OBJECTIVES: To evaluate whether garlicin post-conditioning can attenuate myocardial ischemiareperfusion injury in a catheter-based porcine model of acute myocardial infarction (AMI) by affecting adhesion molecules integrin β1/CD29 and platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31). METHODS: Twenty-two swine were devided into 3 groups: 6 in a sham-operation group, and 8 each in the model and garlicin groups. AMI porcine model was established in the model and garlicin groups. The distal parts of the left anterior descending coronary artery in the animals of the model and garlicin groups were occluded by dilated balloon for 2 h, followed by reperfusion for 3 h. Garlicin (1.88 mg/kg) was injected over a period of 1 h, beginning just before reperfusion, in the garlicin group. Real-time polymerase chain reaction, immunohistochemistry and Western blot were carried out to detect mRNA and protein expressions of CD29 and CD31 3 h after reperfusion. RESULTS: Hematoxylin-eosin staining showed a better myocardial structure in the garlicin group after reperfusion. Compared to the model group, garlicin inhibited both the mRNA and protein expression of CD29 and CD31 in reperfusion area and no-reflflow area (P<0.05 respectively). CONCLUSIONS Garlicin post-conditioning induced cardio-protection against myocardial ischemia-reperfusion injury in this catheter-based porcine model of AMI. The cardio-protective effect of garlicin is possibly owing to suppression of production of CD29 and CD31, by inhibition of the mRNA expression of CD29 and CD31.
Allyl Compounds ; pharmacology ; Animals ; Disease Models, Animal ; Disulfides ; pharmacology ; Integrin beta1 ; analysis ; genetics ; physiology ; Ischemic Postconditioning ; Male ; Myocardial Reperfusion Injury ; prevention & control ; Platelet Endothelial Cell Adhesion Molecule-1 ; analysis ; antagonists & inhibitors ; genetics ; RNA, Messenger ; analysis ; Swine

OBJECTIVETo investigate the effect of diallyl disulfide (DADS) on invasion and metastasis of human breast cancer MCF-7 cells and explore the possible mechanism.

METHODSMCF-7 cells treated with 100, 200, and 400 µmol/L of DADS for 24 h were examined for cell invasion and migration capacities using Transwell assay and wound healing assay, respectively. The protein expression of E-cadherin, vimentin, MMP-9 and p-p38 in the cells were detected with Western blotting. The effect of transforming growth factor-β1 (TGF-β1) as the agonist of p38 activity was tested in antagonizing the effects of DADS.

RESULTSDADS inhibited the invasion and migration of MCF-7 cells in a dose-dependent manner, down-regulated the protein expression of Vimentin and MMP-9 and up-regulated E-cadherin expression in the cells. Treatment with TGF-β1 to up-regulate p38 activity obviously antagonized the inhibitory effect of DADS on the invasion and metastasis of MCF-7 cells.

CONCLUSIONDADS can inhibit the invasion and metastasis of MCF-7 cells in vitro by down-regulating p38 activity.

Allyl Compounds ; pharmacology ; Breast Neoplasms ; pathology ; Cadherins ; metabolism ; Disulfides ; pharmacology ; Gene Expression Regulation, Neoplastic ; Humans ; MAP Kinase Signaling System ; drug effects ; MCF-7 Cells ; drug effects ; Matrix Metalloproteinase 9 ; metabolism ; Mitogen-Activated Protein Kinase 11 ; metabolism ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Transforming Growth Factor beta1 ; pharmacology ; Vimentin ; metabolism

OBJECTIVETo explore the effect of diallyl disulfide (DADS) on hippocampal synapses and learning and memory abilities in a mouse model of A1zheimer's disease (AD).

METHODSMouse models of AD established by agglutinated Aβ1-42 injection in the lateral cerebral ventricle were randomized into 4 groups and treated with DADS at the daily doses of 0, 10, 50 and 100 mg/kg by gavage for 30 consecutive days. The learning and memory abilities of the mice were assessed with Morris water maze test; the structures of the dendritic spines and synapses in CA1 region of the hippocampus were observed under transmission electron microscope with silver staining; PSD95 and SYP protein and mRNA expressions in the hippocampus were detected with Western blotting and RT-PCR.

