocytes of the periportal zone were capable of excreting the totality of bilirubin presented to the liver, regardless of the extent of bile flow or the input of bile salts. The infusion of bilirubin at 1.0 mumole/kg/min led to a fall in bile flow which was attributed to the interference of the pigment with the BSIF. The maximal bilirubin excretion was significantly smaller in bromobenzene-pretreated animals than in the controls, which could be due to the incapacity of the intoxicated rabbits to recruit quiescent hepatocytes. When glycodeoxycholate was administered under conditions of maximal bilirubin transport, bile flow increased as did bile salt secretion in both controls and animals with damaged livers. However, clear differences persisted between the two, which could be attributed not only to the volume fraction of necrosis but also to an interference by bilirubin with the hepatic handling of bile salts. Maximal bilirubin excretion increased in a similar way in both groups after glycodeoxycholate administration. It is proposed that glycodeoxycholate infusion facilitates the hepatic depletion of bilirubin, probably by stimulating transport processes.     

Effectiveness of sodium selenite in experimental liver dystrophy]

The effect of sodium selenite on the cholepoietic function of the liver in rats with acute dystrophy induced by carbon tetrachloride was studied. When used in doses of 1 and 10 gamma/100 g the drug was found to normalize the intensity of bile secretion, synthesis and secretion of bile acids and that of bilirubin, as well as excretion of cholesterol. This was attended by a rise of the cholate-cholesterol ratio.     

健脾消胀片对大鼠胆汁分泌的影响

目的：观察健脾消胀片对大鼠胆汁分泌的影响。方法：大鼠60只,分为6组,分别灌服健脾消胀片（大、中、小剂量）混悬液;舒胆通混悬液;清肝利胆胶囊混悬液和同体积的0.5%羧甲基纤维素;每天给药1次,连续给药5 d。收集各组大鼠胆汁量,检测胆汁中总胆汁酸、总胆固醇、总胆红素等含量。结果：健脾消胀片可使大鼠胆汁中胆汁酸、胆红素含量显著升高（P〈0.01）,胆固醇含量显著降低（P〈0.01）。结论：健脾消胀片有促进胆汁分泌的作用。     

Effects of ursodeoxycholate on maximal biliary secretion of bilirubin in the rat

The effect of sodium ursodeoxycholate (0.5 and 1.0 mumol/min/100 g) on the maximal biliary secretion (Tm) of bilirubin and on the concentration of bilirubin in liver and plasma at the end of a bilirubin load was studied in Wistar rats. Administration of ursodeoxycholate at 0.5 mumol/min/100 g caused a 0.8-fold increase in bile flow and a significant increase in the bilirubin Tm (+24%). This was associated with a significant reduction of liver and plasma bilirubin concentrations (-16% and -17%, respectively). Bilirubin UDP-glucuronosyltransferase activity was not significantly enhanced. There was a significant increase in the biliary excretion of bilirubin conjugates (+30%) and in the diconjugates/monoconjugates ratio in bile (+31%). When ursodeoxycholate was given at 1.0 mumol/min/100 g, it produced a 1.7-fold increase in bile flow, but the bilirubin Tm was significantly reduced (-21%). Liver bilirubin concentrations were decreased (-20%) and there was a significant enhancement in total pigment concentration in plasma (+19%). Both the excretion of unconjugated bilirubin and that of bilirubin conjugates were significantly reduced (-60% and -18%, respectively). There was a significant decrease in the bilirubin-UDP glucuronosyltransferase activity and the diconjugates/monoconjugates ratio in bile (-27% and -27%, respectively). These results indicate that ursodeoxycholate is able to increase maximal bilirubin secretion only when administered at low doses and that infusion at higher rates can significantly interfere with different steps in the hepatobiliary transport of the pigment.     

The effect of oral contraceptive steroids on bile secretion and bilirubin Tm in rats

1. The effect of oestrogens and progestogens and their 17alpha-ethinyl derivatives on bile flow, maximum rate of bilirubin secretion, serum and liver bilirubin has been studied.2. Both 17alpha-ethinyl substituted oestrogens and progestogens greatly reduced the basal bile flow. The parent compounds, oestradiol-17beta and 19-nortestosterone had little or no effect.3. A much larger dose of progestogens (40 mg/kg) than oestrogens (5 mg/kg) was needed.4. Between 12 and 48 h were required for 17alpha-ethinyloestradiol to produce the effect.5. Bilirubin maximum secretion rate (Tm) was little affected, the only significant reduction being produced by the 3-methyl ether of 17alpha-ethinyloestradiol (mestranol).6. Rises in serum conjugated bilirubin following infusion of bilirubin were produced by 17alpha-ethinyloestradiol and mestranol but not by the progestogens.     

