Artemisia capillaris Thunb. Polysaccharide alleviates cholestatic liver injury through gut microbiota modulation and Nrf2 signaling pathway activation in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: According to traditional Chinese medicine (TCM) theory, cholestasis belongs to category of jaundice. Artemisia capillaris Thunb. has been widely used for the treatment of jaundice in TCM. The polysaccharides are the one of main active components of the herb, but its effects on cholestasis remain unclear. AIM OF THE STUDY: To investigate the protective effect and mechanism of Artemisia capillaris Thunb. polysaccharide (APS) on cholestasis and liver injury. MATERIALS AND METHODS: The amelioration of APS on cholestasis was evaluated in an alpha-naphthyl isothiocyanate (ANIT)-induced mice model. Then nuclear Nrf2 knockout mice, mass spectrometry, 16s rDNA sequencing, metabolomics, and molecular biotechnology methods were used to elucidate the associated mechanisms of APS against cholestatic liver injury. RESULTS: Treatment with low and high doses of APS markedly decreased cholestatic liver injury of mice. Mechanistically, APS promoted nuclear translocation of hepatic nuclear factor erythroid 2-related factor (Nrf2), upregulated downstream bile acid (BA) efflux transporters and detoxifying enzymes expression, improved BA homeostasis, and attenuated oxidative liver injury; however, these effects were annulled in Nrf2 knock-out mice. Furthermore, APS ameliorated the microbiota dysbiosis of cholestatic mice and selectively increased short-chain fatty acid (SCFA)-producing bacteria growth. Fecal microbiota transplantation of APS also promoted hepatic Nrf2 activation, increased BA efflux transporters and detoxifying enzymes expression, ameliorated intrahepatic BA accumulation and cholestatic liver injury. Non-targeted metabolomics and in vitro microbiota culture confirmed that APS significantly increased the production of a microbiota-derived SCFA (butyric acid), which is also able to upregulate Nrf2 expression. CONCLUSIONS: These findings indicate that APS can ameliorate cholestasis by modulating gut microbiota and activating the Nrf2 pathway, representing a novel therapeutic approach for cholestatic liver disease.

Therapeutic effect of yinchenhao decoction on cholelithiasis via mucin in the gallbladder and intestine.

Cholelithiasis is a common and frequently occurring disease worldwide that belongs to the category of jaundice in traditional Chinese medicine. Yinchenhao decoction (YD) consists of Artemisia capillaris Thunb., Gardenia jasminoides J.Ellis, and Rheum palmatum L., and is traditionally used to treat jaundice, which has a significant therapeutic effect on cholelithiasis. Our study aimed to investigate the pathological mechanism of cholelithiasis and the therapeutic mechanism of YD via mucin in the gallbladder and intestine. YD was prepared and analyzed using HPLC. The supersaturation stability experiment was designed by the solvent-shift method. The cell transport experiment was conducted by coculture monolayers. The animal experiment was performed using a cholelithiasis model with a high-cholesterol diet. The related indicators were detected by automatic biochemical analyzer, PCR, western blot, or ELISA. Statistics were analyzed using χ2-tests and t-tests. As the results, in cholelithiasis, MUC5AC highly expressed in the gallbladder shortened cholesterol supersaturation and promoted cholesterol crystallization via the inflammatory cytokine signaling pathway; MUC2 highly expressed in the small intestine prolonged cholesterol supersaturation and promoted cholesterol absorption via the inflammatory cytokine signaling pathway. YD inhibited mucin expression in the gallbladder and intestine in a concentration-dependent manner for cholelithiasis treatment by inhibiting the inflammatory cytokine signaling pathway, which was attributed to the active components, including chlorogenic acid, geniposide, and rhein.

Daily administration times of canhuang tablet based on a pharmacodynamic/pharmacokinetic model in jaundiced rats.

