Macranthoidin B (MB) Promotes Oxidative Stress-Induced Inhibiting of Hepa1-6 Cell Proliferation via Selenoprotein.

The aim of this study was to investigate the role of selenoproteins in Macranthoidin B (MB) with regard to the inhibition of hepa1-6 cell proliferation. The CCK8 method was used to detect the inhibition rate in hepa1-6 cell of proliferation. The production of ROS, MDA, GSH levels, and GSH-Px and SOD activities was detected according to corresponding reagent kits. We determined the mRNA expressions of 25 selenoproteins in hepa1-6 cells via real-time quantitative PCR (qRT-PCR); moreover, the heat map and principal component analysis were used for further bioinformatics analysis. The results revealed that with an increasing concentration of MB, the inhibitory effect on hepa1-6 cell proliferation intensified. Compared with the control group, the treatment group showed significantly increased ROS levels, elevated MDA contents, and decreased GSH level, GSH-Px activity, and SOD activity. Increasing MB concentration treatment induced remarkable degradation of Txnrd1, Txnrd2, Txnrd3, Gpx1, Gpx2, Gpx3, Gpx6, Dio1, Dio2, Selt, Selp, Selh, Selk, Selw, Seln, and Dio3. Principal component analysis revealed that Txnrd 3, Selk, Selo, Selw, Selt, Dio2, Txnrd1, Dio3, Gpx6, and Dio1 were highly correlated with MB. In conclusion, MB dose dependently inhibited hepa1-6 cell proliferation and induced oxidative stress. Based on bioinformatics analysis, with MB treatment, Txnrd 3, Selk, Selo, Selw, Selt, Dio2, Txnrd1, Dio3, Gpx6, and Dio1 exhibited critical role in the inhibition of hepa1-6 cells proliferation. The functions of these selenoproteins were associated with oxidative stress.

Schisandrin B Induced ROS-Mediated Autophagy and Th1/Th2 Imbalance via Selenoproteins in Hepa1-6 Cells.

Schisandrin B (Sch B) is well-known for its antitumor effect; however, its underlying mechanism remains confusing. Our study aimed to investigate the role of selenoproteins in Sch B-induced autophagy and Th1/Th2 imbalance in Hepa1-6 cells. Hepa1-6 cells were chosen to explore the antitumor mechanism and were treated with 0, 25, 50, and 100 µM of Sch B for 24 h, respectively. We detected the inhibition rate of proliferation, transmission electron microscopy (TEM), monodansylcadaverine (MDC) staining, reactive oxygen species (ROS) level and oxidative stress-related indicators, autophagy-related genes, related Th1/Th2 cytokines, and selenoprotein mRNA expression. Moreover, the heat map, principal component analysis (PCA), and correlation analysis were used for further bioinformatics analysis. The results revealed that Sch B exhibited well-inhibited effects on Hepa1-6 cells. Subsequently, under Sch B treatment, typical autophagy characteristics were increasingly apparent, and the level of punctate MDC staining enhanced and regulated the autophagy-related genes. Overall, Sch B induced autophagy in Hepa1-6 cells. In addition, Sch B-promoted ROS accumulation eventually triggered autophagy initiation. Results of Th1 and Th2 cytokine mRNA expression indicated that Th1/Th2 immune imbalance was observed by Sch B treatment in Hepa1-6 cells. Intriguingly, Sch B downregulated the majority of selenoprotein expression. Also, the heat map results observed significant variation of autophagy-related genes, related Th1/Th2 cytokines, and selenoprotein expression in response to Sch B treatment. PCA outcome suggested the key role of Txnrd1, Txnrd3, Selp, GPX2, Dio3, and Selr with its potential interactions in ROS-mediated autophagy and Th1/Th2 imbalance of Hepa1-6 cells. In conclusion, Sch B induced ROS-mediated autophagy and Th1/Th2 imbalance in Hepa1-6 cells. More importantly, the majority of selenoproteins were intimately involved in the process of autophagy and Th1/Th2 imbalance, Txnrd3, Selp, GPX2, Dio3, and Selr had considerable impacts on the process.

Selenium-mediated gga-miR-29a-3p regulates LMH cell proliferation, invasion, and migration by targeting COL4A2.

