摘要：
Previous studies show that several pathways are involved in sanguinarine-induced apoptotic cell death, including Ala downregulation, inhibition of NF-kB activation, mediation of ROS production, downregulation of anti-apoptosis proteins XIAP and clAP-1, upregulation of BAX, and downregulation of BCL2. In this study, we found out that the quenching of ROS generation by N-acetyl-L-cysteine (NAC), a scavenger of ROS, reversed sanguinarine-induced apoptosis effects, also we found out that sanguinarine-induced rat hepatic stellate T6 cells (HSC-T6 cells) apoptosis was correlated with the generation of increased ROS, which was followed by the activation of caspase-8 (-3, -6, and -9), and the decreasing in the miltochondrial membrane potential (MMP) and the down-regulation of anti-apoptotic protein Bcl-2. It is not clear whether BCL2's downregulation relates to its promoter methylation and miR-15a/16-1 expression which can bind to BCL2 3'-UTR (un-translation reagon). We showed that sanguinarine-induced down regulation of BCL2 was associated with the increased methylation rate of BCL2 promotor district and the increased expression of miR-15a/16-1. HSC-T6 cells treatment with 5-Aza-2'-deoxycytidine (5'-Aza-CdR) impeded sanguinarine-induced BCl2 promotor district methylation and recovered BCL2's expression. Over expression of BCL2 using pEGFP-N1 vector decreased sanguinarine-induced HSC-T6 cells apoptotic death significantly but not completely. These observations clearly showed that BCl2 down regulation was associated with its promoter methylation and miR-15a/16-1 upregulation in sanguinarine-induced Rat HSC-T6 cells. (C) 2014 Japanese Pharmacological Society. Production and hosting by Elsevier B.V.
摘要：
Background: Sanguinarine (SA) is a quaternary benzo[c]phenanthridine alkaloid that is mainly present in the Papaveraceae family. SA has been extensively studied because of its antimicrobial, anti-inflammatory, antitumor, antihypertensive, antiproliferative and antiplatelet activities. Metabolic studies demonstrated that SA bioavailability is apparently low, and the main pathway of SA metabolism is iminium bond reduction resulting in dihydrosanguinarine (DHSA) formation. Nevertheless, the metabolic enzymes involved in SA reduction are still not known in detail. Thus, the aim of this study was to investigate the rat liver microsomes and cytosol-induced SA iminium bond reduction, and to examine the effects of cytosol reductase inhibitors on the reductive activity. Methods: DHSA formation was quantified by HPLC. The possible enzymes responsible for DHSA formation were examined using selective individual metabolic enzyme inhibitors. Results: When SA was incubated with liver microsomes and cytosol in the absence of NAD(P)H, DHSA, the iminium bond reductive metabolite was formed. The reductase activity of the liver microsomes and cytosol was also enhanced significantly in the presence of NADH. The amount of DHSA formed in the liver cytosol was 4.6-fold higher than in the liver microsomes in the presence of NADH. The reductase activity in the liver cytosol was inhibited by the addition of flavin mononucleotide and/or riboflavin. Inhibition studies indicated that menadione, dicoumarol, quercetin and 7-hydroxycoumarin inhibited rat liver cytosol-mediated DHSA formation in the absence of NADH. However, only menadione and quercetin inhibited rat liver cytosol-mediated DHSA formation in the presence of NADH. Conclusions: These results suggest that the SA iminium bond reduction proceeds via two routes in the liver cytosol. One route is direct non-enzymatic reduction by NAD(P)H, and the other is enzymatic reduction by possible carbonyl and/or quinone reductases in the liver cytosol.
摘要：
A Gram-negative, rod-shaped bacterial strain named PB-CS01 which is bioluminescence-positive was isolated from contaminated commercial pork that probably had been exposed to seafood. Optimal growth of strain PB-CS01 requires the presence of 3.0% (w/v) NaCl and a temperature of 20 degrees C. Phylogenetic analysis based on 16S rRNA gene sequences of strain PB-CS01 and other Photobacterium species showed that the novel isolate belongs to the genus Photobacterium. Sequence similarity analysis between PB-CS01 and other species indicates that the closest relatives of strain PB-CS01 are Photobacterium phosphoreum ATCC 11040 (99.9%), Photobacterium kishitanii pjapo.1.1 (99.8%) and Photobacterium iliopiscarium ATCC 51760 (99.5%). The most abundant fatty acids were summed feature 3 (50.77%; C-16:1 omega 7c and/or C-16:1 omega 6c) and C-16:0 (15%). The fatty acid profile is similar to that of the genus Photobacterium but this report is the first to describe C-16:1 omega 6c as one of the compositions of summed feature 3 of the genus Photobacterium. The G+C content of the genomic DNA of strain. PB-CS01 was 44.8 mol%. Overall, strain. PB-CS01 is a novel Photobacterium species.
