Antrodia cinnamomea and its compound dehydroeburicoic acid attenuate nonalcoholic fatty liver disease by upregulating ALDH2 activity.

ETHNOPHARMACOLOGICAL RELEVANCE: Nonalcoholic fatty liver disease (NAFLD) is a prevalent liver disease, but currently has no specific medication in clinic. Antrodia cinnamomea (AC) is a medicinal fungus and it has been shown that AC can inhibit high fat diet (HFD)-induced lipid deposition in mouse livers, but the effective monomer in AC and mechanism against NAFLD remain unclear. It has been reported that aldehyde dehydrogenase 2 (ALDH2) activation shows protective effects on NAFLD. Our previous study demonstrates that AC and its monomer dehydroeburicoic acid (DEA) can upregulate the ALDH2 activity on alcoholic fatty liver disease mouse model, but it is not clear whether the anti-NAFLD effects of AC and DEA are mediated by ALDH2. AIM TO STUDY: To elucidate the active compound in AC against NAFLD, study whether ALDH2 mediates the anti-NAFLD effects of AC and its effective monomer. MATERIALS AND METHODS: WT mice, ALDH2-/- mice and ALDH2-/- mice re-expressed ALDH2 by lentivirus were fed with a methionine-choline deficient (MCD) diet or high fat diet (HFD) to induce NAFLD, and AC at the different doses (200 and/or 500 mg/kg body weight per day) was administrated by gavage at the same time. Primary hepatocytes derived from WT and ALDH2-/-mice were stimulated by oleic acid (OA) to induce lipid deposition, and the cells were treated with AC or DEA in the meantime. Lentivirus-mediated ALDH2-KD or ALDH2-OE were used to knock down or overexpress ALDH2 expression in HepG2 cells, respectively. Finally, the effects of DEA against NAFLD as well as its effects on upregulating liver ALDH2 and removing the harmful aldehyde 4-hydroxynonenal (4-HNE) were studied in the MCD diet-induced NAFLD mouse model. RESULTS: In WT mice fed with a MCD diet or HFD, AC administration reduced hepatic lipid accumulation, upregulated ALDH2 activity in mouse livers, decreased 4-HNE contents both in mouse livers and serum, inhibited lipogenesis, inflammation and oxidative stress and promoted fatty acid ß-oxidation. These effects were abolished in ALDH2 KO mice but could be restored by re-expression of ALDH2 by lentivirus. In primary hepatocytes of WT mice, AC and DEA inhibited OA-induced lipid accumulation and triglyceride (TG) synthesis, promoting the ß-oxidation of fatty acid in the meantime. However, these effects were lost in primary hepatocytes of ALDH2 KO mice. Moreover, the expression level of ALDH2 significantly affected the inhibitory effects of AC and DEA on OA-induced lipid deposition in HepG2 cells. The effects of AC and DEA on suppressing lipid deposition, inhibiting mitochondrial ROS levels, reducing TG synthesis, and promoting ß-oxidation of fatty acid were all enhanced with the overexpression of ALDH2 and reduced with the knockdown of ALDH2 expression. DEA showed dose-dependent effects on inhibiting liver lipid deposition, elevating ALDH2 activity and reducing 4-HNE levels in the livers of MCD diet-induced NAFLD mice. CONCLUSION: DEA is the effective compound in AC against NAFLD. The related anti-NAFLD mechanisms of AC and DEA were through upregulating ALDH2 expression and activity, thus enhancing the elimination of 4-HNE in the livers, and sequentially alleviating oxidative stress and inflammation, promoting fatty acid ß-oxidation and decreasing lipogenesis.

Protective Effects of Antrodia cinnamomea and Its Constituent Compound Dehydroeburicoic Acid 32 Against Alcoholic Fatty Liver Disease.

BACKGROUND: Alcoholic fatty liver disease (AFLD), a leading chronic hepatic disease, affects an increasing number of people, and no effective drugs for the treatment of AFLD are available. Antrodia cinnamomea (AC) can inhibit AFLD, but its mechanisms and the effective compound in AC are unknown. OBJECTIVE: We aimed to explore the anti-AFLD mechanism of AC and the active compound within AC. METHODS: Wild-type (WT) C57BL/6J mice underwent 4 weeks of daily ethanol (EtOH) feeding to induce AFLD. AC or dehydroeburicoic acid 32 (DEA32), a compound in AC, was given to the mice. Parallel experiments to assess the effect of AC were conducted in aldehyde dehydrogenase 2 (ALDH2)-knockout (KO) mice. Primary mouse hepatocytes were incubated with ethanol and Alda- 1 (an ALDH2 agonist), AC or DEA32. RESULTS: In WT mice with AFLD, AC reduced lipid deposition, increased the expression and activity of ALDH2, reduced the acetaldehyde content, and downregulated the expression of lipogenic and inflammatory genes in the liver. These effects of AC disappeared in ALDH2 KO mice. DEA32 was identified as an active compound in AC, as its effects on EtOH-treated WT hepatocytes were similar to those of AC, which were comparable to the effects of Alda-1. These effects of DEA32 disappeared in EtOH-treated ALDH2 KO hepatocytes. Furthermore, in WT mice with AFLD, DEA32 reduced lipid deposition, increased the activity of ALDH2, and reduced the accumulation of acetaldehyde in the liver. DEA32 also downregulated the mRNA expression of genes related to lipogenesis and inflammation. CONCLUSION: AC and its constituent compound DEA32 inhibit AFLD by upregulating ALDH2 activity, accelerating acetaldehyde metabolism, and suppressing lipogenesis and inflammation in the liver.

