Systems Toxicology Approaches Reveal the Mechanisms of Hepatotoxicity Induced by Diosbulbin B in Male Mice.

Diosbulbin B (DIOB) is an effective component of air potato yam with antitumor and anti-inflammatory activities, and it is the main toxic component leading to hepatotoxicity. However, the mechanism of its hepatotoxicity remains unclear. In this study, we aimed to systematically elucidate the molecular action of DIOB on liver metabolic function through systems toxicology approaches. C57BL/6 mice were orally treated with DIOB (10, 30, 60 mg/kg) for 28 days, and the liver metabonomics and histopathology, molecular docking, mRNA expression levels, and activities of enzymes were analyzed. The results illustrated that DIOB could affect fatty acid and glucose metabolism, block the TCA cycle, and DIOB also could disorder bile acid synthesis and transport and promote the occurrence of hyperbilirubinemia. In addition, DIOB increased Cyp3a11 expression in a dose-dependent manner. Thus, these results provide new insights into the mechanism of hepatotoxicity caused by DIOB.

Untargeted LC-MS-based metabonomics revealed that aristolochic acid I induces testicular toxicity by inhibiting amino acids metabolism, glucose metabolism, ß-oxidation of fatty acids and the TCA cycle in male mice.

As the main toxic component of aristolochic acid, aristolochic acid I (AAI) is primarily found in Aristolochiaceae plants such as Aristolochia, Aristolochia fangchi and Caulis aristolochiae manshuriensis. AAI has been proven to be carcinogenic, mutagenic and nephrotoxic. Although the role of AAI in testicular toxicity has been reported, its mechanism of action is unknown. Using metabonomics and molecular biology techniques, we tried to identify the differential endogenous metabolites of AAI that may affect the changes in testicular function in mice, map the network of metabolic pathways, and systematically reveal the molecular mechanism of AAI-induced testicular toxicity. We found that AAI inhibited amino acid metabolism in mouse testicular cells, impeded the uptake and oxidative decomposition of fatty acids, prevented normal glucose uptake by testicular cells, which inhibited glycolysis and gluconeogenesis, affected the mitochondrial tricarboxylic acid (TCA) cycle, which impaired the ATP energy supply, decreased the number of spermatogenic cells and sperm in the testes, induced changes in the mitochondrial state of spermatogonial cells, and ultimately led to physiological and pathological changes in the testes. AAI also regulated the testicular physiological activity by regulating the androgen receptor and hormone levels. This study used metabonomics and other methods to elucidate the mechanism of AAI-induced testicular toxicity from a new angle.

Quadrupole Orbitrap Mass Spectrometer-Based Metabonomic Elucidation of Influences of Short-Term Di(2-ethylhexyl) phthalate Exposure on Cardiac Metabolism in Male Mice.

Di(2-ethylhexyl) phthalate (DEHP) can cause severe environmental pollution. Effects of DEHP on cardiac metabolism have been reported, but its mechanism(s) of action is not fully clear. Here, we used high-resolution mass spectrometry for metabonomics and molecular biological methods to identify the different endogenous metabolites affected by DEHP that might cause changes in cardiac metabolism in mice, map the network of metabolic pathways, and reveal (at the molecular level) how DEHP affects cardiac metabolism. The results showed that DEHP could inhibit the ß-oxidation of fatty acids and gluconeogenesis, promote glycolysis, and inhibit the tricarboxylic acid cycle in cardiomyocytes. DEHP caused mitochondrial dysfunction by inhibiting the synthesis and transport of fatty acids and, thus, inhibiting the synthesis and breakdown of adenosine triphosphate in mitochondria. Pathology revealed that DEHP could change the normal structures and functions of the heart and bodies of mice. DEHP can interfere with the physiological and metabolic function of the heart in mice by disrupting the endogenous metabolite and gene levels.

