In-vitro Metabolism of a Novel Monocrotophos Derivative by Rat and Japanese Quail

NADPH-dependent inhibition of hepatic microsomal carboxylesterase by a derivative of monocrotophos (coded as RPR-5) was studied in rat and Japanese quail as a measure of monooxygenase-catalysed activation of RPR-5. There was NADPH-dependent inhibition of hepatic microsomal α-naphthyl acetate esterase (carboxylesterase) both in rat and quail, indicating monooxygenase-catalysed formation of an oxon that subsequently phosphorylated α-NaE. The pattern of in-vitro metabolism of ¹⁴C-labelled RPR-5 by 11000g supernatant (11-S), microsomes and 105000g supernatant (105-S) fractions of rat and quail livers suggested the involvement of microsomal monooxygenases and carboxylesterases. A radiolabelled metabolite (M2) was tentatively identified as an acid produced by carboxyl esterase attack. In rat, metabolism by microsomal and cytosolic (105-S) carboxylesterases appeared to predominate with relatively little oxidative metabolism. In quail, putative microsomal carboxylesterase hydrolysis of RPR-5 was much lower than in the rat with almost neglible hydrolysis by cytosolic fractions. Also, production of M₂ by quail microsomes was substantially reduced after addition of NADPH, suggesting inhibition of a carboxyl esterase by the oxon of RPR-5. Differences in this detoxification of RPR-5 between rat and quail may be an important factor in determining selective toxicity and the results underline the importance of relating metabolism to toxicity when selecting animal models for toxicity testing.     

脂肪细胞培养研究现状

脂肪细胞培养为研究脂肪代谢紊乱疾病提供了一种有效的途径。基质血管成分系统的研究形成了完整的前脂肪细胞理论,随后脂肪细胞原代培养技术得到了广泛的研究,国外各种家畜的脂肪细胞培养模型得以建立,而国内在脂肪细胞培养方面的研究起步较晚,且多集中于人及鼠的脂肪细胞培养。脂肪细胞的传代培养在单一因子对脂肪细胞的功能研究上有一定的优越性,尤其对不宜于在活体采样的人及动物。脂肪细胞培养的另一个研究方向是脂肪细胞与其他组织细胞的联合培养,对研究机体内各种组织细胞间的相互作用提供了良好的基础。     

发酵乳对肥胖小鼠肝脂代谢和肠道菌群的影响

为分析发酵乳对肥胖小鼠肝脂代谢和肠道菌群的影响,本研究将60只C57BL/6小鼠随机分为5组：正常对照组（CON）、模型组（MD）、易善复组（YG）、低浓度发酵乳干预组（LFG）和高浓度发酵乳干预组（HFG）。肥胖模型造模成功后进行为期6周的灌胃,试验结束测量小鼠的空腹体重和身长,计算LEE’S指数,采血检测生化指标,采集肝脏组织观察病理变化;采集小鼠盲肠内容物进行肠道菌群测序。结果表明：1）与CON相比,MD小鼠体重、LEE’S指数、ALT和AST均极显著提升（P<0.01）,表明肥胖模型建立成功。2）与MD相比,发酵乳干预组小鼠体重、LEE’S指数、ALT和TC均极显著下降（P<0.01）,其中AST和TG显著下降（P<0.05）,同时发酵乳和易善复组小鼠肝组织脂肪变性有明显好转,基本没有明显的脂肪空泡。表明发酵乳可以抑制肥胖小鼠体重的增加,改善肥胖小鼠的肝脂代谢功能。3）与MD相比干预组小鼠肠道菌群的多样性水平有一定程度的恢复,尤其是厚壁菌门和拟杆菌门。表明发酵乳对于小鼠肠道菌群多样性的恢复有一定的作用。综上,本研究阐明了发酵乳对肥胖小鼠肝脂代谢和肠道菌群的影响,为功能型发酵乳的开发提供理论依据。     

Pharmacokinetics and Metabolism Study of Deep-Sea-Derived Butyrolactone I in Rats by UHPLC–MS/MS and UHPLC–Q-TOF-MS