Chronopharmacological characteristics of the action of dehydrocholic acid on bile formation

Peculiarities of the liver response to dehydrocholic acid, conditioned by the time of its administration during the day and by the seasons of the year were revealed in experiments on random bred mature white rats. The dependence of this reaction on the time factor was manifested in all the main indicators of cholopoiesis--in the rate of bile secretion, synthesis and secretion of bile acids and bilirubin, cholesterol excretion.     

芒果苷利胆作用及对胆囊平滑肌痉挛的影响

目的：探讨芒果苷利胆作用及对胆囊平滑肌痉挛的影响.方法：利用胆管引流法测定芒果苷对大鼠胆汁的流量及成分的影响;通过对豚鼠胆囊肌条的收缩试验观察芒果苷对乙酰胆碱所致胆囊平滑肌痉挛的影响.结果：芒果苷4.74×10-5 mol·kg-1剂量组可显著提高大鼠的胆汁流量,显著提高胆汁中胆红素浓度（P ＜0.01）;2.37×10-5 mol·L-1剂量组可显著抑制由乙酰 胆碱引起的豚鼠胆囊痉挛（P＜0.01）.结论：芒果苷具有利胆作用,可缓解乙酰胆碱引起胆囊痉挛.     

Effect of metaphos on the bile-forming function of the liver]

In experiments with albino male rats the effect of a single introduction into the stomach of methaphos in doses of DL50, 1/10 DL50, 1/20 DL50, 1/50 DL50 and also of a daily consumption of the compound for a month was studied. Metaphos was found to exert a manysided action on the cholopoietic function of the liver, viz. it increases the intensity of the bile secretion, inhibits the synthesis of primary bile acids and conjugation of cholic acid with taurine and glycine, stimulates the formation, conjugation and secretion of bilirubin.     

CYCLOSPORINE A HEPATOTOXICITY: EFFECT OF PROLONGED TREATMENT WITH CYCLOSPORINE ON BILIARY LIPID SECRETION IN THE RAT

1. The effects of cyclosporine A (CyA) treatment on liver morphology, bile flow and biliary secretion of bile acid, cholesterol and phospholipid and some plasma biochemical indicators of liver function were examined. 2. Wistar rats were treated i.p. with 10 or 20 mg of CyA/kg per day for 1, 2, 3 or 4 weeks. 3. Treatment increased bile acid and bilirubin plasma concentration. Bile flow and biliary secretion of bile acid, cholesterol and phospholipid were reduced in CyA-treated animals. 4. All these effecs of the drug appeared at 1 week after the start of treatment and were enhanced during prolonged treatment. Cyclosporine A-induced cholestasis was due to a decrease in both the bile acid-dependent and -independent fractions of bile flow. 5. The reduction in cholesterol and phospholipid biliary output may be secondary to the inhibition of the hepatobiliary flux of bile acid; however, perturbations in the removal of lipids from the canalicular membrane as well as intracanalicular interaction between CyA and lipid vesicles/micelles could also be involved.     

Effect of cyclophosphane on bile formation and lipid peroxidation in the liver

In experiments on male albino rats cyclophosphamide (100-250 mg/kg) was shown to produce a hepatic lesion presented with the increase in the activity of alanine- and aspartate aminotransferase of blood serum, and the inhibition of secretion of bile, bile acids and bilirubin. In addition, the drug possesses prooxidant properties: at the doses of 10-250 mg/kg it promotes an increase in the content of malon dialdehyde and diene conjugates in the liver, kidneys, myocardium and lungs. The prooxidant properties of cyclophosphamide were suggested to play an important role in its toxicity.     

Effect of phenobarbital on the bile-forming function of the liver

In tests with 178 albino male-rats the effect of phenobarbital in doses of 30--100 mg/kg, administered intraperinoneally singly and multiply, on the biligenic function of the liver was studied. The drug was found to act beneficially on the intensity of biligenesis, the synthesis of primary bile acids and their cojugation with taurine and glycine, as well as on the formation of and cojugation with glucoronic acid and secretion of bilirubin. This action of phenobarbital continues also in the progeny of the female rats receiving phenobarbital at different periods of pregnancy.     

Tienilic acid enhances hyperbilirubinemia in Eisai hyperbilirubinuria rats through hepatic multidrug resistance-associated protein 3 and heme oxygenase-1 induction.