OBJECTIVE: To study the daily administration times of Canhuang tablet (CHT) for treating jaundice in rats based on a pharmacodynamic/pharmacokinetic model. METHODS: Rats were modeled by 4% 1-Naphthylisothiocyanate (75 mg/kg, p.o.). After 48 h, CHT was given (p.o.) at 0.75 g/kg once a day, 0.375 g/kg twice a day, and 0.25 g/kg three times a day. Blood was collected from the orbital sinus at different intervals. Levels of liver enzymes and bilirubin were detected using these blood samples. Bile was collected and determined after the first administration of CHT. High-performance liquid chromatography was used to determine the concentration of berberine in bile simultaneously. Time-effect and time-dose curves were then obtained. RESULTS: Compared with rats taking CHT twice and three times a day, the total amount of bile within 10 h of rats taking CHT once a day were 1.32- and 1.47-fold higher, respectively. There was good consistency between the pharmacokinetics of berberine and the pharmacodynamics of the effect on liver enzymes and bilirubin in vivo. The pharmacokinetic analyses showed that rats administered CHT once daily maintained a higher concentration of berberine in bile for a longer period than rats administered CHT two- and three-times daily. CONCLUSION: In jaundiced rats, taking CHT once a day is better than taking CHT twice or three times a day. These data may provide a reference for the clinical application of CHT.

Beyond plasma bilirubin: the effects of phototherapy and albumin on brain bilirubin levels in Gunn rats.

BACKGROUND & AIMS: Severe unconjugated hyperbilirubinemia, as occurs in Crigler-Najjar disease and neonatal jaundice, carries the risk of neurotoxicity. This neurotoxicity is related to the increased passage of free bilirubin (UCB(free)), the fraction of bilirubin that is not bound to plasma proteins, into the brain. We hypothesized that albumin treatment would lower the UCB(free) fraction, and thus decrease bilirubin accumulation in the brain. METHODS: We treated chronic (e.g., as a model for Crigler-Najjar disease) and acute hemolytic (e.g., as a model for neonatal jaundice) moderate hyperbilirubinemic Gunn rats with phototherapy, human serum albumin (HSA) or phototherapy+HSA. RESULTS: In the chronic model, adjunct HSA increased the efficacy of phototherapy; it decreased plasma UCB(free) and brain bilirubin by 88% and 67%, respectively (p<0.001). In the acute model, adjunct HSA also increased the efficacy of phototherapy; it decreased plasma UCB(free) by 76% (p<0.001) and completely prevented the hemolysis-induced deposition of bilirubin in the brain. Phototherapy alone failed to prevent the deposition of bilirubin in the brain during acute hemolytic jaundice. CONCLUSIONS: We showed that adjunct HSA treatment decreases brain bilirubin levels in phototherapy-treated Gunn rats. We hypothesize that HSA decreases these levels by lowering UCB(free) in the plasma. Our results support the feasibility of adjunct albumin treatment in patients with Crigler-Najjar disease or neonatal jaundice.

Therapeutic effect of traditional Chinese medicine on coagulation disorder and accompanying intractable jaundice in hepatitis B virus-related liver cirrhosis patients.

AIM: To observe the therapeutic effects of new traditional Chinese medicine (TCM) therapy on coagulation disorder and accompanying intractable jaundice in HBV-related liver cirrhosis patients. METHODS: Using stratified random sampling according to fibrinogen (Fib) levels, 145 liver cirrhosis patients due to hepatitis B complicated by coagulation disorder were treated. Of them, 70 in research group were treated with TCM by "nourishing yin, cooling blood and invigorating blood circulation" and Western medicine, 75 in control group were treated with conventional Western medicine. The indexes of liver function, coagulation function and bleeding events were observed and compared. RESULTS: The prothrombin time (PT) was shorter and the fibrinogen (Fib) level was higher in the research group than in the control group (Fib = 1.6-2.0 g/L, 1.1-1.5 g/L, and < or = 1.0 g/L). The total bilirubin (TBIL) level was significantly lower in the research group than in the control group, except for the subgroup of FIB < or = 1.0 g/L. CONCLUSION: TCM therapy can improve coagulation fuction and decrease TBIL.

Liver injury induced by a Japanese herbal medicine, sairei-to (TJ-114, Bupleurum and Hoelen Combination, Chai-Ling-Tang) R1.