Butyrolactone I (BTL-I) is a butanolide isolated from the deep-sea-derived fungus, Aspergillus sp. It provides a potential new target for the prevention and treatment of food allergies. This study aimed to investigate the metabolic and pharmacokinetic profile of BTL-I in rats. The metabolic profiles were obtained by UHPLC–Q-TOF-MS. As a result, eleven metabolites were structurally identified, and the proposed metabolic pathways of BTL-I were characterized. The main metabolites were the oxidative and glucuronidative metabolites. In addition, a sensitive UHPLC–MS/MS method was established for the quantitation of BTL-I in rat plasma (LOQ = 2 ng/mL). The method was fully validated and successfully applied to the pharmacokinetic study of BTL-I in rats after oral administration or intravenous administration. The oral bioavailability was calculated as 6.29%, and the maximum plasma concentrations were 9.85 ± 1.54 ng/mL and 17.97 ± 1.36 ng/mL for intravenous and intragastric dosing groups, respectively.     

不同浓度玉米生长调节剂对玉米氮代谢的影响

采用田间小区试验,研究了不同浓度新型玉米生长调节剂对玉米氮代谢的影响．结果表明,不同浓度玉米生长调节剂对灌浆期玉米功能叶、茎秆中氮素的转运,功能叶硝酸还原酶与蛋白水解酶活性,功能叶和籽粒的谷氨酰胺合成酶与转化酶活性均有明显的促进作用,显著增加玉米产量、籽粒蛋白质含量及籽粒蛋白质产量．在调节剂的3个浓度中,以PGRCl较合理．PGRCl处理玉米籽粒的蛋白氮含量比对照提高0．83g／kg,籽粒产量比对照增加13．18％,籽粒蛋白质产量比对照增加19．59％．     

转Pi9抗稻瘟病基因水稻株系的比较转录组分析

【目的】在转录水平上解析Pi9基因介导的稻瘟病抗性调控机理,为培育抗病水稻品种提供理论依据。【方法】向水稻品种日本晴(NPB)及其转Pi9抗稻瘟病基因株系(NPB/Pi9)接种稻瘟菌。分别于接种后0 h、12 h、24 h、36 h提取叶组织样品,选取12503个水稻基因定制基因芯片,进行水稻基因转录组分析,并通过qRT-PCR对部分差异表达基因进行验证。【结果】NPB/Pi9在接种后12 h、24 h和36 h的基因表达量分别与其接种0 h表达量比较,共检测到7754个差异表达基因;相应地,感病水稻NPB在以上时间点共检测到7385个差异表达基因;在接种后36 h,NPB/Pi9的差异表达基因数目显著多于NPB。比较NPB/Pi9和NPB相同时间点的基因表达量,共获得4065个差异表达基因,其中接种后36 h的差异表达基因显著多于接种后0 h、12 h或24 h。因此,NPB/Pi9的稻瘟病防御反应更强烈。对NPB/Pi9与NPB相同时间点的差异表达基因进行GO和KEGG分析,细胞外区域、植物对刺激应答、转录调控、氧化还原、离子结合、次生代谢和植物激素相关的GO分类在接种后呈显著富集,苯丙氨酸代谢、类黄酮生物合成和植物激素信号途径的KEGG通路在接种后显著富集。与效应分子触发的免疫反应(ETI)相关的水杨酸信号途径、几丁质酶,以及与病原相关分子模式触发的免疫反应(PTI)相关的胞外区域、对刺激的应答、木质素合成等,均在抗感水稻之间差异表达。而且PTI/ETI共有的WRKY转录因子、MAPK激酶、茉莉酸和乙烯信号途径等发生差异表达。综上所述,NPB/Pi9和NPB的差异表达模式与ETI和PTI相关,两者相互联系并在Pi9介导的稻瘟病抗性中发挥作用。【结论】与日本晴比较,抗病基因型NPB/Pi9对稻瘟病防御反应更强烈。转录因子、激酶、NBS-LRR基因、几丁质酶、水杨酸、茉莉酸和乙烯信号途径,以及植物次生代谢在Pi9介导的稻瘟病抗病反应中发挥重要作用。     

Evaluation of Intestinal Absorption of Dietary Halocynthiaxanthin,a Carotenoid from the Sea Squirt Halocynthia roretzi

