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

BACKGROUND: Smad7 is a major repressible protein in transforming growth factor β(TGF-β1) signal transduction pathway,which possess antifibrotic effects.OBJECTIVE: To construct rat Smad7 eukaryotic vector and to observe the mRNA expression level of TGF-β1, collagen Ⅰ and collagen Ⅲ in rat hepatic stellate cells (HSC)-T6 cell.DESIGN, TIME AND SETTING: The gene recombination and cytology observation experiment was performed at the First Affiliated Hospital of Shihezi University School of Medicine.MATERIALS: pcDNA3.1 (+) plasmid was reserved in the laboratory. E coil DH5α was presented by Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Shihezi University School of Medicine. The HSC T6 cell was provided by Cancer Institute and Hospital, Chinese Academy of Medical Sciences.METHODS: Rat Smad7 cDNA was cloned into eukaryotic plasmid pcDNA3.1(+) to construct Smad7/pcDNA3.1(+) plasmid and transfected it into HSC-T6 ceils by Lipofectmine 2000. The experiment was divided into normal control, empty vector and Smad7 transfected groups, and the positive cells were selected by G418.MAIN OUTCOME MEASURES: The levels of Smad7, TGF-β1, collagen Ⅰ and Ⅲ mRNA was detected by reverse transcriptase polymerase chain reaction, respectively.RESULTS: Smad7 eukaryotic vector was successfully constructed and confirmed by endonuilease digestion and sequencing. Compared to the control and empty vector groups, Smad7 mRNA expression was significantly higher in Smad7 transfected group (P < 0.01 ); and TGF-β1 and collagen Ⅰ mRNA expression was notably reduced (P < 0.01). There was no statistically significant difference of the change of collagen Ⅲ mRNA expression among the three groups (P>0.05). The difference of Smad7, TGF-β1,collagen Ⅰ and Ⅲ mRNA expression had no statistically significant between control and empty vector groups (P_(all) > 0.05)CONCLUSION: Smad7 eukaryotic expression vector is successfully constructed. The Smad7 gene can effectively expressed in transfected HSC-T6 cell, and decrease mRNA expressions of TGF-β1 and collagen Ⅰ.

Child ; Congresses as Topic ; Humans ; Pediatrics

Child ; Congresses as Topic ; Humans ; Pediatrics

Humans ; Microcirculation ; Sepsis ; physiopathology

Background Recently,there were many studies on corneal innervations during mammalian development.However,there were fewer studies on discussing corneal innervations before and after mouse eye openings.Objective The present study was to investigate the change in the regulation of corneal nerve fiber length and density before and after mouse eye openings to offer a basis for clinical research in human.Methods Thirty SPF C57BL/6 mice were divided into postnatal 1 day(P1 d),P7 d,P13 d(1 day before eye opening),P14 d(eye halfopened),P17 d(1 day after eye opening)and P23 d(7 day after eye opening)groups,with 5 mice and 10 eyes for each group.Entire corneal stretches were prepared and immunostaining with an anti-neuron-specific β-Ⅲ tubulin antibody was performed to label the corneal nerve fibers.Confocal microscopic pictures from the corneal dorsal-nasal region (DN),dorsal-temporal(DT),ventral-nasal region(VN)and ventral-temporal(VT)were taken using Delta Vision Core.From these pictures,the mouse corneal area,total length and density of nerve fibers in the 4 regions were calculated.The use of the animals complied with Statement of ARVO.Results Corneal areas of P1 d,P7d,P13 d,P14 d,P17 d and P23 d mice were(0.404±0.007),(1.362±0.154),(1.573±0.080),(1.603±0.046),(1.847±0.052),(2.445±0.798)mm2,respectively ; the total lengths of nerve fibers were(3.718±1.044),(19.065±3.350),(23.687±0.907),(27.309±2.477),(31.989±3.976),(41.214±1.573)mm,respectively ; the densities of nerve fibers were(9.592±1.138),(14.506±1.908),(15.088±1.241),(16.772±1.897),(16.821±2.102),(17.660±1.216)mm/mm2,respectively,all showing significant increases with age(F =22.906,P =0.000 ; F =0.424,P =0.000 ; F =2.375,P=0.000).A positive correlation of the increasing corneal areas and increasing lengths of nerve fibers was found(r=0.983,P＜0.01).Nerve fiber densities in the four corneal regions significantly increased with age(DN region:F =0.159,P =0.000 ; DT region:F =2.1 72,P =0.001 ; VN region:F =1.998,P =0.000 ; VT region:F=2.352,P=0.000).From P13 d to P14 d,the corneal nerve fiber densities in the DN region decreased by 6.0％ without significant difference(t =0.589,P =0.572); and the corneal nerve fiber densities in the DT region,VN region and VT region decreased by 4.6％,5.5％ and 0.1％,respectively,without significant difference from P14 d to P17 d(t=0.549,P=0.596;t=0.701,P=0.501 ;t=-0.100,P=0.919).Conclusions The development of nerve fibers in the whole cornea or the four corneal regions is influenced by eye opening in mouse to various extents.From P13 d to P14 d,the corneal nerve fiber densities in the DN region decreased by 6.0％ without significant difference.From P14 d to P17 d,the corneal nerve fiber densities in the DT region,VN region and VT region decrease by 4.6％,5.5％ and 0.1％,respectively,without significant difference.Afterwards,the growth of nerve fibers increased in pace and the growth rate is recovered.

