Co-exposure of silica nanoparticles and methylmercury induced cardiac toxicity in vitro and in vivo.

The released nanoparticles into environment can potentially interact with pre-existing pollution, maybe causing higher toxicity. As such, assessment of their joint toxic effects is necessary. This study was to investigate the co-exposure cardiac toxicity of silica nanoparticles (SiNPs) and methylmercury (MeHg). Factorial design was used to determine the potential joint action type. In vitro study, human cardiomyocytes (AC16) were exposed to SiNPs and MeHg alone or the combination. Higher toxicity was observed on cell viability, cell membrane damage in co-exposure compared with single exposure and control. The co-exposure enhanced the ROS, MDA generation and reduced the activity of SOD and GSH-Px. In addition, the co-exposure induced much higher cellular apoptotic rate in AC16. In vivo study, after SD rats exposed to SiNPs and MeHg and their mixture by intratracheal instillation for 30days, pathological changes (myocardial interstitial edema) of heart were occurred in co-exposure compared with single exposure and control. Moreover obvious ultra-structural changes, including myofibril disorder, myocardial gap expansion, and mitochondrial damage were observed in co-exposure group. The activity of myocardial enzymes, including CK-MB, ANP, BNP and cTnT, were significantly elevated in co-exposure group of rat serum. Meanwhile, the cardiac injury-linked proteins expression showed an increase in SERCA2 and decreased levels of cTnT, ANP and BNP in co-exposure group. Factorial design analysis demonstrated that additive and synergistic interactions were responsible for the co-exposure cardiac toxicity in vitro and vivo. In summary, our results showed severe cardiac toxicity induced by co-exposure of SiNPs and MeHg in both cardiomycytes and heart. It will help to clarify the potential cardiovascular toxicity in regards to combined exposure pollutions.

Mitochondrial dysfunction, perturbations of mitochondrial dynamics and biogenesis involved in endothelial injury induced by silica nanoparticles.

As silica nanoparticles (SiNPs) pervade the global economy, however, the followed emissions during the manufacturing, use, and disposal stages inevitably bring an environmental release, potentially result in harmful impacts. Endothelial dysfunction precedes cardiovascular disease, and is often accompanied by mitochondrial impairment and dysfunction. We had reported endothelial dysfunction induced by SiNPs, however, the related mechanisms by which SiNPs interact with mitochondria are not well understood. In the present study, we examined SiNPs-induced mitochondrial dysfunction, and further demonstrated their adverse effects on mitochondrial dynamics and biogenesis in endothelial cells (HUVECs). Consequently, SiNPs entered mitochondria, caused mitochondrial swelling, cristae disruption and even disappearance. Further analyses revealed SiNPs increased the intracellular level of mitochondrial reactive oxygen species, eventually resulting in the collapse of mitochondrial membrane potential, impairments in ATP synthesis, cellular respiration and the activities of three ATP-dependent enzymes (including Na+/K+-ATPase, Ca2+-ATPase and Ca2+/Mg2+-ATPase), as well as an elevated intracellular calcium level. Furthermore, mitochondria in SiNPs-treated HUVECs displayed a fission phenotype. Accordingly, dysregulation of the key gene expressions (FIS1, DRP1, OPA1, Mfn1 and Mfn2) involved in fission/fusion event further certified the SiNPs-induced perturbation of mitochondrial dynamics. Meanwhile, SiNPs-treated HUVECs displayed declined levels of mitochondrial DNA copy number, PGC-1α, NRF1 and also TFAM, indicating an inhibition of mitochondrial biogenesis triggered by SiNPs via PGC-1α-NRF1-TFAM signaling. Overall, SiNPs triggered endothelial toxicity through mitochondria as target, including the induction of mitochondrial dysfunction, as well as the perturbations of their dynamics and biogenesis.

Metallothionein prevents doxorubicin cardiac toxicity by indirectly regulating the uncoupling proteins 2.

