Andrographolide ameliorates hepatic steatosis by suppressing FATP2-mediated fatty acid uptake in mice with nonalcoholic fatty liver disease.

Excessive intrahepatocellular lipid accumulation or steatosis is caused by abnormal lipid metabolism and a common character of nonalcoholic fatty liver disease (NAFLD), which may progress into cirrhosis and hepatocellular cancer. Andrographolide (Andro) is the primary active ingredient extracted from Andrographis paniculata, showing a protective role against dietary steatosis with the mechanism not fully understood. In this study, we showed that administration of Andro (50, 100, and 200 mg/kg/day for 8 weeks, respectively) attenuated obesity and metabolic syndrome in high-fat diet (HFD)-fed mice with improved glucose tolerance, insulin sensitivity, and reduced hyperinsulinemia, hyperglycemia, and hyperlipidemia. HFD-fed mice presented hepatic steatosis, which was significantly prevented by Andro. In vitro, Andro decreased the intracellular lipid droplets in oleic acid-treated LO2 cells. The selected RT-PCR array revealed a robust expression suppression of the fatty acid transport proteins (FATPs) by Andro treatment. Most importantly, we found that Andro consistently reduced the expression of FATP2 in both the oleic acid-treated LO2 cells and liver tissues of HFD-fed mice. Overexpression of FATP2 abolished the lipid-lowering effect of Andro in oleic acid-treated LO2 cells. Andro treatment also reduced the fatty acid uptake in oleic acid-treated LO2 cells, which was blunted by FATP2 overexpression. Collectively, our findings reveal a novel mechanism underlying the anti-steatosis effect of Andro by suppressing FATP2-mediated fatty acid uptake, suggesting the potential therapeutic application of Andro in the treatment of NAFLD.

Identification of GmGPATs and their effect on glycerolipid biosynthesis through seed-specific expression in soybean.

BACKGROUND: Genetic improvement of soybean oil content depends on in-depth study of the glycerolipid biosynthesis pathway. The first acylation reaction catalysed by glycerol-3-phosphate acyltransferase (GPAT) is the rate-limiting step of triacylglycerol biosynthesis. However, the genes encoding GPATs in soybean remain unknown. METHODS: We used a novel yeast genetic complementation system and seed-specific heterologous expression to identify GmGPAT activity and molecular function in glycerolipid biosynthesis. RESULTS: Sixteen GmGPAT genes were cloned by reverse transcription-PCR for screening in yeast genetic complementation. The results showed that GmGPAT9-2 could restore the conditional lethal double knockout mutant strain ZAFU1, and GmGPAT1-1 exhibited low acyltransferase activity in serial dilution assays. In addition, the spatiotemporal expression pattern of GmGPAT9-2 exhibited tissue specificity in leaves, flowers and seeds at different developmental stages. Furthermore, both the proportion of arachidic acid and erucic acid were significantly elevated in Arabidopsis thaliana transgenic lines containing the seed-specific GmGPAT9-2 compared wild type, but the oil content was not affected. CONCLUSION: Together, our results provide reference data for future engineering of triacylglycerol biosynthesis and fatty acid composition improvement through GPATs in soybean.

Transcriptome analysis reveals a composite molecular map linked to unique seed oil profile of Neocinnamomum caudatum (Nees) Merr.