We demonstrated that tienilic acid, a diuretic drug withdrawn from the market because of hepatic failure, enhanced hyperbilirubinemia in Eisai hyperbilirubinuria rats (EHBR) with a defect of canalicular multidrug resistance-associated protein 2 (Mrp2). In contrast, no remarkable changes were noted in Sprague-Dawley (SD) rats, the parent strain for EHBR. To investigate a mechanism underlying this enhanced hyperbilirubinemia, we focused on comprehensive effects of tienilic acid on clinicopathological aspects and expression of hepatic transporters. Other than eventual hyperbilirubinemia with slightly increased biliary bilirubin, a single oral treatment of EHBR with tienilic acid at 300 mg/kg caused no changes in serum alanine aminotransferase and alkaline phosphatase, bile flow rate and biliary bile acid secretion, or hepatic morphology. In analyses of mRNA expression of the hepatic transporters, elevated Mrp3 expression in EHBR correlated with an increase in serum total bilirubin, suggesting increased bilirubin transport from the liver into the peripheral blood flow. Hepatic heme oxygenase-1 (Ho-1) mRNA, a stress-induced isoform of the rate-limiting enzyme in the catabolism of heme to bilirubin, was markedly upregulated in EHBR at the same dose at which increased serum bilirubin was seen. A time-course study revealed that marked induction of Ho-1 occurred earlier than that of Mrp3, followed by an increase in serum bilirubin. These results suggest that hepatic Mrp3 and Ho-1 may contribute to tienilic acid-enhanced hyperbilirubinemia in EHBR by inducing increased bilirubin transport from the liver into the blood stream, preceded by potentiation of bilirubin formation in the liver.     

Action of phytoecdysteroids on the bile-secretory function of the normal liver and in experimental hepatitis

Phytoecdysteroids from Rhaponticum carthemoides (Willd) Iljin and Ajuga turkestanica (Rgl.) Brig in a dose of 5 mg/kg per os markedly stimulate the bile secretion in normal rats treated with single (cyasterone) and 7-day-long (ecdysterone, cyasterone) administration. The chemical composition of the bile is significantly improved because of increased levels of bile acids and bilirubin and a decreased cholesterol content. Phytoecdysteroids (ecdysterone) exert more beneficial effect on the parameters under study in rats with toxic hepatitis induced by heliotrine.     

Value of bile acid determination for the diagnosis of neonatal jaundice

Serum concentrations of bile acids and bilirubin, and activity of alanine transferase and alkaline phosphatase as well as bile acid and bilirubin levels in duodenal contents were determined in 90 infants aged 1-44 weeks (including 49 under 10 weeks of age) admitted to hospital for prolonged jaundice. Infants with extrahepatic cholestasis were found to have statistically higher serum bile acid and bilirubin concentrations. Oral administration of cholestyramine produced a statistically significant decrease in serum bile acids and bilirubin in infants with intrahepatic cholestasis under 10 weeks of age. In 24 out of the 30 infants with biliary tract obstruction total absence of bile acids in the duodenal contents was demonstrated while in the others the concentration did not exceed 0.2 mmol/l. The mean bile acid concentration in infants with intrahepatic cholestasis was 2.81 mmol/l while in 8 infants out of the 60 bile acids were either absent or present in trace amounts. The method had an 84.4% sensitivity.     

Identification of Urinary Biomarkers Useful for Distinguishing a Difference in Mechanism of Toxicity in Rat Model of Cholestasis

Abstract: This ¹H nuclear magnetic resonance metabonomics study was aimed to determine urinary biomarkers of cholestasis resulting from inhibition of biliary secretion of bile or obstruction of bile flow. To inhibit biliary secretion of bile, cyclosporine A was administered to male Sprague–Dawley rats. Obstruction of bile flow was induced by administration of 4,4′‐methylene dianiline, α‐naphthylisothiocyanate or bile duct ligation. Clinical pathological and histopathological examinations were performed to confirm cholestatic injury and ¹H nuclear magnetic resonance spectral data for urine samples were analysed to determine similarities and differences in profiles of metabolites using the Spotfire^®. In cyclosporine A‐treated groups, serum total bilirubin and bile acid were significantly increased but no remarkable hepatic histopathological‐changes were observed. In 4,4′‐methylene dianiline‐, α‐naphthylisothiocyanate‐ and bile duct ligation‐treated groups, serum alkaline phosphatase, γ‐glutamyltranspeptidase and total bilirubin levels increased significantly, and hepatic histopathological‐changes were observed. On urinary ¹H nuclear magnetic resonance spectral analysis, area intensities derived from 0.66 to 1.90 ppm were decreased by cyclosporine A, whereas they were increased by other treatments. These metabolites were identified using the NMR suite^® as bile acids, branched‐chain amino acids, n‐butyrate, propionate, methyl malonate and valerate. These metabolites were further investigated by K‐means clustering analysis. The cluster of these metabolites is considered to be altered by cholestasis. We conclude that bile acids, valine and methyl malonate have a possibility to be urinary cholestatic biomarkers, which distinguish a difference in mechanism of toxicity. ¹H nuclear magnetic resonance metabonomics thus appears to be useful for determining the mechanisms of toxicity and can be front‐loaded in drug safety evaluation and biomarker discovery.     