Simultaneous quantification of L-tetrahydropalmatine and its urine metabolites by ultra high performance liquid chromatography with tandem mass spectrometry.

l-tetrahydropalmatine (l-THP) is a tetrahydroprotoberberine isoquinoline alkaloid that has been used as an analgesic agent in China for more than 40 years. Recent studies indicated its potential application in the treatment of drug addiction. In this study, a sensitive and rapid method using ultra high performance liquid chromatography with MS/MS was developed and validated for simultaneous quantitation of l-THP and its desmethyl metabolites. Enzymatic hydrolysis was integrated into sample preparation to enable the quantitative determination of both free and conjugated metabolites. Chromatographic separation was achieved on an Agilent Poroshell 120 EC-C18 column. Detection was performed by MS in the positive ion ESI mode. The calibration curves of the analytes were linear (r(2) > 0.9936) over the concentration range of 1-1000 ng/mL with the lower limit of quantification at 1 ng/mL. The precision for both intra- and interday determinations was <8.97%, and the accuracy ranged from -8.74 to 8.65%. The recovery for all the analytes was >70% without significant matrix effect. The method has been successfully applied to the urinary excretion study of l-THP in rats. The conjugates were found to be the major urine metabolites of the drug.

Self-other differences in H1N1 flu risk perception in a global context: a comparative study between the United States and China.

Extending research on self-other differences in perception to a global health risk, this study compares U.S. and Chinese college students' perceived H1N1 flu risk at four levels: personal, group, societal, and global. It also examines how personal experience, interpersonal communication, traditional and Internet-based media, and self-efficacy affect perception at four levels, as well as the self-other differences between the personal level and each of the other three levels. An online survey in both countries reveals an "ascending pattern," showing higher perceived risk for others than for selves. Chinese respondents perceive higher risk than U.S. respondents at all levels. Interpersonal communication predicts risk perception at four levels in the United States and at the group and societal levels in China. New media exposure exerts influence on all but the group level in China, while social networking sites (SNS) exposure predicts group- and societal-level risk perception in the United States. The overall attention paid to H1N1 information in the media affects all levels in both countries. Interaction between media exposure and attention is influential at all levels in the United States. Self-efficacy is negatively associated with risk perception in China except at the global level. Attention to media in the United States, and SNS exposure in China, explain the self-other differences in three comparisons, along with self-efficacy, which decreases the self-other gap in the United States while increasing the gap in China.

[Investigation of metabolic kinetics and reaction phenotyping of ligustrazin by using liver microsomes and recombinant human enzymes].

The metabolic characteristics of ligustrazin (TMPz) in liver microsomes were investigated in the present study. The reaction phenotyping of TMPz metabolism was also identified by in vitro assessment using recombinant human cytochrome P450 enzymes (CYP) and UDP glucuronosyltransferases (UGT). TMPz was incubated at 37 degrees C with human (HLM) and rat liver microsomes (RLM) in the presence of different co-factors. The metabolic stability and enzyme kinetics of TMPz were studied by determining its remaining concentrations with a LC-MS/MS method. TMPz was only metabolically eliminated in the microsomes with NADPH or NADPH+UDPGA. In the HLM and RLM with NADPH+UDPGA, t1/2, K(m) and V(max) of TMPz were 94.24 +/- 4.53 and 105.07 +/- 9.44 min, 22.74 +/- 1.89 and 33.09 +/- 2.74 micromol x L(-1), 253.50 +/- 10.06 and 190.40 +/- 8.35 nmol x min(-1) x mg(-1) (protein), respectively. TMPz showed a slightly higher metabolic rate in HLM than that in RLM. Its primary oxidative metabolites, 2-hydroxymethyl-3, 5, 6-trimethylpyrazine (HTMP), could undergo glucuronide conjugation. The CYP reaction phenotyping of TMPz metabolism was identified using a panel of recombinant CYP isoforms (rCYP) and specific CYP inhibitors in HLM. CYP1A2, 2C9 and 3A4 were found to be the major CYP isoforms involved in TMPz metabolism. Their individual contributions were assessed b) using the method of the total normalized rate to be 19.32%, 27.79% and 52.90%, respectively. It was observed that these CYP isoforms mediated the formation of HTMP in rCYP incubation. The UGT reaction phenotyping of HTMP glucuronidation was also investigated preliminarily by using a panel of 6 UGT isoforms (rUGT). UGT1A1, 1A4 and 1A6 were the predominant isoforms mediated the HTMP glucuronidation. The results above indicate that the metabolism of TMPz involves multiple enzymes mediated phase I and phase II reactions.