The case is reported of a man who showed acute hepatitis with jaundice after he was given a Japanese herbal medicine, sairei-to (TJ-114, Bupleurum and Hoelen Combination, Chai-Ling-Tang). Unusually, the component thought to be responsible for the observed drug-induced liver injury was able to be identified. Lymphocyte migration inhibition testing indicated that the tuber of the perennial herbage Pinellia ternate was the causative agent.

[Icterus of the newborn caused by indirect bilirubin--recent progress]. / Ujszülöttek indirekt bilirubin okozta icterusa--újabb ismeretek és szempontok.

Recently a big shift has taken place in the judgment and treatment of jaundice in newborn, caused by increased unconjugated bilirubin level. New techniques evolved for assessing the prognosis of developing jaundice. An important major discovery is the antioxidant effect of bilirubin. We have a broader range of knowledge concerning the mechanism of bilirubin toxicity and for judging the chance of developing kernicterus. The prevention techniques do not stop at prohibiting anti-D immunisation but go on to preventing hydrops foetalis, the life-threatening form of haemolytic disease. There are data about the complications of phototherapy and EPO treatment for prolonged anaemia.

[Hepatocyte apoptosis and the expression of Bcl-2 and Bax in Yin-jaundice rats].

OBJECTIVE: To investigate the effects of Yinchen Shufu Decoction on hepatocyte apoptosis and the expression of its apoptosis-regulating gene Bcl-2 and Bax in Yin-jaundice rats. METHODS: Forty-eight rats were divided randomly into 4 groups: the normal control group, Yin-jaundice model group, Yang-jaundice model group, Yinchen Shufu Decoction treatment group. The TUNEL assay and the immunohistochemistry assay were used to detect the apoptosis of hepatocytes and the expression of Bcl-2 and Bax in hepatocytes respectively. RESULTS: The rate of apoptosis cells in Yin-jaundice model group was higher significantly than that in Yang-jaundice model group and normal control group (P<0.01), the expression of Bcl-2 in Yinchen Shufu Decoction treatment group was higher significantly than that in Yin-jaundice model group (P<0.01 or P<0.05), and the expression of Bax in it was lower significantly than that in Ying-jaundice model group (P<0.01 or P<0.05). CONCLUSION: Yinchen Shufu Decoction can prevent hepatocyte apoptosis perhaps by up-regulating the expression of Bcl-2 and down-regulating the expression of Bax. It is one of the mechanisms of its treatment on Yin-jaundice.

An oral endothelin-A receptor antagonist blocks collagen synthesis and deposition in advanced rat liver fibrosis.

BACKGROUND & AIMS: Endothelin 1 induces contraction, proliferation, and collagen synthesis of hepatic stellate cells in vitro, which may be mediated via the endothelin A receptor. It is unknown if specific blockade of the endothelin A receptor inhibits hepatic fibrosis in vivo. METHODS: Groups of 10-20 rats with bile duct occlusion were treated with the nonpeptide endothelin-A receptor antagonist LU 135252 at 80 mg. kg(-1). day(-1) from week 1-6 or from week 4-6, or with LU at 10 mg. kg(-1). day(-1) from week 1-6. Animals with bile duct occlusion alone and sham-operated rats without or with LU at 80 mg. kg(-1). day(-1) over 6 weeks served as controls. After 6 weeks, parameters of fibrogenesis were determined. RESULTS: LU treatment led to improved histology, paralleled by a dose-dependence up to 60% reduction of liver collagen, even when administered at an advanced fibrosis stage. This was accompanied by a decreased messenger RNA of hepatic procollagen alpha1(I) and tissue inhibitor of metalloproteinase 1, 2 major effectors of fibrosis, and of serum procollagen type III, a surrogate marker of liver fibrogenesis. CONCLUSIONS: Selective endothelin-A receptor blockade can dramatically reduce collagen accumulation in rat secondary biliary fibrosis, a model refractory to most potential antifibrotic agents. Endothelin-A receptor antagonists are promising antifibrotic agents in chronic liver disease.

Vitamin A status in biliary atresia: intestinal absorption and liver storage of retinol.