Halocynthiaxanthin is an acetylenic carotenoid mainly found in Halocynthia roretzi. To date, several bioactivities of halocynthiaxanthin have been reported, but its mechanism of digestion and absorption in mammals has not been studied yet. In this study, we evaluated the intestinal absorption of halocynthiaxanthin in mice. The halocynthiaxanthin-rich fraction was prepared from the tunicate Halocynthia roretzi. Mice were orally administered the fraction at a dose of 5 mg/kg body weight. The halocynthiaxanthin levels in the plasma, liver, and small intestine, were quantified using HPLC-PDA, 1, 3, 6, and 9 h after ingestion. The halocynthiaxanthin-rich fraction mainly consisted of the all-trans form and a small amount of cis forms. These three isomers were detected in the plasma of mice 3 h after ingestion. Time-course changes after the ingestion of this fraction were found, with cis isomers being more abundant than the all-trans isomer in the mouse plasma and liver. In the small intestine, however, the all-trans isomer was primarily detected. The possibility that cis isomers might be absorbed rapidly from the small intestine cannot be denied, but our results suggest that dietary all-trans-halocynthiaxanthin might be isomerized to the cis isomer after intestinal absorption.     

茶足柄瘤蚜茧蜂脂代谢相关的滞育关联基因差异表达分析

本文对茶足柄瘤蚜茧蜂滞育组与正常发育组进行转录组测序,筛选差异表达基因,根据功能注释发现这些滞育关联基因主要与碳水化合物代谢、脂质代谢以及信号转导等途径相关。借助KEGG数据库,共筛选出滞育期间401个与脂代谢相关的差异基因,在滞育期间呈现不同程度的上调表达,涉及了脂肪酸生物合成、甘油脂代谢、类固醇生物合成等代谢途径,除与脂质合成相关的脂肪酸合成酶、超长链脂肪酸延伸酶基因上调表达外,与脂质分解相关的酶,如酯酶同样上调表达,可能与提高茶足柄瘤蚜茧蜂免疫力有关;与激素合成相关基因的上调表达,可能抑制茶足柄瘤蚜茧蜂卵巢发育。     

Effects of different active constituents of Gynostemma pentaphyllum on mitochondrial energy metabolism-related proteins in endothelial cells induced by ox-LDL

AIM To investigate the effects of different components of Gynostemma pentaphyllum ［gypenosides (Gps), gypenoside XLIX (GpXLIX) and ginsenoside Rb3 (GRb3)］ on mitochondrial energy metabolism-related proteins in endothelial cells induced by oxidized low-density lipoprotein (ox-LDL). METHODS EA.hy926 cells were divided into control group, model group, Gps group, GpXLIX group and GRb3 group. The cells in control group were cultured only in DMEM complete medium. The cells in model group were treated with 100 mg/L ox-LDL for 48 h. The cells in Gps group, GpXLIX group and GRb3 group were treated with 100 mg/L ox-LDL for 24 h, and then treated with Gps, GpXLIX and GRb3 at 100 mg/L for another 24 h, respectively. The ATP content in each group was detected by ELISA. The expression levels of mitochondrial energy metabolism-related proteins, cytochrome C oxidase subunit 5a （Cox5a）, NADH:ubiquinone oxidoreductase core subunit S1 （Ndufs1）, ATP synthase F1 subunit alpha (ATP5a) and cytochrome C (Cyt C）, were determined by Wes automatic Western blot quantitative analysis system and Western blot. RESULTS Compared with control group, the ATP content in model group was decreased (P<0.01). After drug intervention, the ATP content increased to different degrees in Gps group, GpXLIX group and GRb3 group (P<0.01). The results of Wes automatic Western blot quantitative analysis system were consistent with those of Western blot. These results showed that compared with control group, the protein expression of Cox5a, Ndufs1 and ATP5a in model group was decreased, and the protein expression of Cyt C was increased (P<0.01). After intervention, the protein expression of Cox5a, Ndufs1 and ATP5a was increased and the protein expression of Cyt C was decreased in Gps group, GpXLIX group and GRb3 group (P<0.05 or P<0.01). Among them, the effect of Gps on the protein expression of Cox5a, Ndufs1 and Cyt C was significantly stronger than those of the 2 monomer components, and the effect of GRb3 was found to be superior in the 2 monomer components. The effect of GpXLIX on ATP5a protein was superior to the other 2 components. CONCLUSION Gynostemma total saponins and related active ingredients protect ox-LDL-induced endothelial cells by affecting mitochondrial energy metabolism-related proteins, thereby preventing and treating atherosclerosis.