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Porcine epidemic diarrhea virus (PEDV), a member of Coronaviridae, causes high mortality in newborn piglets, and has caused significant economic losses in the pig industry. PEDV infection can induce apoptosis, both caspase-dependent and caspase-independent, but the details of apoptosis remain clarified. This study investigated the effect of death receptor DR5 on PEDV infection and its relationship with PEDV-induced apoptosis. We found that DR5 knockdown reduced viral mRNA and protein levels of PEDV, and the viral titer decreased from 104.5 TCID50 to 103.4 TCID50 at 12 hpi. Overexpression of DR5 significantly increased the viral titer. Further studies showed that DR5 facilitates viral replication by regulating caspase-8-dependent apoptosis, and the knockdown of DR5 significantly reduced PEDV-induced apoptosis. Interestingly, we detected a biphasic upregulation expression of DR5 in both Vero cells and piglets in response to PEDV infection. We found that DR5 also facilitates viral entry of PEDV, especially, incubation with DR5 antibody can reduce the PEDV binding to Vero cells. Our study improves the understanding of the mechanism by which PEDV induces apoptosis and provides new insights into the biological function of DR5 in PEDV infection. [ABSTRACT FROM AUTHOR]

In the present study, a polysaccharide fraction (PTP) was isolated and purified from the tubers of Pinellia ternata. We researched its structure and anti-tumor activity, and further studied its molecular mechanism of inducing apoptosis of Hep G2 cells. The results indicated that PTP was an acid heteropolysaccharide and the average molecular weight of PTP identified by HPGPC was 3.06 × 106 Da. Ion chromatography (IC) determined that PTP was mainly composed of Ara:Gal:Glu:Man:GlcA:GalA in a molar ratio of 6.98:16.56:7.25:2.04:1:4.16. Combined with the results of FT-IR and NMR spectroscopy, it was found that PTP is a pyranose containing α-configuration and β-configuration, mainly consist of β-D-Gal, α-D-Glu, α-D-Ara and β-D-Man. By analyzing the results of MTT, cell cycle, Annexin V-FITC/PI double staining and cell morphology observation, we concluded that PTP induced dose-dependent apoptosis of Hep G2 cells via S phase arrest. In addition, mitochondrial membrane potential detection and Western blot further indicated that PTP was capable of inducing apoptosis in Hep G2 cells through an endogenous mitochondria-mediated apoptotic pathway. [ABSTRACT FROM AUTHOR]

Abstract Cardiac dysfunction is a vital complication during endotoxemia (ETM). Accumulating evidence suggests that enhanced glycolytic metabolism promotes inflammatory and myocardial diseases. In this study, we performed deep mRNA sequencing analysis on the hearts of control and lipopolysaccharide (LPS)-challenged mice (40 mg/kg, i.p.) and identified that the glycolytic enzyme, 6-phosphofructo-2-kinase (PFK-2)/fructose-2,6-bisphosphatase 3 (PFKFB3) might play an indispensable role in ETM-induced cardiac damage. Quantitative real-time PCR validated the transcriptional upregulation of PFKFB3 in the myocardium of LPS-challenged mice and immunoblotting and immunostaining assays confirmed that LPS stimulation markedly increased the expression of PFKFB3 at the protein level both in vivo and in vitro. The potent antagonist 3-(3pyridinyl)-1-(4-pyridinyl)-2-propen-1-one (3PO) was used to block PFKFB3 activity in vivo (50 mg/kg, i.p.) and in vitro (10 μM). Echocardiographic analysis and TUNEL staining showed that 3PO significantly alleviated LPS-induced cardiac dysfunction and apoptotic injury in vivo. 3PO also suppressed the LPS-induced secretion of tumor necrosis factor-α, interleukin (IL)-1β, IL-6 and lactate in the serum, in addition to lactate in the myocardium. PFKFB3 inhibition also diminished the nuclear translocation and phosphorylation of transcription factor nuclear factor-κB (NF-κB) in both adult cardiomyocytes and HL-1 cells. Furthermore, immunoblotting analysis showed that 3PO inhibited LPS-induced apoptotic induction in cardiomyocytes. Taken together, these findings demonstrate that PFKFB3 participates in LPS-induced cardiac dysfunction via mediating inflammatory and apoptotic signaling pathway. Highlights • The mRNA-seq data showed a vital role of PFKFB3 in LPS-caused cardiac dysfunction. • LPS stimulated significant protein upregulation of PFKFB3 in cardiomyocytes. • PFKFB3 blocker alleviated LPS-induced cardiac dysfunction and myocardial injury. • PFKFB3 may promote cardiac dysfunction via mediating inflammation and apoptosis. [ABSTRACT FROM AUTHOR]