The vitamin A status of 19 patients with corrected biliary atresia was examined. They had been receiving 5,000 IU of oral vitamin A daily postoperatively. Plasma vitamin A levels in the nonjaundiced group were almost within normal range, whereas those in the jaundiced group were significantly low compared with the controls. In the oral vitamin A tolerance test, plasma vitamin A levels increased from 33.1 +/- 11.8 to 215.4 +/- 100.7 micrograms/dL in the nonjaundiced group, and from 23.1 +/- 10.3 to 209.8 +/- 154.2 micrograms/dL in the slightly jaundiced group, at 4 hours after the administration of vitamin A, showing no difference between both group and control. In the severely jaundiced group, plasma vitamin A levels increased from 13.5 +/- 3.5 to 30.0 +/- 14.6 micrograms/dL, a significantly smaller increase compared with controls. However, liver vitamin A levels were greater than 20 micrograms/g liver in all patients, irrespective of the presence of jaundice. This study suggested that nutritional support to facilitate the synthesis of retinol-binding protein may be an important factor in addition to vitamin A supplementation.

Four-dimensional chemical shift MR imaging of phosphorus metabolites of normal and diseased human liver.

Four dimensional chemical shift imaging was used to map the relative peak heights of phosphorus metabolites of the liver and overlying skeletal muscle of a normal subject and two patients. The technique provides 31P spectra localised on a voxel-by-voxel basis and may be valuable in mapping heterogeneous structural and metabolic changes in disease.

Unconjugated bilirubin and the bile from light exposed Gunn rats inhibit intestinal water and electrolyte absorption.

Jaundiced babies undergoing phototherapy often develop diarrhoea. The cause of it is still uncertain. Increasing evidence supports a role of a secretory mechanism for the diarrhoea. We therefore studied the effects of bile from congenitally jaundiced rats undergoing phototherapy and of unconjugated bilirubin on rat small intestine in vivo and in vitro. Results suggest that: (1) the bile from homozygous Gunn rats under phototherapy has an anti-absorptive effect when tested in the perfused jejunum of normal Wistar rats; (2) unconjugated bilirubin has a dose dependent secretory effect on the intestinal transport of water and electrolytes, when tested in the same system. Alteration of cyclic AMP or cyclic GMP, known intracellular mediators of secretion, was not observed. We conclude that free bilirubin is an intestinal secretagogue acting by an as yet unknown mechanism, that may mediate the secretory type of diarrhoea in jaundiced neonates undergoing phototherapy.

Effect of combined treatment with 2,3,7,8-tetrachlorodibenzo-p-dioxin and phototherapy on bilirubin metabolism in the jaundiced Gunn rat.

2,3,7,8-Tetrachlorodibenzo-p-dioxin, a potent inducer of microsomal cytochrome P448-dependent monoxygenases, and phototherapy both accelerate bilirubin metabolism and decrease jaundice in Gunn rats. The effects of combined treatment with 2,3,7,8-tetrachlorodibenzo-p-dioxin and light were studied in these rats by applying phototherapy for 65 hr, beginning 5 days after induction with 2,3,7,8-tetrachlorodibenzo-p-dioxin. 2,3,7,8-Tetrachlorodibenzo-p-dioxin pretreatment caused a 75% decline in plasma bilirubin in 5 days, with no change thereafter, whether or not the rats were exposed subsequently to phototherapy. In the uninduced rats, plasma bilirubin levels declined by 55% after 40 hr of phototherapy. As determined by [14C]bilirubin kinetics, both 2,3,7,8-tetrachlorodibenzo-p-dioxin and phototherapy increased fractional bilirubin turnover and decreased the total bilirubin pool. In the 2,3,7,8-tetrachlorodibenzo-p-dioxin-induced rats, the contracted bilirubin pool shifted from skin to liver, but these tissue pools did not change further during phototherapy. By contrast, in uninduced rats, phototherapy decreased the cutaneous bilirubin pool, which is the main target of phototherapy. 2,3,7,8-Tetrachlorodibenzo-p-dioxin was more effective than phototherapy in diminishing plasma bilirubin levels and the total bilirubin pool, but the combined treatment (2,3,7,8-tetrachlorodibenzo-p-dioxin followed by phototherapy) was no more effective than 2,3,7,8-tetrachlorodibenzo-p-dioxin alone.

