OBJECTIVETo explore the protective effect of allicin on nicotine-induced oxidative damage to human periodontal ligament cells (HPDLCs).

METHODS(1) Establish nicotine-induced oxidative damage model on HPDLCs. Use water-soluble tetrazolium (WST) colorimetric method to find out the nicotine concentration (X) that could inhibit HPDLCs' growth for the following experiments. (2) HPDLCs of the fifth passage were divided into 5 groups: The control group, the nicotine group and the nicotine+allicin groups(the concentration of allicin was 15, 30, and 60 microg x mL(-1) respectively). Different kinds of culture media were added. Similarly, use WST colorimetric method to choose the allicin concentration (Y) that could significantly improve the survival rate of HPDLCs. (3) HPDLCs were divided into 3 groups: The control group, the nicotine group, the nicotine+allicin group and different media were added. The glutathion (GSH) concentrations in HPDLCs were determined in 1, 4, 8, 12 and 24h respectively.

RESULTS0.8 mg x mL(-1) nicotine could inhibit the HPDLCs survival rate significantly (77% of the control, P < 0.05). But 60 microg x mL(-1) allicin could prevent the inhibition effects evidently, improving the survival rate to 112% of that of the nicotine group (P < 0.05) and reaching the survival rate level of control group (P > 0.05). The GSH concentrations of nicotine+allicin group were higher than that of the nicotine group always (P < 0.05) and by 82% at 8 h after culture, but had no difference with that of the control group (P > 0.05).

CONCLUSION60 microg x mL(-1) allicin can protect the HPDLCs against oxidative damage induced by nicotine.

Cell Proliferation ; Cells, Cultured ; Culture Media ; Humans ; Nicotine ; Periodontal Ligament ; Sulfinic Acids

To investigate the effects of allicin on rats by NMR-based metabonomic method, the changes of endogenous metabolites in normal rat urine and the influences on metabolism were analyzed with bio-nuclear magnetic resonance (NMR) method and partial least-squares discriminant analysis (PLS-DA) after intraperitoneal administration of allicin solution. The identified biochemical effects associated with allicin dosing included elevated then gradually recovered urinary levels of Kreb's cycle intermediates, such as citrate, alpha-ketoglutarate and succinate and increased concentrations of ketones. Meanwhile, decreased urinary concentrations of glucose, lactate, alanine, hippurate and trimethylamine oxide were observed. The PLS-DA revealed that the metabonomic profiles of allicin treated groups were obviously different from those of the control group. Allicin may change metabolism significantly in normal rats. The study of the pharmacologic mechanism of allicin by metabonomic method is practicable and it could be explored further.
Animals ; Magnetic Resonance Spectroscopy ; Male ; Metabolomics ; Rats ; Rats, Sprague-Dawley ; Sulfinic Acids ; metabolism ; urine

Adenosine Triphosphatases ; metabolism ; Animals ; Antioxidants ; metabolism ; Muscle, Skeletal ; drug effects ; enzymology ; Rats ; Sulfinic Acids ; pharmacology

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Sulfinic Acids ; pharmacology

OBJECTIVE: To observe the anticancer mechanism of allicin by observing the inhibitory effect of allicin on human salivary gland carcinoma cell line MEC-1. METHOD: Cell proliferation were measured by MTT assay at different doses and different hours. In the meantime, cell cycle was detected via flow cytometry after different dose incubation with different hours. RESULT: MTT results showed that the inhibitory rates of MEC-1 proliferations were increased in a concentration-and time-dependent manner. Flow cytometry analysis showed percent-age of MEC-1 cells decreased in G0/G1 phase and increased in G2/M. But there was no evident change in S phase. The cells were mainly blocked in M phase, and the inhibitory effect of the allicin on MEC-1 cells increased with the increasing of concentration and time. CONCLUSION Allicin can inhibit the growth of MEC-1 cells in vitro dramatically.
Cell Cycle ; drug effects ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; Humans ; Sulfinic Acids ; pharmacology

OBJECTIVEThe objective of this paper was to study the change of P38MAPK and Fas in the apoptosis of THP-1 cells induced by allicin.

METHODThe proliferation inhibition rates of THP-1 cells after various treatments were examined by MTT assay. Apoptosis rate was determined with Annexin V- FITC/PI double staining by flow cytometry. The expression and distribution change of the phosphorylation p38MAPK (P-p38MAPK) were detected by immunohistochemical staining. The changes of P-p38 MAPK and Fas proteins were detected by Western blot.

RESULTThe proliferations of leukemia cell line THP-1 are inhibited by allicin. MTT assay showed that allicin can inhibit the proliferation of the THP-1 cell, and the inhibition was dependent on both dose and time. The IC50 of 72 hours was 12.8 mg x L(-1). Apoptosis rate detected by Annexin V-FITC/PI was proportional to the concentration of the allicin. After the immunohistochemical staining test, the P-p38MAPK was located in the cell nucleus and plasma, showing deep brown, when adding allicin to THP-1 cell. Western blot test showed that the P-p38MAPK proteins expression was proportional to the concentration of Allicin and was also dose dependent. The levels of P-p38MAPK in negative control group, 1/2 IC50 of 72 hours group and IC50 of 72 hours group were 0.259 8 +/- 0.013 2, 0.61 2 +/- 0.008 3 and 0.505 6 +/- 0.005 5 respectively, and the levels of Fas proteins were 0.287 4 +/- 0.008 9, 0.426 8 +/- 0.007 9 and 0.597 1 +/- 0.010 9 respectively. The difference was statistically significant when compared with the negative control group (P < 0.01).