[Display omitted] • Four undescribed seco -polyprenylated acylphloroglucinols from H. elodeoides. • A possible biosynthetic pathway of 1 – 4 from normal PAPs was proposed. • Compound 1 induced MCF-7 cell apoptosis. • Compound 1 arrested cell cycle at S phase via inducing oxidative DNA damage. Four undescribed seco -polyprenylated acylphloroglucinols (seco -PAPs), elodeoidesones A-D (1 – 4), were characterized from Hypericum elodeoides. Compound 1 represents the 1,6- seco -PAPs with fascinating 5/5 fused ring, while 2 – 4 possess a 1,2- seco -PAPs skeleton with a five-membered lactone core. Their structures including absolute configurations were established by spectroscopic analyses and quantum chemical computations. A possible biosynthetic pathway of 1 – 4 from normal PAPs was proposed. All the isolates were investigated for their cytotoxicity against tumor cells. Notably, 1 inhibited the proliferation of MCF-7 cells with the IC 50 value of 7.34 μM. Mechanism investigation indicated that 1 induced MCF-7 cells apoptosis by blocking cell cycle at S phase via inducing oxidative DNA damage. [ABSTRACT FROM AUTHOR]

Cardiac dysfunction is a vital complication of endotoxemia (ETM) with limited therapeutic options. Transient receptor potential canonical channel (TRPC)1 was involved in various heart diseases. While, the role of TRPC1 in ETM-induced cardiac dysfunction remains to be defined. In this study, we found that TRPC1 protein expression was significantly upregulated in hearts of lipopolysaccharide (LPS)-challenged mice. What's more, TRPC1 knockdown significantly alleviated LPS-induced cardiac dysfunction and injury. Further myocardial mRNA-sequencing analysis revealed that TRPC1 might participate in pathogenesis of ETM-induced cardiac dysfunction via mediating myocardial apoptosis and autophagy. Data showed that knockdown of TRPC1 significantly ameliorated LPS-induced myocardial apoptotic injury, cardiomyocytes autophagosome accumulation, and myocardial autophagic flux. Simultaneously, deletion of TRPC1 reversed LPS-induced molecular changes of apoptosis/autophagy signaling pathway in cardiomyocytes. Moreover, TRPC1 could promote LPS-triggered intracellular Ca2+ release, subsequent calpain activation and caveolin-1 degradation. Either blocking calpain by PD150606 or enhancing the amount of caveolin-1 scaffolding domain that interacts with TRPC1 by cell-permeable peptide cavtratin significantly alleviated the LPS-induced cardiac dysfunction and cardiomyocytes apoptosis/autophagy. Furthermore, cavtratin could inhibit LPS-induced calpain activation in cardiomyocytes. caveolin-1 could directly interact with calpain 2 both in vivo and in vitro. Importantly, cecal ligation and puncture-stimulated cardiac dysfunction and mortality were significantly alleviated in Trpc1 -/- and cavtratin-treated mice, which further validated the contribution of TRPC1-caveolin-1 signaling axis in sepsis-induced pathological process. Overall, this study indicated that TRPC1 could promote LPS-triggered intracellular Ca2+ release, mediate caveolin-1 reduction, and in turn activates calpain to regulate myocardial apoptosis and autophagy, contributing to ETM-induced cardiac dysfunction of mice. [Display omitted] • Deletion of TRPC1 alleviates LPS-induced cardiac dysfunction. • TRPC1 knockdown inhibits LPS-stimulated myocardial apoptosis and autophagy. • TRPC1 facilitates LPS-induced Ca2+ release and caveolin-1 degradation. • Direct protein interaction exists between caveolin-1 and calpain 2. • Targeting TRPC1-caveolin-1-calpain axis alleviates endotoxic cardiac dysfunction. [ABSTRACT FROM AUTHOR]