0.05). But the LDH 5 and LDH 5/LDH 1 of difference doses of Shenmai Injection group was lower than that of the control group obviously(P

To study the compatible mechanisms and compatible proportion of Shaoyao Gancao decoction, the intestinal absorption of main ingredients in Shaoyao Gancao decoction SG11 (Baishao-Zhigancao 1: 1) , SG31 (Baishao-Zhigancao 3: 1), Baishao water decoction S and Zhigancao (G) were investigated and compared using in vitro everted intestinal sac model and in situ single pass intestinal perfusion (SPIP) model. The concentration of paeoniflorin (PF), liquiritin (LQ) and mono-ammonium glycyrrhizinate (GL) in test samples and samples of intestinal sac and intestinal perfusion was determined by HPLC. The intestinal absorptive amount and absorption parameters were calculated. Results showed that in the everted intestinal sac model, three ingredients could be absorbed by duodenum, jejunum and ileum, and the absorption in the jejunum was best for all 3 ingredients. The absorption rate of three ingredients in SG11 was significantly higher than that in single decoction (P < 0.05), but had no significant difference compared with SG31. In SPIP model, the absorption rate constant K(a), the apparent absorption coefficient P(app) and the absorption rate of three ingredients in SG11 were significantly higher than those in single decoction. Parameters of PF and GL in SG11 were significantly higher than those in SG31, but had no differences of LQ. It proved that the compatibility of Baishao and Zhigancao could improve the intestinal absorption of PF, LQ and GL. The absorption of each ingredient in SG11 was better than that in SG31.
Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; pharmacokinetics ; Intestinal Absorption ; drug effects ; Intestines ; blood supply ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the intestinal absorption mechanism of tilianin in rats.

METHODThe single-pass perfusion model was established in rats. The concentrations of tilianin with in situ intestinal perfusion were determined by HPLC. The impact factors, such as verapamil, reserpine, phloridzin and rifampicin, on Ka and Papp of tilianin in rat jejunum were investigated.

RESULTCompared with the control group, Ka and Papp in rat jejunum were significantly higher after being added with verapamil and reserpine (P < 0.05). Papp of tilianin in rat jejunum was significantly lower after being added with phloridzin (P < 0.05). Compared with the control group, both Ka and Papp of tilianin in rat jejunum were not significantly higher after being added with rifampicin.

CONCLUSIONTilianin is the substrate of P-gp, BCRP and SGLT1. The effluent effect of P-gp and BCRP is the main mechanism of tilianin in intestinal absorption, indicating that tilianin can realize intestinal absorption and transport by relying on SGLT1. Tilianin is not the substrate of bile salt transporter protein.

Animals ; Caco-2 Cells ; Drugs, Chinese Herbal ; pharmacokinetics ; Female ; Flavonoids ; pharmacokinetics ; Glycosides ; pharmacokinetics ; Humans ; Intestinal Absorption ; Lamiaceae ; chemistry ; Male ; Models, Biological ; Perfusion ; Rats ; Rats, Wistar