RESULTSCompared with the AD model mice, the mice treated with 50 and 100 mg/kg DADS showed enhanced learning and memory abilities in Morris water maze test. The dendritic spines and synapses in CA1 region of the hippocampus increased obviously and hippocampal expressions of PSD95 and SYP were enhanced in mice treated with 50 and 100 mg/kg DADS.

CONCLUSIONDADS at the daily doses of 50 and 100 mg/kg can improve the learning and memory abilities and increase the number of dendritic spines and synapses in the hippocampus in mouse models of AD.

Allyl Compounds ; pharmacology ; Alzheimer Disease ; drug therapy ; Animals ; CA1 Region, Hippocampal ; drug effects ; Disease Models, Animal ; Disulfides ; pharmacology ; Learning ; Male ; Memory ; Mice ; Synapses ; drug effects

OBJECTIVETo study the role of the PI3K/AKT signaling pathway in the diallyl disulfide (DADS)-induced apoptosis of K562 cells.

METHODSK562 cells in the logarithmic growth phase were treated with 10, 20, 40, or 80 mg/L DADS for 48 hours, then fixed and stained with acridine orange/ethidium bromide (AO/EB), and examined for cellular morphological changes under an inverted microscope. Annexin V-FITC/PI staining was used for determining the apoptotic rates, and Western blot for measuring the expression of AKT, p-AKT, and Caspase-3. Two control groups, blank and solvent, were used as references.

RESULTSK562 cells treated with DADS for 48 hours exhibited the characteristic morphological features of apoptosis including cell shrinkage, irregular cell shape, and membrane blebbing. AO/EB staining results demonstrated that the number of apoptotic cells with cell shrinkage, pyknotic or bead-like nuclei, chromatin condensation, and orange staining increased with the increasing DADS concentration, and 40 mg/L DADS had the most significant effect. The apoptotic rates of cells treated with 10, 20, 40, and 80 mg/L DADS were all significantly higher than those in the control groups (P<0.05). There were no significant differences in AKT protein expression between the K562 cells treated with different concentrations of DADS; the p-AKT protein expression decreased with the increasing DADS concentration, while the Caspases-3 protein expression increased with the increasing DADS concentration (P<0.05).

CONCLUSIONSDADS induces the apoptosis of K562 cells, probably through inhibiting the protein expression in the PI3K/AKT signaling pathway.

Allyl Compounds ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Disulfides ; pharmacology ; Dose-Response Relationship, Drug ; Humans ; K562 Cells ; Phosphatidylinositol 3-Kinases ; physiology ; Proto-Oncogene Proteins c-akt ; physiology ; Signal Transduction ; physiology

Hypoxia is a general characteristic of most solid malignancies and intimately related to cancer progression. Homeostatic response to hypoxia is primarily mediated by hypoxia inducible factor-1alpha (HIF-1alpha) that elicits transcriptional activity through recruitment P300 coactivator. Targeting the interaction of HIF- alpha and P300 would thus constitute a novel approach for cancer treatment by suppressing tumor angiogenesis and metastasis. Here, a screening assay was developed for inhibitors targeting the interaction between HIF-1alpha and P300. The nucleotide sequence of human HIF-1alpha and P300 were cloned into pBIND and pACT vectors, named pBIND-HIF1alpha and pACT-P300. The interaction of HIF-1alpha and P300 was identified in HEK293 cell using mammalian two-hybrid system. And compound chetomin decreased their interaction in this mammalian two-hybrid system. We further verified HIF-1 inhibition effect of chetomin in U251-HRE cells. Therefore, we established a screening assay combined HIF-1alpha and P300 mammalian two-hybrid system and U251-HRE reporter assay for HIF-1 selective inhibitors.
Cell Hypoxia ; Disulfides ; pharmacology ; Drug Screening Assays, Antitumor ; E1A-Associated p300 Protein ; antagonists & inhibitors ; HEK293 Cells ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; antagonists & inhibitors ; Indole Alkaloids ; pharmacology ; Two-Hybrid System Techniques

Antineoplastic Agents ; pharmacology ; Autophagy ; drug effects ; Disulfides ; pharmacology ; Gene Expression Regulation, Neoplastic ; Glycogen Synthase Kinase 3 ; genetics ; metabolism ; HCT116 Cells ; Humans ; Piperazines ; pharmacology ; Proto-Oncogene Proteins ; genetics ; metabolism ; Proto-Oncogene Proteins p21(ras) ; Signal Transduction ; ras Proteins ; genetics ; metabolism

OBJECTIVETo evaluate whether garlicin can prevent reperfusion no-reflow in a catheter-based porcine model of acute myocardial infarction (AMI).