作者机构：
[Bai Xia; Xiao Hong-bo; Mai Pei; Sun Zhi-liang; Wang Shui-lian; Liu Zhao-ying; Dong Wei; Chen Xiao-jun; Wang Hui; Cai Jie] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[Bai Xia; Xiao Hong-bo; Mai Pei; Sun Zhi-liang; Liu Zhao-ying; Dong Wei; Chen Xiao-jun; Wang Hui; Cai Jie] Engn Res Ctr Vet Drugs Hunan Prov, Changsha 410311, Hunan, Peoples R China.
通讯机构：
[Sun Zhi-liang] H;Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.
关键词：
angiopoietin-like protein 3;clone;pregnancy toxemia;goat
摘要：
Pregnancy toxemia is a metabolic disorder of lipid and glucose. Recent investigations have found that angiopoietin-like protein 3 (ANGPTL3) can contribute to disorder of carbohydrate and lipid metabolism. The present study was conducted to investigate the change of ANGPTL3 expression during pregnancy toxemia. We firstly cloned the full-length cDNA of ANGPTL3 in Liuyang Black goats, revealing that goat ANGPTL3 had the typical structure of the angiopoietin-like family, and its mRNA was exclusively expressed in liver. Pregnancy toxemia of pregnant goat does with twins during late gestation was induced by being fasted for 72 h, and then they were recovered after 5 d of refeeding. Hepatic ANGPTL3 gene expression was significantly down-regulated concomitantly with decreased serum glucose concentration, elevated serum β-hydroxybutyrate and free fatty acid levels in pregnant toxemic goats, and these changes were reversed after refeeding. These results suggest ANGPTL3 may play a certain role in the development of pregnancy toxemia in goats.
作者：
Zhang, D. S.;Liu, Z. Y.;Chen, X. J.;Kuang, G. W.;Pan, R. T.;...
期刊：
AFRICAN HEALTH SCIENCES,2012年12(4):522-529 ISSN：1680-6905
通讯作者：
Sun, Z. L.
作者机构：
[Chen, X. J.; Sun, Z. L.; Zhang, D. S.; Pan, R. T.; Liu, Z. Y.] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.;[Kuang, G. W.] Inst Hunan Prov Vet Drugs & Feed Supervis, Changsha 410006, Hunan, Peoples R China.;[Sun, Z. L.] Hunan Agr Univ, Changsha 410128, Hunan, Peoples R China.
通讯机构：
[Sun, Z. L.] H;Hunan Agr Univ, Changsha 410128, Hunan, Peoples R China.
关键词：
Angiopoietin-like protein 3 (ANGPTL3);DNA methylation;Pigs
摘要：
Background: Angiopoietin-like protein 3 (ANGPTL3), a member of angiopoietin-like proteins, has been demonstrated to affect lipid metabolism by inhibiting the activity of lipoprotein lipase (LPL). Objective: To compare the ANGPTL3 mRNA and protein expression, exon mutation and promoter district CpG island methylation state between Ningxiang Pigs and Changbai Pigs. Methods: Pigs were slaughtered and about 100 mg of tissue samples and subcutaneous adipose tissue were collected and stored for analysis. Quantitative Real-Time PCR, Western blotting, exons sequencing, and HRM analysis were carried out. Results: ANGPTL3 was expressed in liver but not in fat and lean meat. Compared with Changbai pigs, ANGPTL3 mRNA level in Ningxiang pigs was lower. However, the protein expression showed no difference between these two groups of pigs. Sequences analysis showed that four variations existed between Changbai Pigs and Ningxiang Pigs, among which three variations caused no change of amino acids, and the other one caused amino acid mutation from Val (Changbai) to Met (Ningxiang). The ANGPTL3 promoter district CpG island bisulfite-PCR and sequencing results showed that the mean methylation rate ranged from 70.952% to 95.238% between Changbai pig and Ningxiang pig (p<0.05). Conclusion: These results support the significant difference (p<0.05) between Changbai pig and Ningxiang pig in high-resolution melting (HRM) analysis. All these results may be helpful for a better understanding of the role of ANGPTL3 in lipid metabolism.
摘要：
In this study, the stress degradation of olaquindox under conditions of hydrolysis (neutral, acidic and basic), oxidation and photolytic stress was investigated. In order to characterize each degradation product, we developed a rapid, sensitive and reliable high-performance liquid chromatography combined with hybrid ion trap/time-of-flight mass spectrometry (LC/MS-IT-TOF) method. The degradation products formed under different forced conditions were separated using an ODS-C18 column with gradient elution. Multiple scans of degradation products in MS and MS/MS modes and accurate mass measurements were performed through data-dependent acquisition. The structural elucidations of degradation products were performed by comparing the changes in the accurate molecular masses and fragment ions generated from precursor ions with those of parent drug. The present results showed that maximum degradation was observed in hydrolysis, especially in the acidic condition. The drug was also degraded significantly under photolytic conditions. A total of 12 degradation products of olaquindox were detected and characterized using the developed method. The main degradation product was formed by the complete cleavage of side chain to form 3-methyl-2-hydroxylquinoxaline-4-oxide. A degradation pathway of olaquindox was also tentatively proposed for the first time based on these characterized structures. (C) 2011 Elsevier B.V. All rights reserved.