Phenylalanine deprivation inhibits multiple myeloma progression by perturbing endoplasmic reticulum homeostasis.

Amino acid metabolic remodeling is a hallmark of cancer, driving an increased nutritional demand for amino acids. Amino acids are pivotal for energetic regulation, biosynthetic support, and homeostatic maintenance to stimulate cancer progression. However, the role of phenylalanine in multiple myeloma (MM) remains unknown. Here, we demonstrate that phenylalanine levels in MM patients are decreased in plasma but elevated in bone marrow (BM) cells. After the treatment, phenylalanine levels increase in plasma and decrease in BM. This suggests that changes in phenylalanine have diagnostic value and that phenylalanine in the BM microenvironment is an essential source of nutrients for MM progression. The requirement for phenylalanine by MM cells exhibits a similar pattern. Inhibiting phenylalanine utilization suppresses MM cell growth and provides a synergistic effect with Bortezomib (BTZ) treatment in vitro and murine models. Mechanistically, phenylalanine deprivation induces excessive endoplasmic reticulum stress and leads to MM cell apoptosis through the ATF3-CHOP-DR5 pathway. Interference with ATF3 significantly affects phenylalanine deprivation therapy. In conclusion, we have identified phenylalanine metabolism as a characteristic feature of MM metabolic remodeling. Phenylalanine is necessary for MM proliferation, and its aberrant demand highlights the importance of low-phenylalanine diets as an adjuvant treatment for MM.

Assessment of the roles of P-glycoprotein and cytochrome P450 in triptolide-induced liver toxicity in sandwich-cultured rat hepatocyte model.

Triptolide (TP), a main bioactive component of Tripterygium wilfordii Hook F., is a promising agent for treatment of autoimmune diseases. However, a high incidence of dose-limiting hepatotoxicity was observed in the clinic. Sandwich-cultured rat hepatocyte model was used in this study to identify the involvement of P-glycoprotein (P-gp) in TP disposition and to evaluate TP-induced hepatotoxicity after modulation of P-gp by the known inhibitors, ritonavir and tariquidar, and known inducers, phenobarbital, quercetin, and H(2)O(2). Our data showed that biliary clearance of TP reduced 73.7% and 84.2% upon treatment of ritonavir (25 µM) and tariquidar (5 µM), respectively. In contrast, increases of 346%, 280%, and 273% in biliary clearance of TP were observed with treatment of phenobarbital (1.0 mM), quercetin (20 µM), and H(2)O(2) (0.5 mM), respectively. The TP-induced hepatotoxicity increased by twofold when CYP activity was blocked by 1-aminobenzotriazole, suggesting that CYP and P-gp may both contribute to the detoxification of TP in the SCRH model. In addition, hepatotoxicity and the expression of apoptosis proteins Bax and Bcl-2 were correlated qualitatively with the TP exposure duration and its intracellular concentration, which, in turn, can be modulated by P-gp inhibitors or inducers. Our results for the first time demonstrated that in addition to CYP-mediated metabolism, P-gp also plays an important role in the disposition of TP and TP-induced hepatotoxicity. Thus, the modulation of canalicular P-gp has a potential to cause drug-drug interaction between TP and the coadministered P-gp inhibitors or inducers in the clinic.

[Effect of shenfu injection on CYP450s of rat liver].

The paper is to report the study of the effect of Shenfu injection on the enzyme activity of liver CYP450 and its mRNA level of rat liver. Microsome of rat liver was prepared after intravenous administration of Shenfu injection for 7 days. The enzyme activity was quantified by Cocktail method. Meanwhile, the mRNA expression of CYP1A2, CYP2B1/2, CYP2C11 and CYP3A1 in the liver was detected by RT-PCR. Shenfu injection obviously induced the enzyme activities of CYP2B and CYP2C. Meantime Shenfu injection decreased the enzyme activities of CYP1A2 and CYP3A. The mRNA levels of CYP2B and CYP2C were also induced in rats treated with Shenfu injection. But it obviously inhibited the mRNA level of CYP1A2 and CYP3A. Since the enzyme activity and mRNA level were obviously changed after administration, the potential effect of drug-drug interaction should be concerned.

