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Hydroxychloroquine (HCQ), the hydroxylated analog of chloroquine, is an antimalarial lysomotropic agent that inhibits autophagy due to lysosomal acidification, and subsequently blocks the fusion of autophagosomes with lysosomes which leads to the accumulation of autophagosomes that may accelerate tumor cell death. Given these hypothesis the aim of this study was to investigate the effects of HCQ in the inhibition of autophagy and the induction of apoptosis in cervical cancer SiHa cells. Cervical cancer SiHa cells were cultured with Hank's balanced salt solution (HBSS) as positive control of autophagy or treated with HCQ as part of the experimental groups. LC3 and P62/SQSTM1 were detected by quantitative polymerase chain reaction (qPCR) and western blotting, respectively in order to evaluate initially autophagosome formation and their degradation. Specific green fluorescent protein (GFP)-LC3 was subsequently detected by fluorescence microscopy in order to confirm the formation of autophagosomes. MTT and flow cytometry were adopted respectively to assess the proliferation and apoptosis of the SiHa cells. miRNA-9* was also investigated. The results demonstrated that HCQ increased the expressions of LC3 mRNA and LC3II protein and GFP-LC3 signalling but reduced the expression of p62/STSQM1 in cervical cancer SiHa cells. These results indicated HCQ has the ability to inhibit autophagy as incapable of degrading the autophagosome. However, HCQ may promote SiHa cell apoptosis as the MTT, apoptotic assay and miRNA-9* results revealed. HCQ has the ability to inhibit autophagy by blocking the degradation of autophagosomes and subsequently facilitates the apoptosis of cervical cancer SiHa cells.

[Display omitted] • Calcification was identified in Randall's plaque. • Calcification-related proteins were up-regulated in animal and cell models. • HCA reduces calcification-induced apoptosis and oxidative damage. • HCA inhibits cellular calcification and deposition of calcium salts in animals. Ectopic calcification of the renal interstitium as an important cause of renal stone formation. Hydroxycitric acid (HCA), a derivative of citric acid, has antioxidant properties and inhibits the crystallisation of calcium salts.The study found that HCA has an inhibitory effect on calcification and calcium salt deposition in fibroblasts. Additionally, HCA was found to reduce calcification-induced oxidative damage and apoptosis.Animal experiments also showed that HCA reduced calcium salt deposition and expression of calcification-related proteins. Cellular transcriptomic analysis demonstrated that HCA can regulate the changes in cellular transcriptome caused by calcification in culture. HCA can effectively inhibit the calcification of renal interstitial fibroblasts, reducing the risk of stone formation by inhibiting calcification culture-induced apoptosis and calcium salt deposition. Therefore, HCA is a promising alternative drug for inhibiting renal stone formation. [ABSTRACT FROM AUTHOR]

Abstract Perfluorooctanoic acid (PFOA) toxicity is of considerable concern due to its wide application, environmental persistence, and bioaccumulation. In the current study, we used a scaffold-free three-dimensional (3D) spheroid model of mouse liver cells (AML12) to explore the toxicity of PFOA and emerging alternatives (HFPO-DA and PFO4DA). Comparing the short-term (24 and 72 h treatment) toxicity of PFOA between conventional 2D monolayer cells and 3D spheroids, we found that spheroids had higher EC 50 values and lower ROS levels after treatment, indicating their greater resistance to PFOA. Cell viability (i.e., adenosine triphosphate (ATP) content and lactate dehydrogenase (LDH) leakage) and liver-specific function (i.e., albumin secretion) were stable in spheroids through 28 day of culture. However, under 100 and 200 μM-PFOA treatment for 28 day, ROS levels, LDH leakage, and caspase3/7 activity all increased significantly. As a sensitive parameter, ROS showed a significant increase at 21 day, even in the 50 μM-PFOA group. Consistent with the elevation of ROS and caspase3/7, the expressions of oxidative stress- and apoptosis-related genes, including Gsta2 , Nqo1 , Ho-1 , caspase3 , p53 , and p21 , were induced in dose- and time-dependent manners after PFOA exposure. The peroxisome proliferator-activated receptor alpha (PPARα) pathway was also activated after treatment, with significant induction of its target genes, Fabp4 and Scd1. Similar to PFOA, both HFPO-DA and PFO4DA activated the PPARα pathway, induced ROS levels, and initiated cell damage, though at a relatively lower extent than that of PFOA. Our results imply that the 3D spheroid model is a valuable tool in chronic toxicological studies. Graphical abstract Image 1 Highlights • PFOA toxicity differed between the 3D spheroid model and 2D monolayer cells. • ROS was a sensitive PFOA parameter in spheroids after long-term exposure. • PFOA activated the PPARα pathway in spheroids after long-term exposure. • HFPO-DA and PFO4DA hepatotoxicities were lower than that of PFOA. • The spheroid model is a feasible in vitro approach in chronic toxicological studies. [ABSTRACT FROM AUTHOR]