Doxorubicin (Dox) is a broad-spectrum anticancer agent, but its clinical use is restricted due to irreversible cardiac toxicity. Metallothionein (MT) can inhibit Dox-induced cardiac toxicity. Applying a proteomics approach we determined that uncoupling proteins (UCPs) may be implicated in this process. This study was designed to examine the mechanisms of MT against Dox cardiac toxicity and the link between MT and UCP2. In vivo, wild-type (MT+/+) and MT-I/II null (MT-/-) mice were given a single dose of Dox (15 mg/kg, i.p.) and sacrificed at 4 days after Dox injection. In vitro, cardiomyocytes were prepared from MT-/- and MT+/+ neonatal mice and cardiomyocytes were pretreated with typical antioxidant NAC or the UCP2 inhibitor genipin followed by exposure to Dox. Based on the results, genipin enhanced Dox-induced oxidative injury, particularly in MT-/- cardiomyocyte. UCP2 levels in MT-/- mice were significantly lower compared to MT+/+ mice treated with Dox. Co-immunoprecipitation demonstrated that MT did not directly bind to UCP2. The NAC and Nrf2 activator oltipraz inhibit the decrease of UCP2 expression induced by Dox. Therefore, attenuating free radical damage with UCP2 help MT antagonize the Dox-induced cardiac toxicity, but does not directly bind MT. MT may regulate UCP2 expression by up-regulating Nrf2.

Gene expression profiles and bioinformatics analysis of human umbilical vein endothelial cells exposed to PM2.5.

Cardiovascular system is demonstrated the main target of PM2.5 and the objective of this study was to explore the toxic effect and molecular mechanisms caused by PM2.5 in primary human umbilical vein endothelial cells (HUVECs) using microarray and bioinformatics analysis. The results showed that 591 genes were differentially expressed triggered by PM2.5, of which 174 genes were down-regulated, while 417 genes were up-regulated. Gene ontology analysis revealed that PM2.5 caused significant changes in gene expression patterns, including response to stimuli, immune response, and cellular processes. Pathway analysis and Signal-net analysis suggested that endocytosis, chemokine signaling pathway, RNA transport, protein processing in endoplasmic reticulum (ER) and autophagy regulation were the most critical pathways in PM2.5-induced toxicity in HUVECs. Moreover, gene expression confirmation of LIF, BCL2L1, CSF3, HMOX1, RPS6, PFKFB, CAPN1, HSPBP1, MOGS, PREB, TUBB2A, GABARAP by qRT-PCR indicated that endocytosis might be involved in the cellular uptake of PM2.5 by forming phagosomes, and subsequently inflammation, hypoxia and ER stress was occurred, which finally activated autophagy after PM2.5 exposure in HUVECs. In summary, our data can serve as fundamental research clues for further studies of PM2.5-induced toxicity in HUVECs.

1H NMR-based metabolomics study on repeat dose toxicity of fine particulate matter in rats after intratracheal instillation.

Systemic metabolic effects and toxicity mechanisms of ambient fine particulate matter (PM2.5) remain uncertain. In order to investigate the mechanisms in PM2.5 toxicity, we explored the endogenous metabolic changes and possible influenced metabolic pathways in rats after intratracheal instillation of PM2.5 by using a 1H nuclear magnetic resonance (NMR)-based metabolomics approach. Liver and kidney histopathology examinations were also performed. Chemical characterization demonstrated that PM2.5 was a complex mixture of elements. Histopathology showed cellular edema in liver and glomerulus atrophy of the PM2.5 treated rats. We systematically analyzed the metabolites changes of serum and urine in rats using 1H NMR techniques in combination with multivariate statistical analysis. Significantly reduced levels of lactate, alanine, dimethylglycine, creatine, glycine and histidine in serum, together with increased levels of citrate, arginine, hippurate, allantoin and decreased levels of allthreonine, lactate, alanine, acetate, succinate, trimethylamine, formate in urine were observed of PM2.5 treated rats. The mainly affected metabolic pathways by PM2.5 were glycine, serine and threonine metabolism, glyoxylate and dicarboxylate metabolism, citrate cycle (TCA cycle), nitrogen metabolism and methane metabolism. Our study provided important information on assessing the toxicity of PM2.5 and demonstrated that metabolomics approach can be employed as a tool to understand the toxicity mechanism of complicated environmental pollutants.