[Effect of Siwu decoction and its combined administration on hepatic P450 enzymatic activity and mRNA expression in rats].

To study the effect of Siwu decoction (SWD) compound and its combined administration on hepatic P450 enzymatic activity and mRNA expression in rats. Rats were orally administered with SWD and water decoction combined with other medicines for two weeks, and then sacrificed. Their livers were perfused with normal saline to prepare liver micrisomes. Mixed probe and liver microsome in vitro incubation method were adopted to detect the effect of SWD on hepatic cytochrome P450. The real-time quantitative polymerase chain reaction (Q-PCR) was used to detect the effect of SWD on the expression of hepatic cytochrome P450. Compared with the control group, the SWD compound group showed higher CYP1A2 enzymatic activity (P < 0.05); Rehmanniae-paeoniae, angelicae-paeoniae, angelicae-rhizome, paeoniae-rhizome groups had lower CYP1A2 and CYP2C19 enzymatic activities (P < 0.05); And the compound group, the single component group and the combination group showed lower CYP2B6 enzymatic activities (P < 0.05). The compound could up-regulated the mRNA expression of CYP2B1 (P < 0.05); And the four single components could down-regulated the mRNA expression of CYP2B1 (P < 0.05). SWD compound had the effect in inducing CYP1A2 enzymatic activity. The rehmanniae-paeoniae group and the angelicae-paeoniae group had identical enzymatic activity with the control group, but significant down-regulation in CYP1A2 enzymatic activity after being combined with paeoniae. The compound and its combined administration showed the inhibitory effect on CYP2B6 enzymatic activity, particularly being combined with angelicae. The compound showed identical effect with the four single components in terms of CYP1A2 mRNA expression and enzymatic activity.

Safety evaluations of the processed lateral root of Aconitum carmichaelii Debx. And its hepatotoxicity mechanisms in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: The processed lateral root of Aconitum carmichaelii Debx. is known as Fuzi, an extensively used Traditional Chinese Medicine to treat cardiovascular diseases, rheumatism arthritis, bronchitis, pains, and hypothyroidism, etc. Although Chinese Pharmacopeia regulates the safe clinical dosage of Fuzi at 3-15 g/person/day, such recommendation not only lacks bench evidence but also does not differentiate Fuzi with different processing types, such as Heishunpian and Paofupian. AIM OF THE STUDY: The current study aimed to 1) determine No-Observed-Adverse-Effect-Levels of Heishunpian and Paofupian in rats and 2) investigate the related toxicity mechanisms for their safe clinical use. MATERIALS AND METHODS: After giving clinically relevant dosing regimen of Heishunpian/Paofupian to rats, we conducted toxicity assessments including ECG monitoring, histopathological changes and serum biomarkers to detect organ injury. Metabolomic study in the liver revealed changes in endogenous metabolite levels after two-week treatment of Fuzi preparations or its corresponding six toxic alkaloids mixtures. RESULTS: The NOAEL for both bolus and two-week treatments of Heishunpian and Paofupian in rats was designated to be 7.5 g/kg and 15 g/kg, respectively. Corresponding recommended doses in humans were 7.5-25 g/person/day for Heishunpian and 15-50 g/person/day for Paofupian. Metabolic profiles revealed more significant alterations in endogenous substances from rats receiving the two Fuzi preparations than their corresponding toxic alkaloids mixtures. Upregulation of bile acid pathway could be responsible for Fuzi induced liver injury. CONCLUSIONS: Compared to the current maximum recommended dose, our suggested upper limit of guided dose for Heishunpian was comparable, whereas that for Paofupian could be further elevated. Both C19-diterpenoid alkaloids and co-occurring components in Fuzi preparations contributed to their hepatotoxicity via upregulation of bile acid pathway.

[Evaluation of P-glycoprotein mediated in vitro loperamide biliary excretion with sandwich-cultured rat hepatocytes model].