Polyurethane Nanoparticle-Loaded Fenofibrate Exerts Inhibitory Effects on Nonalcoholic Fatty Liver Disease in Mice.

Polyurethane (PU) nanoparticles are potential drug carriers. We aimed to study the in vitro and in vivo efficacy of biodegradable PU nanoparticles loaded with fenofibrate (FNB-PU) on nonalcoholic fatty liver disease (NAFLD). FNB-PU was prepared by a green process, and its preventive effects on NAFLD were investigated on HepG2 cells and mice. FNB-PU showed sustained in vitro FNB release profile. Compared to FNB crude drug, FNB-PU significantly decreased triglyceride content in HepG2 cells incubated with oleic acid and in livers of mice with NAFLD induced by a methionine choline deficient diet, and increased plasma FNB concentration of the mice. FNB-PU increased absorption of FNB and therefore enhanced the inhibitory effects of FNB on NAFLD.

[Characteristics of nitrogen transportation in Castanea mollissima forest around the Fushi Reservoir watershed in north Zhejiang Province, China.] / æµ™æ±Ÿèµ‹çŸ³æ°´åº“é›†æ°´åŒºæ¿æ —æž—åœŸå£¤æ°®ç´ è¿ç§»ç‰¹å¾.

In this study, through setting runoff plots in Chinese chestnut (Castanea mollissima) fo-rest, the runoff water samples were collected and tested from January to October, 2013. Combining with soil nitrogen data before and after the rainy season, we analyzed the characteristics of soil nitrogen transportation in chestnut forest around the Fushi Reservoir watershed in north Zhejiang Pro-vince, China. The results showed that among the 10 surface runoff events of Chinese chestnut fo-rest, there exhibited a significant difference in runoff water volume of each runoff plot, and the maximum amount was 0.51 m3. Nitrogen was the major element associated with nutrient loss in the study region. The concentrations of nitrate nitrogen (NO3--N), ammonium nitrogen (NH4+-N), total nitrogen (TN) andchemical oxygen demend (CODMn) in the runoff water ranged from 0.02-1.87, 0.04-3.53, 1.69-5.33 and 5.30-14.07 mg·L-1, respectively. The water quality indexes were greatly affected by the runoff volume, and the relationship between the amount of nitrogen loss and runoff could be well fitted by using a linear equation. The difference in nitrogen contents of the soil in the upper, middle and lower part of the runoff plots was evident both before and after the rainy season. Moreover, the nitrogen content increased with the increasing altitude, but this trend decreased with increasing soil depth. Comparison of the difference between the four forms of soil nitrogen (NO3--N, NH4+-N, TN and hydrolyzable nitrogen Hydro-N of soil) before and after the rainy season showed that there existed significant differences in Hydro-N and TN, and the average diffe-rence values were 20.21 and 307.49 mg·kg-1, respectively. In this area, there was a great risk of nitrogen loss with runoff, and the potential non-point source pollution in Fushi Reservoir should be concerned.

Recombinant high-density lipoproteins and their use in cardiovascular diseases.

The unique anti-atherosclerosis abilities and other cardioprotective properties make high-density lipoprotein (HDL) a promising solution in treating cardiovascular diseases. A number of studies showed that HDL-based therapy was well tolerated and has great potential in the future. Among all these new agents, the most studied ones including recombinant HDL, recombinant human apolipoproteins, apolipoprotein mimetic peptides and recombinant HDL used as contrast agents in cardiovascular imaging are discussed here. Recombinant HDL and apolipoproteins are promising in diagnosing and treating cardiovascular diseases.

G5-PEG PAMAM dendrimer incorporating nanostructured lipid carriers enhance oral bioavailability and plasma lipid-lowering effect of probucol.

This work aimed to improve the oral bioavailability and plasma lipid-lowering effect of probucol (PB) by constructing a combined drug delivery system (CDDS) composed of nanostructured lipid carrier (NLC) and PEGylated poly(amidoamine) dendrimer (PEG-PAMAM). PEG-PAMAM with dendrimer generations of 5 (G5-PEG) or 7 (G7-PEG) were incorporated in PB-NLCs to form PB-CDDSs, PB-NLCs/G5-PEG and PB-NLCs/G7-PEG. The resultant two kinds of PB-CDDSs were characterized by particle size, zeta potential, drug encapsulation efficacy, PB release rates, and physical stability. Formulation effects of NLC and CDDS on the cellular uptake of hydrophobic drug were explored in Caco-2 cells by fluorescent Cy5 dye as a hydrophobic drug model. Furthermore, in vivo pharmacokinetics of the PB-CDDS composed of G5-PEG and PB-NLCs were investigated in a low density lipoprotein receptor knockout (LDLr-/-) mouse model, including plateau plasma PB concentrations after oral administration of multiple doses, and bioavailability after oral administration of a single dose of different PB formulations. In addition, lipid-lowering effect of PB-NLCs/G5-PEG was studied. The results indicate that both G5-PEG and G7-PEG significantly improved aqueous solubility of PB. The two PB-CDDSs exhibited similar particle size (around 150nm) as PB-NLCs, but slower PB burst release rate, higher total PB release amount, and better particle morphology and storage stability than PB-NLCs. In comparison with traditional NLC, CDDS dramatically enhanced cellular uptake of Cy5 into Caco-2 cells. In vivo results demonstrate that PB-NLCs/G5-PEG had the highest plateau plasma PB concentration and oral bioavailability, and the greatest cholesterol-lowering effect in comparison with PB suspensions and PB-NLCs. Therefore, G5-PEG incorporating NLC can be exploited as a promising drug delivery system to improve oral bioavailability and lipid-lowering effect of PB.