METHODSTwenty-two male Chinese mini swines were randomized into 3 groups: sham-operation group (n=6), control group (n=8), and garlicin group (n=8). The distal part of left anterior descending coronary artery (LAD) in swines of the latter two groups was completely occluded by dilated balloon for 2 h and a successful AMI model was confirmed by coronary angiography (CAG) and electrocardiograph (ECG), which was then reperfused for 3 h. In the sham-operation group, balloon was placed in LAD without dilatation. Garlicin at a dosage of 1.88 mg/kg was injected 10 min before LAD occlusion until reperfusion for 1 h in the garlicin group. To assess serial cardiac function, hemodynamic data were examined by catheter method before AMI, 2 h after occlusion and 1, 2, and 3 h after reperfusion. Myocardial contrast echocardiography (MCE) and double staining with Evans blue and thioflavin-S were performed to evaluate myocardial no-reflow area (NRA) and risk area (RA).

RESULTSLeft ventricular systolic pressure and left ventricular end-diastolic pressure significantly improved in the garlicin group after reperfusion compared with the control group P<0.05) and 2 h after AMI (P<0.05). MCE showed garlicin decreased reperfusion NRA after AMI compared with the control group (P <0.05). In double staining, NRA/RA in the garlicin group was 18.78%, significantly lower than that of the control group (49.84%, P<0.01).

CONCLUSIONSGarlicin has a preventive effect on the porcine model of myocardial infarction reperfusion no-reflow by improving hemodynamics and decreasing NRA.

Allyl Compounds ; pharmacology ; therapeutic use ; Animals ; Cardiotonic Agents ; pharmacology ; therapeutic use ; Contrast Media ; Disease Models, Animal ; Disulfides ; pharmacology ; therapeutic use ; Hemodynamics ; drug effects ; Male ; Myocardial Infarction ; complications ; diagnostic imaging ; drug therapy ; pathology ; Myocardial Reperfusion ; No-Reflow Phenomenon ; complications ; diagnostic imaging ; drug therapy ; pathology ; Swine ; Swine, Miniature ; Thiazoles ; metabolism ; Ultrasonography

OBJECTIVETo observe whether garlicin could ameliorate pressure overload induced myocardial fibrosis in rats through partial inhibiting transforming growth factor beta1 (TGF-beta1) mediated Smads signal.

METHODSForty male SD rats were randomly divided into 4 groups, i. e., the sham-operation group, the model group, the garlicin group, and the Tetramethylpyrazine (TMP) group, 10 in each group. The pressure overload induced myocardial fibrosis rat model was prepared using coarctation of aorta. Three days after modeling 5.0 mg/kg garlicin injection was administered to rats in the garlicin group, 20 mg/kg TMP injection to rats in the TMP group by peritoneal injection, while normal saline was peritoneally injected to rats in the sham-operation group and the model group. Four weeks after medication, the changes of myocardial collagen were observed by picrosirius red staining. The myocardial collagen volume fraction (CVF) and perivascular collagen areas (PVCA) were calculated. The serum transforming growth factor beta1 (TGF-beta1) expression was detected using ELISA. The TGF-beta1 protein expression in the myocardial tissue was observed using immunohistochemical assay. The changes of myocardial Smad2 and Smad7 mRNA expressions were detected using Real-time RT-PCR. The effects of garlicin on TGF-beta1 mediated Smad Signaling through luciferase assay were further verified using Mv1 Lu-(CAGA) 12-Luc cell line response to TGF-beta1.