An in vitro P-glycoprotein mediated drug biliary excretion model (B-Clear model) was developed and validated using sandwich-cultured rat hepatocytes (SCRH) and a model substrate rhodamine 123 (Rh123). SCRH formed functional bile canalicular networks after 5 days of culture. Rh123 (10 micromol x L(-1)) was then incubated with the SCRH in standard Ca+ Hanks buffer or Ca(2+)-free buffer. The cumulative cell uptake and canalicular efflux of Rh123 under Ca2+ and Ca(2+)-free conditions were measured with a LC-MS/MS method. The biliary excretion index (BEI) and instinct biliary clearance (CL(bile, int)) were calculated. To assess the effect of known P-gp inhibitors on the efflux of Rh123, cyclosporine A (CyA), tariquidar (TQD) or quinidine (QND) (10, 50 and 100 micromol x L(-1)) was pre-incubated separately with SCRH for 30 min, then co-incubated with Rh123. The BEI and CL(bile, int) of Rh123 obtained from the SCRH model were (17.8 +/- 1.3) % and (10.7 +/- 0.9) mL x min(-1) x kg(-1), respectively. All the three P-gp inhibitors showed a dose-dependent inhibition on the bile clearance of Rh123, indicating that the B-Clear model with SCRH was functional properly. The biliary excretion of loperamide (LPAD) and the role of P-gp were further investigated with this validated model. The BEI and CL(bile, int) for LPAD (20 micromol x L(-1)) were obtained after it was incubated with SCRH for 30 min, and found to be (12.9 +/- 1.2)% and (6.1 +/- 0.3) mL x min(-1) x kg(-1) respectively. The dose-dependent inhibition on LPAD biliary excretion by CyA, TQD or QND confirmed the major role of P-gp in LPAD canalicular efflux. The results suggested that the B-Clear model with SCRH would be a useful tool for evaluation of P-gp mediated efflux and drug-drug interaction.

[Metabolic detoxification of bakuchiol is mediated by cytochrome P450 enzymes in human liver microsomes].

OBJECTIVE: To analyze cytochrome P450 (CYP) phenotyping for bakuchiol metabolism and study the mechanism of detoxification of bakuchiol by human liver microsomes (HLM) in vitro. METHODS: The CYP phenotyping for bakuchiol metabolism was determined using HLM combined with CYP specific inhibitors and recombinant human CYP isoforms. The relative activities of CYP isoforms were determined by analyzing the formation of the substrate metabolites using HPLC-MS/MS, in presence or absence of 1-aminobenzotriazole (ABT) which was CYP enzymes' broad spectrum inhibitor. The residual concentrations of bakuchiol in microsomal incubates were determined using HPLC to investigate ABT's effect on the metabolism of bakuchiol. The effects of CYP enzymes on the nephrotoxicity of bakuchiol were investigated using human kidney-2(HK-2) by MTT assay, in presence or absence of ABT. RESULTS: CYP1A2, CYP2C9, CYP2C19 and CYP3A4 in HLM were involved in bakuchiol metabolism, among which CYP2C19 showed the highest metabolic rate. Co-incubation with ABT (2.5 mmol/L) could inhibit more than 90% of the enzyme activities for CYP1A2, CYP2C9, CYP2C19 and CYP3A4. ABT (2.5 mmol/L) could inhibit the HLM metabolism of bakuchiol with inhibition ratio 83.24%±2.13%. When preincubated with ABT, the metabolic detoxification of bakuchiol by HLM was significantly reduced (P<0.05). CONCLUSION: The mechanism of metabolic detoxification of bakuchiol by HLM is associated with bakuchiol metabolism by CYP enzymes to form non toxic or lower toxic metabolites. The broad spectrum inhibitor of CYP could inverse the detoxification of HLM.

Metabonomic approaches investigate diosbulbin B-induced pulmonary toxicity and elucidate its underling mechanism in male mice.