[Display omitted] • A pair of undescribed enantiomers (1a / 1b) was isolated from the roots of Ficus hirta. • Compound 4 showed enhanced cytotoxicity on HeLa, MCF-7 and H460 cells. • Compound 4 induced HeLa cells apoptosis and increased the PARP cleavage. • Compound 4 increased the expression of p-JNK and p-p38. • Compound 4 reduced the protein levels of p-AKT and p-ERK. A pair of undescribed enantiomers, (±) ficusflavonid A (1a / 1b), along with five known analogues, were isolated from the roots of Ficus hirta. Their structures were determined by the analysis of extensive spectroscopic data (including UV, IR, HRESIMS and NMR). Two enantiomers (1a and 1b) were successfully separated by chiral chromatographic column and their absolute configurations were assigned by the comparison of experimental and calculated ECD data. The cytotoxicity of all the isolates against HeLa, MCF-7, HepG2 and H460 cell lines were evaluated by MTT assay. Among them, 4 suppressed the proliferation of HeLa cells with the IC 50 value of 28.88 μM. Furthermore, the apoptotic effect of 4 on HeLa cells and the level of several crucial proteins in AKT/MAPKs signaling pathways were analyzed by flow cytometer and western blot assay. As a result, 4 induced HeLa cell apoptosis in a dose dependent manner and significantly increased the protein levels of p-JNK and p-p38, whereas distinctly reduced the expression of p-AKT, and p-ERK. Thus, compound 4 might induce HeLa cells apoptosis via MAPK and AKT signaling pathways, which could be considered as a potential leading compound for the development of anticancer drugs. [ABSTRACT FROM AUTHOR]

Myocardial ischemia/reperfusion (IR) injury is caused by the restoration of the coronary blood flow following an ischemic episode. Accumulating evidence suggests that galectin-3, a β-galactoside-binding lectin, acts as a biomarker in heart disease. However, it remains unclear whether manipulating galectin-3 affects the susceptibility of the heart to IR injury. In this study, RNA sequencing (RNA-seq) analysis identified that Lgals3 (galecin-3) plays an indispensable role in IR-induced cardiac damage. Immunostaining and immunoblot assays confirmed that the expression of galectin-3 was markedly increased in myocardial IR injury both in vivo and in vitro. Echocardiographic analysis showed that cardiac dysfunction in experimental IR injury was significantly attenuated by galectin-3 inhibitors including pectin (1%, i.p.) from citrus and binding peptide G3-C12 (5.0 mg/kg, i.p.). Galectin-3 inhibitor-treated mice exhibited smaller infarct sizes and decreased tissue injury. Furthermore, TUNEL staining showed that galectin-3 inhibition suppressed IR-mediated cardiomyocyte apoptosis. Mitochondrial membrane potential (MMP) and mitochondrial permeability transition pore (mPTP) levels were well-preserved and IR-induced changes of mitochondrial cyto c, cytosol cyto c, caspase-9, caspase-3, Bcl-2 and Bax in the galectin-3 inhibitor-treated groups were observed. Our findings indicate that the pathological upregulation of galectin-3 contributes to IR-induced cardiac dysfunction and that galectin-3 inhibition ameliorates myocardial injury, highlighting its therapeutic potential. [ABSTRACT FROM AUTHOR]