RESULTSCompared with the sham-operation group, the myocardial levels of CVF and PVCA, the serum TGF-beta1 level, and the TGF-beta1 protein expression in the myocardial tissue obviously increased in the model group (P < 0.05, P < 0.01). Compared with the model group, the PVCA level, the serum TGF-beta1 level, and the TGF-beta1 protein expression in the myocardial tissue of the garlicin group and the TMP group obviously decreased (P < 0.05, P 0O 01). The Smad2 mRNA expression was up-regulated while Smad7 mRNA expression down-regulated in the model group. The Smad2 mRNA expression was obviously down-regulated in the garlicin group and the TMP group (P < 0.05). The Smad7 mRNA expression was obviously up-regulated in the TMP group (P > 0.05). One to 2 microg/mL garlicin could obviously inhibit the luciferase activities of corresponding TGF-beta1, under the stimulation of 2 ng/mL TGF-beta1 (P < 0.05).

CONCLUSIONGarlicin ameliorated pressure overload induced myocardial fibrosis in rats through partial inhibiting TGF-beta-Smads signal pathway.

Allyl Compounds ; pharmacology ; Animals ; Cardiomyopathies ; etiology ; metabolism ; pathology ; Disulfides ; pharmacology ; Fibrosis ; Male ; Myocardium ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Smad Proteins ; metabolism ; Transforming Growth Factor beta1 ; metabolism

Recent years, the incidence and mortality of prostate cancer have increased dramatically in China. At earlier stages, most diagnosed prostate cancers are responsive to androgen depletion treatment, yet, nearly all patients will eventually progress to metastatic androgen-independent prostate cancer (AIPC), which still has no effective therapeutic method or drug to deal with. 11'-Deoxyverticillin A (C42) belongs to the family of epipolythiodioxopiperazines (ETPs), an interesting class of fungal toxins that inhibit farnesyl transferase. Compounds holding such a property have been explored as putative anticancer agents. In this study, using PC3M cells, an AIPC cell line, we investigated the effect of the compound on apoptosis and explored the underlying mechanism. It revealed that C42 markedly enhanced the activity of caspase-3/7 and increased the accumulation of the cleaved PARP, all of which are the markers of apoptosis. It also revealed that C42 either decreased cell viability or inhibited the growth of PC3M cells. Moreover, we observed that the loss of cell viability and cell growth inhibition induced by C42 were both time- and dosage dependent. Taken together, we indicated that C42 can induce caspase-dependent apoptosis in AIPC cells, and the results presented here will broaden our knowledge about the molecular mechanisms by which C42 exerts its anticancer activity, and future work in this direction may provide valuable information in the development of these compounds into effective cancer therapeutic strategies against androgen-independent prostate cancer.
Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 7 ; metabolism ; Cell Line, Tumor ; Disulfides ; pharmacology ; Farnesyltranstransferase ; antagonists & inhibitors ; Humans ; Male ; Mycotoxins ; pharmacology ; Piperazines ; pharmacology ; Prostatic Neoplasms ; pathology

OBJECTIVETo study the effects of diallyl disulfide (DADS) on apoptosis of human leukemia K562 cells and possible mechanisms.

METHODSThe morphologic changes of leukemia K562 cells after DADS treatment were observed by Hoechst 33258 staining. Cell apoptosis rates after different concentrations and different durations of DADS treatment were determined by flow cytometry. Fas, FasL and caspase-8 mRNA expression was estimated by reverse transcription-polymerase chain reaction (RT-PCR) 48 hrs after DADS treatment.

RESULTSThe characteristics of apoptosis in K562 cells induced by DADS were observed. After 24 hrs of DADS treatment, the apoptosis rate of K562 cells increased from (11.60 ± 0.83)% at the concentration of 10 mg/L to (37.94 ± 0.87)% at the concentration of 40 mg/L. The apoptosis rate of K562 cells increased after 40 mg/L DADS with the increasing time from (37.94 ± 0.87)% (24 hrs) to (47.02 ± 0.66)% (72 hrs). Expression of Fas and caspase-8 mRNA increased, while FasL mRNA expression decreased significantly 48 hrs after DADS treatment compared with the control group (P<0.05).

CONCLUSIONSDADS can induce apoptosis of human leukemia K562 cells in a time- and concentration-dependent manner, possibly through increasing Fas and caspase-8 expression and decreasing FasL expression.

Allyl Compounds ; pharmacology ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Bisbenzimidazole ; Caspase 8 ; genetics ; Disulfides ; pharmacology ; Fas Ligand Protein ; genetics ; Flow Cytometry ; Humans ; K562 Cells ; RNA, Messenger ; analysis ; fas Receptor ; genetics