Air Potato Yam is widely used in the treatment of many conditions such as cancer, inflammation, and goiter. Diosbulbin B (DIOB) is the primary active component of Air Potato Yam, and it exhibits anti-tumor and anti-inflammatory properties. The main purpose of this study was to determine the mechanism by which DIOB induces lung toxicity, using metabonomics and molecular biology techniques. The results showed that the lung toxicity induced by DIOB may occur because of a DIOB-induced increase in the plasma levels of long-chain free fatty acids and endogenous metabolites related to inflammation. In addition, treatment with DIOB increases the expression of the cyp3a13 enzyme, which leads to enhanced toxicity in a dose-dependent manner. The molecular mechanism underlying toxicity in mouse lung cells is the DIOB-mediated inhibition of fatty acid ß-oxidation, partial glycolysis, and the TCA cycle, but DIOB treatment can also compensate for the low Adenosine triphosphate (ATP) supply levels by improving the efficiency of the last step of the glycolysis reaction and by increasing the rate of anaerobic glycolysis. Using metabonomics and other methods, we identified the toxic effects of DIOB on the lung and clarified the underlying molecular mechanism.

Metabonomics reveals bisphenol A affects fatty acid and glucose metabolism through activation of LXR in the liver of male mice.

Bisphenol-A (BPA) is a representative environmental endocrine disrupting chemical that is widely used in the production of polycarbonate plastics and epoxy resins. Many studies have confirmed BPA to be closely associated with metabolic diseases, reproductive system diseases, and sex hormone-dependent cancers. In this study, we aimed to systematically elucidate the molecular action of BPA on liver fatty acid and glucose metabolism and the reasons for BPA-induced hypoglycemia through a metabonomics approach. C57BL/6 mice were orally treated with BPA (1, 10, 50, 250 µg/kg) for 35 days and the liver metabonomics and histopathology, molecular docking, mRNA expression levels and activities of enzymes were analyzed. Based on the high-resolution mass spectrometry (MS) for metabonomics and on various software and bioinformatic analysis methods, we found that BPA could affect fatty acid and glucose metabolism, block the TCA cycle, and BPA also regulated the nuclear receptor LXR caused hypoglycemia, thereby affecting the normal metabolic functions of the liver.

[Interaction between four herb compounds and a western drug by CYP3A4 enzyme metabolism in vitro].

OBJECTIVE: To explore the interaction between herbal medicines and western drugs based on CYP3A4 enzyme metabolism by using testotesrone as a probe in liver microsome metabolism system in vitro. METHOD: The mixed liver microsome enzymatic system consisting of rat liver microsomes by ultra-high-speed centrifuge was established. The substrate testosterone was added into the system and enzyme CYP3A4 metabolic activity was expressed by the output of 6beta-hydroxy-testosterone which was measured by HPLC method. The proper conditions for testotesrone metabolism in liver microsome system included substrate concentration, incubation time, pH and incubation temperature. When the conditions in vitro were determined, three kinds of Chinese herbal medicinal ingredients (Tetrahydropalmatine, neferine, panax notoginseng saponins) were diluted into different concentrations and incubated with testotesrone in the liver microsomes incubation system, respectively. The results were measured through metabolite production with or without the presence of Chinese medicines. We assessed the Chinese herbal medicinal ingredients effect on the metabolism of CYP3A4 enzyme through 6beta-hydroxy metabolite of testosterone production. RESULT: Liver microsomes were incubated in the system, the testosterone metabolited into 6beta-hydroxy testosterone. The metabolism conditions were proper at the concentration of testosterone 200 micromol x L(-1) which was incubated for 3.5 hours at 37 degrees C in pH 7.0, PBS 0.1 mol x L(-1). The inhibition of tetrahydropalmatine and panax notoginseng saponins on testotesrone were weak with IC50 > 100 micromol x L(-1). The neferine had a little inhibition on testotesrone metabolism, IC50 < 100 micromol L(-1). CONCLUSION: Tetrahydropalmatine and panax notoginseng saponins had no obvious effect on testotesrone metabolism. Neferine had a little effect on testotesrone metabolism. It prompted that drug-interaction could not be apparent between two kinds of Chinese medicines and the CYP3A4 enzyme substrate, Neferine could bring about drug-interaction.

Metabonomics Indicates Inhibition of Fatty Acid Synthesis, ß-Oxidation, and Tricarboxylic Acid Cycle in Triclocarban-Induced Cardiac Metabolic Alterations in Male Mice.