Background: Patatin-like phospholipase domain containing 5 (PNPLA5) is a newly-discovered lipase. Although the PNPLA family plays critical roles in diverse biological processes, the biological functions of PNPLA5 mostly unknown. We previously found that the deletion of Pnpla5 in rats causes a variety of phenotypic abnormalities. In this study, we further explored the effects of Pnpla5 knockout (KO) on male rats. Results: The body weight and testicular or epididymal tissue weight of three to six 3-month-old Pnpla5 KO or wild-type (WT) male Sprague–Dawley rats were measured. The protein expression levels were also measured via western blotting and iTRAQ (isobaric tags for relative and absolute quantitation) analyses. No significant difference between Pnpla5 KO and WT rats, regarding body weight, testicular or epididymal tissue weight, or hormone levels, were found. However, the relative testicular tissue weight of the KO (Pnpla5−/−) rats was higher (P < 0.05) than that of WT rats. Significant increases in apoptotic cells numbers (P < 0.001) and BAX and Caspase-9 expression levels were observed in the testicular tissue of Pnpla5−/− rats. Moreover, iTRAQ analysis revealed that the levels of proteins involved in steroid metabolism and wound healing were significantly decreased in Pnpla5−/− rats. Conclusion: This study revealed that Pnpla5 knockout induced apoptosis in rat testes. We also ascertained that Pnpla5 plays an important role in lipid metabolism, wound healing, and affects reproductive organs negatively, providing new target genes and pathways that can be analyzed to unravel the biological function of Pnpla5. [ABSTRACT FROM AUTHOR]

Deguelin is a rotenoid of the flavonoid family, which can be extracted from Lonchocarpus, Derris, or Tephrosia. It possesses the inhibition of cancer cell proliferation by inducing apoptosis through regulating the phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) signaling pathway, the NF-κB signaling pathway, the Wnt signaling pathway, the adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) signaling pathway and epidermal growth factor receptor (EGFR) signaling, activating the p38 mitogen-activated protein kinase (MAPK) pathway, repression of Bmi1, targeting cyclooxygenase-2 (COX-2), targeting galectin-1, promotion of glycogen synthase kinase-3β (GSK3β)/FBW7-mediated Mcl-1 destabilization and targeting mitochondria via down-regulating Hexokinases II-mediated glycolysis, PUMA-mediation, which are some crucial molecules which modulate closely cancer cell growth and metastasis. Deguelin inhibits tumor cell propagation and malignant transformation through targeting angiogenesis, targeting lymphangiogenesis, targeting focal adhesion kinase (FAK), inhibiting the CtsZ/FAK signaling pathway, targeting epithelial–mesenchymal transition (EMT), the NF-κB signaling pathway, regulating NIMA-related kinase 2 (NEK2). In addition, deguelin possesses other biological activities, such as targeting cell cycle arrest, modulation of autophagy, inhibition of hedgehog pathway, inducing differentiation of mutated NPM1 acute myeloid leukemia etc. Therefore, deguelin is a promising chemopreventive agent for cancer therapy. Anticancer activity of deguelin and its derivatives: recent advances in mechanisms of action (AMPK: Adenosine 5'-monophosphate (AMP)-activated protein kinase; COX-2: Cyclooxygenase; EGFR: Epidermal growth factor receptor; EMT: Epithelial–mesenchymal transition; FAK: Focal adhesion kinase; GSK3β: Glycogen synthase kinase-3; Hh: Hedgehog; MAPK: p38 mitogen-activated protein kinase; NF-κB: Nuclear factor κB; PI3K/Akt: Phosphoinositide 3-kinase/protein kinase B). [Display omitted] [ABSTRACT FROM AUTHOR]

