Acacetin reduces intrasynaptosomal Ca²⁺ levels but does not alter the synaptosomal membrane potential and Na⁺ influx.

<p>Cytosolic free Ca²⁺ concentration (nM) (A) or synaptosomal membrane potential (B) was measured in the absence (control) and in the presence of 30 µM acacetin, added 10 min before depolarization with 1 mM 4-AP. C: Na⁺ influx was induced by 1 mM 4-AP in the absence (control) or presence of 30 µM acacetin or 2 µM TTX, added 10 min before depolarization. Results are mean ± SEM of independent experiments, using synaptosomal preparations from five animals. ***, <i>P</i><0.001 versus control group.</p

Acacetin suppresses KA-induced microglial activation in the CA3 region of the hippocampus.

<p>Acacetin (10 or 50 mg/kg, i.p.) was administrated 30 min before KA injection, and the hippocampal sections were stained with anti-OX-42 antibody at 3 days after KA injection. Representative photomicrographs illustrating OX-42 immunoreactivity in the hippocampal CA3 region of control, KA, KA + acacetin 10 mg/kg, and KA + acacetin 50 mg/kg. Insets in figure show the morphological changes after KA administration under higher magnification. Representative picture from six independent experiments were presented. Scale bar  = 100 µm for A-D.</p

Acacetin attenuates KA-induced neuronal cell death in the CA3 region of the hippocampus.

<p>Acacetin (10 or 50 mg/kg) was administrated intraperitoneally 30 min before KA injection, and extents of neuronal losses in the hippocampus were evaluated at 3 days after KA injection by staining with neutral red (A-D) and Fluoro-Jade B (E-H). Representative photomicrographs illustrating neuronal cell death in the hippocampal CA3 region of control, KA, KA + acacetin 10 mg/kg, and KA + acacetin 50 mg/kg. (I) Quantification of Fluoro-Jade B-positive neurons in the CA3 region of the hippocampus. Data are expressed as mean ± SEM of six independent experiments. ***<i>P</i><0.001, as compared with the KA-treated group. Scale bar for A-D, E-H = 250 µm, A′-D′ = 100 µm.</p

Neurocytoprotective Effects of Aliphatic Hydroxamates from Lovastatin, a Secondary Metabolite from <i>Monascus</i>-Fermented Red Mold Rice, in 6‑Hydroxydopamine (6-OHDA)-Treated Nerve Growth Factor (NGF)-Differentiated PC12 Cells

Lovastatin, a secondary metabolite isolated from <i>Monascus</i>-fermented red rice mold, has neuroprotective activity and permeates the blood–brain barrier. The aim of this study was to enhance the activity of lovastatin for potential use as a treatment for neuronal degeneration in Parkinson’s disease. Six lovastatin-derived compounds were semisynthesized and screened for neurocytoprotective activity against 6-hydroxydopamine (6-OHDA)-induced toxicity in human neuroblastoma PC12 cells. Four compounds, designated as <b>3a</b>, <b>3d</b>, <b>3e</b>, and <b>3f</b>, significantly enhanced cell viability. In particular, compound <b>3f</b> showed excellent neurocytoprotective activity (97.0 ± 2.7%). Annexin V-FITC and propidium iodide double staining and 4′,6-diamidino-2-phenylindole staining indicated that compound <b>3f</b> reduced 6-OHDA-induced apoptosis in PC12 cells. Compound <b>3f</b> also reduced caspase-3, -8, and -9 activities, and intracellular calcium concentrations elevated by 6-OHDA in a concentration-dependent manner, without inhibiting reactive oxygen species generation. JC-1 staining indicated that compound <b>3f</b> also stabilized mitochondrial membrane potential. Thus, compound <b>3f</b> may be used as a neurocytoprotective agent. Future studies should investigate its potential application as a treatment for Parkinson’s disease

Total Synthesis of Hispidulin and the Structural Basis for Its Inhibition of Proto-oncogene Kinase Pim‑1

A new method is applied to synthesize hispidulin, a natural flavone with a broad spectrum of biological activities. Hispidulin exhibits inhibitory activity against the oncogenic protein kinase Pim-1. Crystallographic analysis of Pim-1 bound to hispidulin reveals a binding mode distinct from that of quercetin, suggesting that the binding potency of flavonoids is determined by their hydrogen-bonding interactions with the hinge region of the kinase. Overall, this work may facilitate construction of a library of hispidulin-derived compounds for investigating the structure–activity relationship of flavone-based Pim-1 inhibitors

<i>N</i>‑Hydroxycinnamide Derivatives of Osthole Presenting Genotoxicity and Cytotoxicity against Human Colon Adenocarcinoma Cells in Vitro and in Vivo