Background: Halofuginone, which is the main active ingredient of Dichroa fabrifuga, was used to inhibit the synthesis of type I collagen and played increasingly important roles in tumor therapy. This study aims to investigate the protective effects of halofuginone on human umbilical vein endothelial cells (HUVECs) from H2O2-induced apoptosis and oxidative stress. Methods: Propidium iodide and Annexin-V double staining assay was used to measure the apoptosis. Cell viability assay, the measurements of reactive oxygen species (ROS) parameters malondialdehyde and superoxide dismutase, western-blot assays, and quantitative PCR were used to elucidate the effects and mechanisms of halofuginone in protecting H2O2-induced injury. Results: The results showed that halofuginone counteracted H2O2-induced cell viability decline and PCNA downregulation. Furthermore, halofuginone decreased ROS levels and protected HUVECs from H2O2-induced apoptosis. In detail, it showed that H2O2 induced a transient activation of Mitogen-activated protein kinases members ERK1/2 and p38, whereas induced a sustained activation of c-Jun N-terminal kinase (JNK), which play dominant roles in triggering apoptosis. Inhibition of JNK activation also inhibited H2O2-mediated apoptosis. Finally, it was shown that halofuginone upregulated VEGF expressions, which functioned by inhibiting sustained JNK activation, thus protecting HUVECs. Conclusion: Halofuginone has powerful effects in protecting HUVECs from H2O2-induced apoptosis, via upregulating VEGF and inhibiting overactivated JNK phosphorylation. Halofuginone might be a promising preventive drug for cardiovascular diseases. [ABSTRACT FROM AUTHOR]

PURPOSE: To identify the incidence of cell apoptosis during premaxillary suture expansion. METHODS: Forty SD rats were randomly allotted into experimental and control groups. The experimental animals were subjected to premaxillary suture expansion by a helix spring with (200 ± 10)g initial force. They were sacrificed on day 1, 2, 3 and 5(n=5). The sequence of histological changes and osteoblast proliferation in the premaxillary suture were evaluated by hematoxylin and eosin(HE) and toluidine blue staining. Cellular apoptosis was identified by ssDNA immunostaining. The difference between experimental and control groups was analyzed by Student's t test with SPSS 15.0 software package. RESULTS: The width of premaxillary suture was enlarged 1 day after expansion and osteoblasts were accumulated along the suture. After 2 days of stretching, the number of the osteoblasts increased to the peak. New bone deposition was identified on day 3 and projection of suture with bone trabeculae was apparent on day 5. ssDNA immunostaining showed few apoptotic cells in premaxillary suture during natural growth. Upon expansion, cell apoptosis was identified in osteoblasts along the suture and the osteocytes embedded in premaxillary bone, especially on day 2 and day 3 (P<0.01). CONCLUSION: Apoptotic cell death might regulate the growth modification of premaxillary suture upon mechanical stretching. [ABSTRACT FROM AUTHOR]

The article presents a study which determines the effect of gelsolin, a calcium-regulated actin severing and capping protein, to the cardiac muscles. The study reveals that gelsolin has the ability to regulate cardiac modeling after myocardial infarction by DNase I-Mediated Apoptosis. Moreover, it also identified that gelsolin inhibition has the possibility to form a novel target for heart failure therapy.