Hematoporphyrin monomethyl ether (HMME) is a novel and promising porphyrin-related photosensitizer for photodynamic therapy (PDT). The aim of this study was to investigate HMME-induced apoptosis in CHMm cells, a canine breast cancer cell line. CHMm cells were treated with HMME and a He-Ne laser at a wavelength of 632.8 nm. Cell viability was determined using the trypan blue exclusion assay. Apoptosis was analyzed using Hoechst 33258, AO/EB, Annexin V/PI staining and single-cell gel electrophoresis (comet assay). Apoptotic morphology was further confirmed by Giemsa staining and transmission electron microscopy. Rates of apoptosis increased following PDT-HMME treatment in a time-dependent manner. Taken together, these results demonstrated that apoptosis plays a major role in PDT-HMME induced reduction in the viability of CHMm cells. [ABSTRACT FROM AUTHOR]

Abstract: B cell antigen receptor (BCR) density plays a role in the differentiation of immature B cells to their mature compartments; however, the exact strategy of its influence on the development of natural autoreactive B cells is still unclear. In the present study, we explored the role of BCR surface density in autoreactive B cell development by studying two lines of mice containing distinct copy numbers of an IgH transgene with VH derived from a poly-reactive natural antibody 3B4. Surface BCR levels were found to be related to the transgene copy number in these mice. In mice with higher copy numbers of the transgene, the BCRs were found to promote the remaining of autoreactive B cells into marginal zone (MZ) and B-1a subsets; meanwhile, elevated surface BCR levels were correlated with a significant decrease of follicular (Fo) B cell numbers and a reduction in the number of multiple stages of immature B cells both in spleen and bone marrow (BM). Interestingly, no difference in the ratio of cell apoptosis and proliferation was found in all stages of B cell development between two lines, except that more severely aberrant proliferation of pro/pre-B cells in BM was found in mice with higher transgene copies. This data supports the idea that natural poly-reactive B cells can be positively selected into MZ and B-1 cells, and high BCR surface density favors this selection. More importantly, our data suggests that the influence by receptor expression on the differentiation of natural poly-reactive B cells begins at an early stage of B cell development. [Copyright &y& Elsevier]

The expression of genes related to the Toll-like receptors (TLRs) signaling pathway were determined. Group A, B and C fed with basal diet and group D, E and F induced TD by feeding a basal diet containing 100 mg·kg−1 thiram. rGSTA3 protein was injected at 20 μg·kg−1 in group B, E and at 50 μg·kg−1 in C, F. Results suggested that lameness and death of chondrocytes were significant on day 14. TLRs signaling pathway related genes were screened based on the transcriptome enrichment, and validated on qPCR. IL-7, TLR2, 3, 4, 5, 7, 15, MyD88, MHC-II, MDA5 and TRAF6 were significantly (p < 0.05) expressed in group E and F as compared to group D on day 14 and 23. IL-7, MHCII, TRAF6, TLR3, TLR5, TLR7, and TLR15 determined insignificant in group D compared to group A on day 23. TD occur in an early phase and alleviated in the later period. rGSTA3 protein can prevent apoptosis and repair degraded chondrocytes. Image 1 • TD is an important undetected avian growth plate disease. • Immune genes regulate apoptosis, ultimately death of chondrocytes occured. • Toll-like receptor (TLRs) mediates cellular immune responses to cause apoptosis. • rGSTA3 protein treatment prevents the apoptosis by downregulating TLRs on day 14 and day 23. [ABSTRACT FROM AUTHOR]

Two new compounds, triacremoniate (1) and dietziamide C (2) along with known compounds β-Adenosine (3) and acrepyrone A (4) were obtained from the mangrove-derived fungus Acremonium citrinum. MMF4. Their structures were unambiguously determined by extensive spectroscopic methods, including UV, IR, HRESIMS and NMR. Triacremoniate (1) can promote apoptosis of HeLa cells by increasing the PARP cleavage and the phosphorylation of JNK and p38. Unlabelled Image • Two new compounds were obtained from the mangrove-derived fungus Acremonium citrinum. • Compound 1 showed obvious cytotoxicity against Hela cell line. • Compound 1 distinctly induced the apoptosis of Hela cell line. • Compound 1 significantly increased the expression of p-JNK and p-p38 in MAPK signaling pathway. [ABSTRACT FROM AUTHOR]