Macrophages participate in local and systemic inflammation induced by amorphous silica nanoparticles through intratracheal instillation.

Silica nanoparticles (SiNPs) are amongst the most commonly used materials in the field of nanomedicine and, therefore, their influence on organisms has drawn increasing attention in recent years. Most reports have focused on the single tissue reactions induced by SiNPs. Herein, the reaction of primary organs to SiNPs following intratracheal instillation in mice was analyzed by histopathology and ultrastructure observation. Following elucidation of the role of macrophages in local and systemic inflammation, the underlying mechanisms were explored using a macrophage cell line in vitro. The results suggest that macrophages swallow the SiNPs and secrete inflammatory factors by activating the NLRP3 inflammasome, thus participating in local and systemic inflammation.

[Study on the weight-reducing effect of Acer truncatum leave extract in alimentary obesity rat].

OBJECTIVE: To investigate the weight-reducing effect of Acer truncatum leave extract on alimentary obesity rats and its effect on fatty acid synthase (FAS). METHODS: SPF-grade adult male Wistar rats were fed with high-fat diet and Acer truncatum leave extract (10, 30 and 100 mg/kg BW) was given by gavage once a day for 31 days. Body weight (BW), adipose weight and food consumption were recorded, and the activity of hepatic fatty acid synthase (FAS) was measured. RESULTS: Compared with the model-control group, body weight, adipose weight and the ratio of adipose weight to body weight were obviously lower in 30 mg/kg BW and 100 mg/kg BW groups (P < 0.01). The activity of hepatic FAS in rats administrated with the extract was markedly lower than the control rats (P < 0.05). CONCLUSION: There might have a function of Acer truncatum leave extract on reducing body weight.

Weight-reducing effect of Acer truncatum Bunge may be related to the inhibition of fatty acid synthase.

In order to study the anti-obesity ability and inhibition towards fatty acid synthase (FAS) of the extract, a 70% ethanol extract of Acer truncatum Bunge (AT) leaves was further extracted with ethyl acetate. FAS is a very significant lipogenic enzyme, participating in energy metabolism in vivo; it has also been observed that FAS inhibitors might be potent anti-obesity agents. Experimental results on animals showed that the extract significantly reduced food intake and adipose, and effectively controlled weight evolution. Lipogenesis inhibition might be regarded as one of the reasons for the weight control and adipose reduction by AT. The extract was further isolated using a series of column chromatography that yielded 10 known compounds. 1,2,3,4,6-Penta-O-galloyl-ß-D-glucose was found to be one of the major active constituents in the extract of AT.

The novel inhibitory effect of Pangdahai on fatty acid synthase.

Recently, animal fatty acid synthase (FASN) is reported as a potential therapeutic target for obesity and cancer. Considerable interest has been developed in searching for novel inhibitors of this enzyme. An extract from Pangdahai has been found to inhibit FASN in both reversible and irreversible manners, with an IC(50) of 3.5 microg/ml and an apparent inactivation rate constant of k(obs) of 2.2 x 10(-3)/min. The kinetic study showed that the Pangdahai extract inhibited the overall FASN reaction uncompetitively with acetyl-CoA, but it presented in a mixed manner both with NADPH and with malonyl-CoA. Its major reacting site on this enzyme, as compared between two IC(50) values, is not in the beta-ketoacyl reduction domain. A weight reducing experiment in rats showed that the extract significantly reduced the adipose and food intake, but in view of statistics (P < 0.05), a correlation between the reductions in the adipose and in the food consumption and the inhibition of hepatic FASN could not be established. Three known flavonoid compounds were isolated from the extract and the structure-activity relationship was discussed.