Selenium (Se) is an essential trace element that has several functions in cellular processes related to cancer prevention. While the cancericidal effect of Se has been reported in liver cancer, the mechanism has not been clarified. MiR-29a has widely been reported as a tumor suppressor; however, it also acts as a carcinogenic agent by increasing cell invasion in human epithelial cancer cells and hepatoma cells. In a previous study, we found that miR-29a-3p is a Se-sensitive miRNA. However, its effect in the chicken hepatocellular carcinoma cell line (LMH) is still unknown. In the present study, we found that the expression of miR-29a-3p in LMH cells was decreased by Se supplementation and increased under Se-deficient conditions. Flow cytometry and CCK-8 results suggested that Se decreased LMH cell proliferation induced by miR-29a-3p overexpression. Transwell and gap-closure assays implied that Se mediated LMH cell invasion and migration by downregulating miR-29a-3p. Quantitative real-time polymerase chain reaction and Western blotting results suggested that Se mitigated miR-29a-3p overexpression-induced LMH cell proliferation by downregulating CDK2, cyclin-D1, CDK6, and cyclin-E1. We further demonstrated that collagen type IV alpha 2 (COL4A2) is a target gene of miR-29a-3p. COL4A2 activates the RhoA/ROCK pathway to promote LMH cell invasion and migration. In conclusion, Se mediated miR-29a-3p overexpression induced LMH cell invasion and migration by targeting COL4A2 to inactivate the RhoA/ROCK pathway.

The Effectiveness of Different Doses of Intravenous Immunoglobulin on Severe Hand, Foot and Mouth Disease: A Meta-Analysis.

OBJECTIVE: To conduct a meta-analysis of evidence from randomized controlled trails (RCTs) of different doses of intravenous immunoglobulin (IVIG) in children with severe hand, foot and mouth disease (HFMD) to provide the scientific basis for clinical practice. METHODS: A search of PubMed-Medline, CNKI, Wanfang, and VIP database (until June 30, 2017) was performed and Software RevMan5.3 was used to evaluate the effect of different doses of IVIG on HFMD in RCTs. We used random-effects models (or fixed-effects models) and generic inverse variance methods to process quantitative data, followed by a leave-one-out method for sensitivity analysis. RESULTS: From a total of 420 entries identified via searches, 8 RCTs involving 1,450 patients were included in the final analysis. The results of the meta-analysis showed that compared with conventional therapy alone, conventional therapy combined with IVIG had shorter fever clearance time, shorter rash regression time, and shorter clinical cure time. Subgroup analyses showed that the high-dose group (1 g/kg/day) had shorter fever clearance time (p < 0.05), shorter rash regression (p< 0.05), shorter remission time of neurological symptoms (p < 0.05), but longer clinical cure time (p > 0.05). CONCLUSION: The high-dose group has a better prognosis; however, the advantages and disadvantages should be carefully considered when deciding the doses in the treatment of severe HFMD.

MicroRNA-193b-3p regulates hepatocyte apoptosis in selenium-deficient broilers by targeting MAML1.

Selenium (Se) is an important nutritional element in the diet. Apoptosis is one of the characteristic pathological changes in liver tissue resulting from Se deficiency. MicroRNA (miRNA) plays an important role in cell proliferation, differentiation, apoptosis and tumorigenesis. However, why apoptosis occurs during Se deficiency and how miRNA regulates hepatocyte apoptosis in broilers requires further study. We used a dual luciferase reporter assay system and quantitative real-time PCR (qPCR) to screen hepatocytes in Se-deficient broilers for the specificity of hepatocyte apoptosis miRNA and its target protein. We tested the apoptosis of Se-deficient broiler livers and microRNA-193b-transfected primary hepatocytes using qPCR, western blot (WB) and flow cytometry. Our studies revealed that Se deficiency led to microRNA-193b-3p (miR-193b-3p) overexpression and increased apoptosis-related gene expression, resulting in broiler hepatocyte apoptosis. Mastermind-like protein 1 (MAML1) was one of the miR-193b-3p targets, and its expression was down-regulated in miR-193b-3p-overexpressing hepatocytes. Further studies have shown that miR-193b-3p overexpression induced changes of apoptosis-related gene expression by inhibiting the release of MAML1. Interestingly, when we overexpressed miR-193b-3p, which was added to the Signal transducers and activators of transcription-1 (STAT1) inhibitor fludarabine (Flu), hepatocyte apoptosis was significantly reduced. When these results were combined, they indicated that miR-193b-3p is involved in broiler hepatocyte apoptosis in Se deficiency by regulating the target protein MAML1. This finding may provide new ideas for studying the mechanism of hepatocyte injury due to Se deficiency.