CONCLUSIONAllicin can significantly induce THP-1 cells apoptosis, and its mechanism may be related to the activation of P38MAPK/Fas.

Apoptosis ; drug effects ; Cell Line, Tumor ; Humans ; Neoplasms ; drug therapy ; metabolism ; physiopathology ; Phosphorylation ; Signal Transduction ; drug effects ; Sulfinic Acids ; pharmacology ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism

Animals ; Apoptosis ; drug effects ; Brain Ischemia ; pathology ; Hippocampus ; cytology ; drug effects ; pathology ; Neurons ; drug effects ; pathology ; Rats ; Rats, Wistar ; Reperfusion Injury ; pathology ; Sulfinic Acids ; pharmacology

To analyze the chemical components and decomposition products in allicin extract of garlic, the chemical components screening and identification were made with HPLC-MS/MS method by full scan TIC MS, HPLC retention time, product MS spectra and chemical reference standards. The stability of the extract in water and alcoholic solutions was also investigated. There were five major components in allicin extract which were all identified as thiosulfinates. The extract was stable for at least 3 months when stored at -20 degrees C as water solution, but obvious decomposition was observed with the increase of alcoholic concentration. The decomposition products were also identified by HPLC-MS/MS.
Chromatography, High Pressure Liquid ; Drug Stability ; Garlic ; chemistry ; Plants, Medicinal ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; Sulfinic Acids ; isolation & purification ; metabolism ; Tandem Mass Spectrometry ; Thiosulfates ; analysis

OBJECTIVETo investigate the effect of allicin in inhibiting insulin-induced vascular smooth muscle cell (VSMC) proliferation and migration.

METHODThe tissue explant method was adopted to culture rat's aVSMCs, and the immunofluorescence method was used to identify α-SMA. Cells from the third to fifth generations were selected in the experiment The insulin-induced VSMC model was established. The experiment was carried out in five groups: the control group, the insulin group, the allicin group, the ERK inhibitors PD98059 group (20 μmol · L(-1)) and the PD98059 + allicin group. VSMCs' proliferation was determined by CCK8 colorimetric method, while its migration was detected by cell counting; The western blotting was used to detect total ERK, Phospho-ERK, PCNA protein's expression.

RESULTPrimary cultured VSMCs grew well in the spindle shape under the lightmicroscope, with peak and valley. α-SMA immunofluorescence results showed that the cultured cells had typical VSMCs' features. Insulin could stimulate VSMCs' proliferation and migration, with the best effect at the concentration of 100 nmol · L(-1). The pretreatment with allicin could significantly inhibit VSMCs' proliferation and migration induced by insulin in a dose-dependent manner. The pretreatment with PD98059 and allicin + PD98059 could inhibit VSMCs' proliferation and migration induced by insulin remarkably as well. Insulin could significantly accelerate VSMCs' expression of such proteins as p-ERK, PCNA. Contrarily, allicin could notably inhibit VSMCs' expression of such proteins as p-ERK, PCNA in a dose-dependent manner.

CONCLUSIONAllicin could significantly inhibit VSMCs' proliferation and migration induced by insulin, which may be related to the inhibition of the activation of ERK signal path.

Animals ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Insulin ; metabolism ; Male ; Muscle, Smooth, Vascular ; cytology ; drug effects ; metabolism ; Myocytes, Smooth Muscle ; cytology ; drug effects ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Sulfinic Acids ; pharmacology

OBJECTIVETo study the effect of allicin on human colon cancer cell line LoVo and the combined effect of allicin and CPT-11 on this cancer cell line.

METHODThe LoVo cells were cultured in vitro and treated with allicin in different concentrations. MTT assay was used to test dynamically the cell growth inhibiting effect. Apoptosis induction (Annexin-V-FITC/PI) and modulation of DNA cell cycle were measured by flow cytometry. The change of cytotoxicity of CPT-11 after combination of allicin at the concentration of 4.0, 8.0 mg x L(-1) were investigated.

RESULTAllicin had inhibitive effect on growth of LoVo cells in a dose and time dependent manner, with IC50 value of 32.23, 10.74, 6.58 mg x L(-1) at 24 h, 48 h and 72 h, respectively. The apoptosis rate of LoVo cells increased progressively as the cells were treated with increasing concentration of allicin in 24 h, while the apoptosis rate achieved peak value when the cells were treated with allicin at the concentration of 8 mg x L(-1) in 48 h. The result indicated the low concentrations of allicin (< 4 mg x L(-1)) lead to G2/M cell cycle arrest, and higer concentrations ( > 4 mg x L(-1)) exert G1 + G2/M cell cycle arrest in 24 h. Compared with single use of CPT-11, the combined use of CPT-11 and allicin (4.0, 8.0 mg x L(-1), respectively) showed increasing cytotoxicity on the LoVo cells, with IC50 of 24 h decreasing from 47.5 to 7.4 and 7.2 mg x L(-1), respectively.

CONCLUSIONAllicin has significant anti-proliferation effect on human colon cancer cell line LoVo by induction of apoptosis and arrestment of cell cycle and can enhance the cytotoxicity of CPT-11 on the colon cancer LoVo cell.

Antineoplastic Agents, Phytogenic ; toxicity ; Apoptosis ; drug effects ; Camptothecin ; analogs & derivatives ; toxicity ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Colonic Neoplasms ; drug therapy ; physiopathology ; Humans ; Models, Biological ; Sulfinic Acids ; pharmacology