Osthole is extracted from the Chinese herbs <i>Cnidium monnieri</i> and <i>Angelica pubescens</i>, and it was found to have antitumor activity in vitro and in vivo. A series of osthole derivatives have been synthesized, and the <i>N</i>-hydroxycinnamide derivatives of osthole, WJ1376-1 and WJ1398-1 were found to have the greatest potential against human colon adenocarcinoma cells. In contrast to the parental osthole, both WJ1376-1 and WJ1398-1 were found to induce multinucleation and polyploidy by microscopic observation and flow cytometry. WJ1376-1 and WJ1398-1 significantly activated ataxia telangiectasia and rad3 related (ATR) kinase, which triggered activation of the checkpoint kinase 2 (Chk2) signaling pathway and then down regulated Cdc25 phosphatase and Cdc2/cyclin B kinase activities. WJ1376-1 and WJ1398-1 also inhibited the phosphorylation of Aurora A kinase, which is associated with important processes during mitosis. The presence of a “comet” DNA fragment and phosphorylation of p53 at Ser 15 clearly indicated that DNA damage occurred with WJ1376-1 and WJ1398-1 treatment. WJ1376-1 and WJ1398-1 ultimately induced apoptosis as evidenced by the upregulation of Bad and activation of caspases-3, -7, and -9. Furthermore, WJ1376-1 and WJ1398-1 also showed a great effect in attenuating tumor growth without affecting the body weight of xenograft nude mice. Taken together, these results suggest that the toxic activities of WJ1376-1 and WJ1398-1 were dissimilar to that of the parental osthole, which can induce cell polyploidy and G₂/M cell cycle arrest in colon adenocarcinoma cells and may provide a potential therapeutic target for colon cancer treatment in the future

Biochemistry tests including GOT, GPT, albumin, BUN, and creatinine.

<p>Biochemistry tests including GOT, GPT, albumin, BUN, and creatinine.</p

Data_Sheet_1.PDF

<p>Growing evidence shows that hydroxamate-based compounds exhibit broad-spectrum pharmacological properties including anti-tumor activity. However, the precise mechanisms underlying hydroxamate derivative-induced cancer cell death remain incomplete understood. In this study, we explored the anti-tumor mechanisms of a novel aliphatic hydroxamate-based compound, WMJ-J-09, in FaDu head and neck squamous cell carcinoma (HNSCC) cells. WMJ-J-09 induced G2/M cell cycle arrest and apoptosis in FaDu cells. These actions were associated with liver kinase B1 (LKB1), AMP-activated protein kinase (AMPK) and p38 mitogen-activated protein kinase (p38MAPK) activation, transcription factor p63 phosphorylation, as well as modulation of p21 and survivin. LKB1-AMPK-p38MAPK signaling blockade reduced WMJ-J-09’s enhancing effects in p63 phosphorylation, p21 elevation and survivin reduction. Moreover, WMJ-J-09 caused an increase in α-tubulin acetylation and interfered with microtubule assembly. Furthermore, WMJ-J-09 suppressed the growth of subcutaneous FaDu xenografts in vivo. Taken together, WMJ-J-09-induced FaDu cell death may involve LKB1-AMPK-p38MAPK-p63-survivin signaling cascade. HDACs inhibition and disruption of microtubule assembly may also contribute to WMJ-J-09’s actions in FaDu cells. This study suggests that WMJ-J-09 may be a potential lead compound and warrant the clinical development in the treatment of HNSCC.</p

Tumor growth and body weight of the orthotopic breast cancer model mice treated with IR (4 Gy) or SAHA (25 mg/kg×9) alone or in combination.

<p>(A) Body weight in Balb/c mice measured once per week. (B) The 4T1-luc cells were injected into mammary fat pads of Balb/c mice, observed for luciferase signals and photographed using IVIS 200. (C) Quantification of the luciferase signals. *, <i>p</i><0.05, versus control. (D) IHC staining of the mouse orthotopic tumor tissues. IHC was used to determine the expression levels of LC3 and γH2AX (×100 objective magnification). The percentage of LC3 and γH2AX-positive cells was determined using HistoQuest software (TissueGnostics).</p

SAHA inhibits experimental metastasis in Balb/c mice.

<p>(A) Body weight in Balb/c mice measured once per week. (B) Lung weights of mice. *, <i>p</i><0.05, versus control. (C) The 4T1-luc cells were observed for the luciferase signals and photographed using IVIS 200. (D) Histological characteristics of mouse lungs stained with H&E. Tumor metastasis is indicated by arrows.</p