BACKGROUND: Neocinnamomum caudatum (Nees) Merr., a biodiesel tree species in the subtropical areas of South China, India and Burma, is distinctive from other species in Lauraceae family and its seed oil is rich in linoleic acid (18:2) and stearic acid (18:0). However, there is little genetic information about this species so far. In this study, a transcriptomic analysis on developing seeds of N. caudatum was conducted in an attempt to discern the molecular mechanisms involving the control of the fatty acid (FA) and triacylglycerol (TAG) biosynthesis. RESULTS: Transcriptome analysis revealed 239,703 unigenes with an average length of 436 bp and 137 putative biomarkers that are related to FA formation and TAG biosynthesis. The expression patterns of genes encoding ß-ketoacyl-acyl carrier protein synthase I (KASI), ß- ketoacyl-acyl carrier protein synthase II (KASII), stearoyl-ACP desaturase (SAD), fatty acid desaturase 2 (FAD2), fatty acid desaturase 8 (FAD8) and acyl-ACP thioesterase A/B (FATA/B) were further validated by qRT-PCR. These genes displayed a very similar expression pattern in two distinct assays. Moreover, sequence analysis of different FATBs from diverse plant species revealed that NcFATB is structurally different from its counterpart in other species in producing medium-chain saturated FAs. Concertedly, heterologous expression of NcFATB in E. coli BL21 (DE3) strain showed that this corresponding expressed protein, NcFATB, prefers long-chain saturated fatty acyl-ACP over medium-chain fatty acyl-ACP as substrate. CONCLUSIONS: Transcriptome analysis of developing N. caudatum seeds revealed a composite molecular map linked to the FA formation and oil biosynthesis in this biodiesel tree species. The substrate preference of NcFATB for long-chain saturated FAs is likely to contribute to its unique seed oil profile rich in stearic acid. Our findings demonstrate that in the tree species of Lauraceae family, the FATB enzymes producing long-chain FAs are structurally distinct from those producing medium-chain FAs, thereby suggesting that the FATB genes may serve as a biomarker for the classification of tree species of Lauraceae family.

[Application of GFH in the municipal effluent reuse for landscape].

In order to prevent increased algae growth for the reuse of reclaimed water as landscape water, the removal mechanism of phosphate, DOM and nitrogen by adsorption onto GFH was studied. The result showed that the removal rate of phosphate was the highest, when the influent concentration of TP was 0.059-0.725 mg/L, and the concentration of PO4(3-) -P was 0.004-0.684 mg/L, the effluent concentration of TP was less than 0.05 mg/L(removal rate was 91.1%), and of PO4(3-) -P was less than 0.023 mg/L(removal rate was 95.4%) by adsorption onto GFH. GFH could remove macromolecular DOM (mainly fulvic acids) preferentially, and removal rate of DOM was 28.5%, but which would improve the aromaticity proportion of DOM. Because of the strong oxidizing property of GFH and O3, NH4(+) -N and NO2(-) -N occurred nitrification in reclaimed water, the average removal rates of NH4(+) -N and NO2(-) -N were 37.3% and 59% respectively.

[Autotrophic nitrogen removal and enhanced biological phosphorus removal from municipal wastewater in a three-sludge system].

Using a three-sludge system consisted of anaerobic/oxic (A/O) process, partial nitritation and anaerobic ammonium oxidation (ANAMMOX) reactors, cost-effective removal of nitrogen and phosphate from municipal wastewater was achieved. The experimental results showed that effluent total phosphorus (TP) of the A/O system was less than 0.5 mg/L under hydraulic retention time (HRT) of 3.6 h. Partial nitritation with nitrite accumulation efficiency of 75% -96% was realized in the partial nitritation system under room temperature, DO < 0.2 mg/L and HRT of 4.6 h. Under temperature of 27-30 degrees C and HRT of 1.4 h, effluent total nitrogen (TN) and TN removal rate of ANAMMOX reactor were less than 8 mg/L with the minimum value of 1.6 mg/L and 0.57 kg/(m3 x d), respectively. In the three-sludge system, phosphate accumulating organisms, ammonia-oxidizing bacteria and Anammox bacteria existed under suitably environmental condition to optimize the microbial community structure and improve treatment efficiency of various units. Autotrophic nitrogen removal can reduce 62.5% of the oxygen supply, save 100% of denitrification carbon sources theoretically, lower the sludge production, and greatly decrease carbon dioxide emission. As compared to traditional biological nutrient removal process, the three-sludge system has great advantages and potential in energy saving and carbon dioxide emission reduction to realize sustainable development of water resources.