Triclocarban (TCC) has been identified as a new environmental pollutant that is potentially hazardous to human health; however, the effects of short-term TCC exposure on cardiac function are not known. The aim of this study was to use metabonomics and molecular biology techniques to systematically elucidate the molecular mechanisms of TCC-induced effects on cardiac function in mice. Our results show that TCC inhibited the uptake, synthesis, and oxidation of fatty acids, suppressed the tricarboxylic acid (TCA) cycle, and increased aerobic glycolysis levels in heart tissue after short-term TCC exposure. TCC also inhibited the nuclear peroxisome proliferator-activated receptor α (PPARα), confirming its inhibitory effects on fatty acid uptake and oxidation. Histopathology and other analyses further confirm that TCC altered mouse cardiac physiology and pathology, ultimately affecting normal cardiac metabolic function. We elucidate the molecular mechanisms of TCC-induced harmful effects on mouse cardiac metabolism and function from a new perspective, using metabonomics and bioinformatics analysis data.

Metabonomics reveals that triclocarban affects liver metabolism by affecting glucose metabolism, ß-oxidation of fatty acids, and the TCA cycle in male mice.

This study combined metabonomics with molecular biology techniques to identify differential endogenous substances produced by triclocarban (TCC) that affect plasma and liver metabolism in mice, to map their associated metabolic pathways, and to systematically determine the mechanism of TCC affecting liver metabolism in mice. The results showed that TCC affected liver metabolism by a mechanism involving the inhibition of glucose oxidation in the liver, promotion of anaerobic glycolysis and gluconeogenesis, and accelerated ß-oxidation of liver fatty acids and the TCA cycle, which lead to metabolic disorders of the liver microenvironment in mice. The analysis of endogenous substances in the liver and plasma indicated that TCC caused physiological and pathological changes in the liver, and affected the physiological state of mice and the metabolic balance of endogenous substances. Based on metabonomics and bioinformatics analysis methods, this study elucidated a new mechanism involved in how TCC affects liver metabolism.

[Studies on callus culture of Akebia trifoliata].

OBJECTIVE: To investigate the inductive factors of effect on Akebia trifoliata and establish culture method for A. trifoliata callus. METHOD: To study the possible effective factors of culture condition by comparing with different explantation, nutrient medium, pH, temperature, illumination, growth substance of plant and its ratio. RESULT: The inductivity of leaves was the highest about 87.5%, followed with the stem section and leafstalk; The inductivity of nutrient medium such as MS, B5 callus was higher than the ones like H, SH and the White callus amended one; It was found that low-grade Phvalue benefits the growth of callus. The experiment result showed that different pH showed little difference in quality. The best condition of culture was 25 degrees C in temperature. CONCLUSION: The best culture condition for callus was the leaves as explantation. The A. trifoliata callus culture's best inductive condition was MS +2, 4-D 4.0 mg x L(-1) + NAA 1.0 mg x L(-1) + KT 1.0 mg x L(-1) (pH 5.8), cultural temperature was 25 degrees C, cultivation was dark.

Tributyl phosphate impairs the urea cycle and alters liver pathology and metabolism in mice after short-term exposure based on a metabonomics study.

As a newly emerging environmental contaminant, tributyl phosphate (TBP) is of increasing concern because of the environmental problems it can cause. Studies have suggested that TBP induces hepatocellular adenomas and has malignant potential for hepatocellular carcinoma. However, the mechanisms of its adverse effects are unclear. In this study, metabonomic techniques were used to identify differential endogenous metabolites, draw network metabolic pathways and conduct network analysis to elucidate the underlying mechanisms involved in TBP induced pathological changes of the liver. The metabonomics study showed that TBP altered endogenous metabolites in the plasma and liver. The number of categories of endogenous metabolites with a VIP >1 were 14 in plasma and 20 in liver. The results also showed that TBP impaired urea synthesis in the liver. In addition, results of both in vitro and in vivo experiments indicated that TBP activated nuclear receptor CAR and inhibited CYP3a11 and CYP2b10 activities in the liver of mice after short-term exposure. These effects may be the underlying causes leading to TBP induced hepatocellular adenomas. This study combined metabonomics and other technical methods to clarify the mechanism of TBP-induced liver tumorigenesis from a new perspective.