The comparative action of isoniazid, rifampicin and ethambutol on liver function]

It has been established in experiments on white rats that antituberculous drugs (isoniazid, rifampicin, ethambutol) given in toxic doses affect the liver, its membranes and organelles, inhibit bile production and bioenergy. This is supported by activation of aspartate and alanine aminotransferases, (ALT and AST), alkaline phosphatase in blood serum and acid phosphatase in the liver, by a decrease of the activity of Na(+)-, K(+)-ATPase, succinate dehydrogenase and cytochromoxidase in the liver, lowering of the rate of bile secretion, excretion of bile acids, bilirubin and cholesterol with bile. Provided the drugs are administered in combination, the hepatotoxicity rises, particularly in combination of isoniazid with rifampicin, and especially as isoniazid is combined with rifampicin and ethambutol.     

虎杖水提液利胆保肝作用研究

目的初步研究虎杖水提液的利胆保肝作用。方法以大鼠胆管插管法,十二指肠给药,记录给药前1h,给药后1,2,3和4h胆汁流量及胆汁中胆红素(TBIL)和胆固醇(CHO)含量,观察虎杖水提液利胆作用;采用腹腔注射CCl4橄榄油溶液复制小鼠急性肝损伤模型,检测血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)含量,观察其对肝的保护作用。结果虎杖水提液可增加大鼠胆汁分泌量(P<0.05或P<0.01);可明显降低CCl4模型小鼠血清ALT和AST含量(P<0.05或P<0.01)。结论虎杖水提液具有利胆和保肝作用。     

Antioxidant effectiveness in isoniazid-induced lesions of the liver

In male albino rats tocopherol acetate and sodium selenite were shown to be efficient in treatment of isoniazide-induced liver damage. The disturbances of secretion of bile, bile acids and bilirubin and excretion of cholesterol were less pronounced. A decrease of the activity of marker blood serum enzymes--alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase--also indicated the hepatoprotective effect of these drugs. The intensity of lipoperoxidation indicated by a decrease of lipid peroxidation indices in the liver and blood was diminished. Tocopherol acetate and sodium selenite increase the number of sulfhydryl groups but decrease the number of disulfide groups in these biosubstrates.     

The effect of galbanic acid on the course of experimental hepatitis

Administration of galbanic acid isolated from the roots of Ferula kopetdaghensis Eug. Kor. to rats orally in a dose of 50 mg/kg was found to improve the course of toxic hepatitis induced by fourfold subcutaneous injections of a 50% oil solution of CCl4. Galbanic acid produced much more earlier than in control normalization of the activity of the enzymes alanine- and aspartataminotransferase in blood serum, increased glycogen content and improved the parameters of the redox potential of lactic acid-pyruvic acid system in the liver. Galbanic acid exerted the antioxidant effect. In the animals with a developed hepatitis receiving galbanic acid there was a more rapid restoration of intensity of bile secretion, synthesis of bile acids and bilirubin, cholesterol excretion as compared with control.     

舒胆通颗粒对实验动物胆汁分泌及胆囊平滑肌的影响

目的观察舒胆通（SDT）颗粒对大鼠胆汁分泌及胆囊平滑肌的影响。方法采用在体胆汁引流法,观察单次给予SDT对大鼠胆汁分泌以及总胆红素（TB）的影响;采用离体试验法观察SDT对豚鼠胆囊平滑肌条的直接作用及其对阿托品、酚妥拉明、苯海拉明、雷尼替丁的拮抗作用。结果SDT 5,10,20 g.kg^-1剂量能使大鼠给药后30－90 min时段胆汁分泌增加,但胆汁中TB水平无明显改变;终浓度为2.5×10^-4,5×10^-4,1×10^-3 mg.mL^-1的SDT能使离体豚鼠胆囊肌条张力增高、收缩频率加快、收缩幅度增加,并呈现明显的量效关系,其兴奋作用可被阿托品完全阻断,也可被酚妥拉明部分阻断,但不能被苯海拉明、雷尼替丁所阻断。结论SDT能增强胆囊平滑肌的兴奋性,促进胆汁分泌;其利胆作用可能主要经由胆碱能M受体介导,部分经由肾上腺素能α受体介导、而与组胺H1受体、前列腺素受体等途径无关。