Lower Selenoprotein T Expression and Immune Response in the Immune Organs of Broilers with Exudative Diathesis Due to Selenium Deficiency.

The objective of the present study was to investigate whether dietary selenium (Se) deficiency would affect the expression of selenoprotein T (SelT) and immune response in the immune organs of broilers. Changes in expression of inflammatory cytokines and oxidative stress response caused by Se deficiency can lead to organism damage, which in turn leads to immune response. Sixty (1-day-old) broilers were divided into the control group and Se-deficiency group. Animal models with exudative diathesis were duplicated in the broilers by feeding them Se-deficient diet for 20 days. After the Se-deficient group exhibited symptoms of exudative diathesis, all the broilers were euthanized, and their immune organs were taken for analysis. The tissues including spleen, bursa of Fabricius, and thymus were treated to determine the pathological changes (including microscopic and ultramicroscopic), the messenger RNA (mRNA) expression levels of SelT and its synthetase (SecS and SPS1), cytokine mRNA expression levels, and antioxidant status. The microscopic and ultramicroscopic analyses showed that immune tissues were obviously injured in the Se-deficient group. The mRNA expression of SelT was decreased compared with that in the control group. Meanwhile, the mRNA expression levels of SecS and SPS1 were downregulated. In the Se-deficient group, the mRNA expression levels of IL-1R and IL-1ß were higher than those of three control organs. Additionally, the IL-2 and INF-γ mRNA expression levels were lower than those of the control group. The activity of CAT was decreased, and the contents of H2O2 and •OH were increased due to Se deficiency. Pearson method analysis showed that the expression of SelT had a positive correlation with IL-2, INF-γ, SecS, and SPS1 and a negative correlation with IL-1R and IL-1ß. In summary, these data indicated that Se-deficient diet decreased the SelT expression and its regulation of oxidative stress, and it inhibited a pleiotropic mechanism of the immune response.

Involvement of mitochondrial pathway in environmental metal pollutant lead-induced apoptosis of chicken liver: perspectives from oxidative stress and energy metabolism.

This study aimed to investigate the possible mechanisms of environmental metal pollutant lead (Pb)-induced apoptosis in chicken. Forty 8-day-old healthy chickens were randomly assigned to two groups (n = 20/group) after raising standard commercial diet and drinking water for 1 week: including control group and Pb group ((CH3COO)2Pb 350 mg/L of drinking water); the chickens were given euthanasia and collected livers at 90 days. A significant increase of apoptosis rate were found in Pb group and Pb induced obvious ultrastructural changes of chicken liver. The mRNA levels of glycometabolism key enzymes were significantly lower in Pb group than those in controls. Higher levels of malondialdehyde (MDA) and nitric oxide (NO) were observed in Pb group; the activities of antioxidant enzymes and ATPases were significantly lower in Pb group than those in controls, while the inducible nitric oxide synthase (iNOS) activity was on the contrary. The mRNA and protein levels of pro-apoptotic genes were all lower in Pb group than those in controls. Altogether, Pb-induced mitochondrial swelling and nuclear chromatin condensation, oxidative stress, energy metabolism disorder, thereby lead to apoptosis via mitochondrial pathway in chicken liver, suggesting that Pb-induced mitochondrial pathway apoptosis plays an important role in the mechanisms of Pb cytotoxicity to chicken liver.

Alleviation Mechanisms of Selenium on Cadmium-Spiked Neutrophil Injury to Chicken.