Effects of phototherapy on the activity of microsomal mixed-function monooxygenases and the pharmacokinetics of [14C] hexobarbital in jaundiced and heterozygous Gunn rats.

We assessed the effects of in vivo phototherapy of Gunn rats on the activity of hepatic microsomal mixed-function monoxygenases and on the in vivo pharmacokinetics of [14C]HB. In experiment 1 no serial changes were seen in activities of hexobarbital hydroxylase or benzo(a)pyrene hydroxylase in hepatic microsomes isolated after 2, 4 or 7 days from homozygous jaundiced female Gunn rats exposed to continuous phototherapy or in matched Gunn rats maintained under dim light. In experiment 2 homozygous jaundiced (jj) and heterozygous nonjaundiced (Jj) Gunn rats of both sexes each received i.v. [14C]HB on 2 successive days. In random order, each was exposed on the first or second day to phototherapy for 5.5 hr, beginning 0.5 hr before the administration of HB; otherwise, each was kept under dim light. Plasma [14C]HB in arterial blood samples was separated chromatographically from its labeled metabolites, and biexponential plasma disappearance curves for [14C]HB were analyzed by a SAAM-23 computer program. Clearances in female rats were much slower. In both sexes, the total body clearance and volume of distribution of HB were decreased by 20% during phototherapy of the jj but not the Jj rats; terminal plasma half-life was unchanged. In experiment 3 direct in vitro illumination of [14C]HB did not cause photodegradation of this compound, despite the presence of albumin with or without bilirubin.

Selenium concentration in maternal and umbilical cord blood, placenta and amniotic membranes.

The selenium concentration of maternal and umbilical cord whole blood was determined by atomic absorption spectrophotometry in 21 parturients at term. Six placental and amniotic membrane tissue specimens were also investigated. The mean selenium concentrations in the maternal (0.73 +/- 0.15 mumol/l) and umbilical cord blood (0.77 +/- 0.18 mumol/l) were similar and without significant correlation. Placental (2.24 +/- 0.20 mumol/kg wet weight) and amniotic membrane tissue specimens (2.32 +/- 0.54 mumol/kg wet weight) also contained similar concentrations of selenium which were about 3 times higher than those in the maternal and umbilical cord blood. Low whole blood selenium concentration in Finnish parturients may be a sign of deficient nutritional intake of selenium during pregnancy. The relatively high concentration of selenium in the placenta and amniotic membranes on the other hand suggest that metabolically active organs are being provided primarily with this essential trace element.

The effect of elimination of environmental light on the metabolism of unconjugated bilirubin in the Gunn rat.

In the homozygous jaundiced Gunn rat, bilirubin catabolism is augmented by intense illumination (phototherapy) and by induction of microsomal cytochrome P448. To assess the relative importance of less intense environmental light versus intrinsic mechanisms in the maintenance of bilirubin turnover, Gunn rats were kept for three weeks under either ordinary laboratory lighting (0.3-0.8 mW/cm2, wavelength range 400-600 nm) or in absolute darkness. No differences in plasma concentration, miscible pool, turnover of bilirubin, or in hepatic cytochrome P448 activity were noted between the two groups over this period. A greater than twofold increase in the biliary excretion of unconjugated bilirubin was noted in the animals maintained under light, but this represented only 2% of total bilirubin turnover. These results suggest that intrinsic(enzymatic ?) pathways are of primary importance in the maintenance of bilirubin metabolism in the glucuronyltransferase-deficient state under ordinary levels of environmental light.

Characteristic changes of cerebellar proteins associated with cerebellar hypoplasia in jaundiced Gunn rats and the prevention of these by phototherapy.

In the cerebellar particulate fractions from Gunn rat homozygotes 3 protein bands with apparent mol. wts. of 250,000, 50,000 and 33,000 in SDS-polyacrylamide gel disc electrophoresis underwent major changes, and phototherapy of the newborns could effectively prevent changes.