To determine the negative effects of cadmium (Cd) exposure and the protective role of selenium (Se) on Cd-spiked neutrophils of chicken, forty-eight 28-day-old Isa Brown male chickens were divided randomly into four groups. Group I (control group) was fed with the basic diet containing 0.2 mg/kg Se. Group II (Se-treated group) was fed with the basic diet supplemented with Na2SeO3, and the total Se content was 2 mg/kg. Group III (Se/Cd-treated group) was fed with the basic diet supplemented with Na2SeO3; the total Se content was 2 mg/kg and supplemented with 150 mg/kg CdCl2. Group IV (Cd-treated group) was fed with the basic diet supplemented with 150 mg/kg CdCl2. Analyses of inflammatory factors, cytokines, and heat shock protein (Hsp) messenger RNA (mRNA) expression were detected by real-time PCR (RT-PCR). Additionally, we evaluated the phagocytic rate of neutrophils in peripheral blood. First, we observed that Cd significantly induced the mRNA expression levels of inflammatory factors NF-κB, iNOS, COX-2, and TNF-α, while Se/Cd treatment reduced their mRNA expression, although these expression levels remained higher than that of the control group. In addition, the mRNA expression levels of cytokines (IL-2, IL-4, and IL-10) for the Se-treated group exhibited significant differences between the Se/Cd-treated group and the Cd-treated group. Furthermore, the mRNA expression levels of Hsps demonstrated that the Se/Cd-treated group and the Cd-treated group were significantly higher (P < 0.05) than the control group and the Se-treated group. These results demonstrated that Se presented partial protection on Cd-spiked neutrophils of chicken with Hsps being involved in the process of the Cd-spiked toxic effects in chicken peripheral blood neutrophils.

Possible correlation of selenoprotein W with inflammation factors in chicken skeletal muscles.

The aim of the present study was to investigate the possible correlation of selenoprotein W (SelW) with inflammatory injury induced by dietary selenium (Se) deficiency in chicken. One-day-old male chickens were fed either a commercial diet or a Se-deficient diet for 55 days. Then, the expression levels of SelW messenger RNA (mRNA) and inflammation-related genes (NF-κB, TNF-α, iNOS, COX-2, and PTGES) in chicken skeletal muscles (wing muscle, pectoral muscle, and thigh muscle) were determined at 15, 25, 35, 45, and 55 days old, respectively. In addition, the correlation between SelW mRNA expression and inflammation-related genes were assessed. The results showed that dietary Se deficiency reduced the mRNA expression of SelW in chicken wing, pectorals, and thigh muscles. In contrast, Se deficiency increased the mRNA expression levels of inflammation-related genes in chicken skeletal muscle tissues at different time points. The Pearson's correlation coefficients showed that the mRNA expression levels of inflammation-related genes were significantly negative related to SelW (p < 0.05). These data showed that Se deficiency induced the inflammatory response in chicken skeletal muscle. As one important selenoprotein gene in skeletal muscles, SelW may play a role in the regulation of inflammation reaction in Se-deficiency myopathy.

Selenium deficiency inhibits the conversion of thyroidal thyroxine (T4) to triiodothyronine (T3) in chicken thyroids.

Selenium (Se) influences the metabolism of thyroid hormones in mammals. However, the role of Se deficiency in the regulation of thyroid hormones in chickens is not well known. In the present study, we examined the levels of thyroidal triiodothyronine (T3), thyroidal thyroxine (T4), free triiodothyronine, free thyroxine (FT4), and thyroid-stimulating hormone in the serum and the mRNA expression levels of 25 selenoproteins in chicken thyroids. Then, principal component analysis (PCA) was performed to analyze the relationships between the selenoproteins. The results indicated that Se deficiency influenced the conversion of T4 to T3 and induced the accumulation of T4 and FT4. In addition, the mRNA expression levels of the selenoproteins were generally decreased by Se deficiency. The PCA showed that eight selenoproteins (deiodinase 1 (Dio1), Dio2, Dio3, thioredoxin reductase 2 (Txnrd2), selenoprotein i (Seli), selenoprotein u (Selu), glutathione peroxidase 1 (Gpx1), and Gpx2) have similar trends, which indicated that they may play similar roles in the metabolism of thyroid hormones. The results showed that Se deficiency inhibited the conversion of T4 to T3 and decreased the levels of the crucial metabolic enzymes of the thyroid hormones, Dio1, Dio2, and Dio3, in chickens. In addition, the decreased selenoproteins (Dio1, Dio2, Dio3, Txnrd2, Seli, Selu, Gpx1, and Gpx2) induced by Se deficiency may indirectly limit the conversion of T4 to T3 in chicken thyroids. The information presented in this study is helpful to understand the role of Se in the thyroid function of chickens.