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1.
China Journal of Chinese Materia Medica ; (24): 4857-4863, 2019.
Article Dans Chinois | WPRIM | ID: wpr-1008174

Résumé

To prepare Helix aspersa muller-paeonol nanogel( PAE-HAM-Gels) with anti-proliferative scar effect,evaluate its skin penetration,retention and irritation,and to investigate its prevention and treatment effect for hypertrophic scar in rabbit ears. The dermal retention,transdermal rate and cumulative permeability of paeonol were investigated in vitro by using the modified Franz diffusion cell and the abdominal skin of suckling pigs,SD rats and KM mice,respectively,and the in vitro permeation curves were drawn. The normal skin of the back of New Zealand rabbits was continuously treated with PAE-HAM-Gels for 7 days,and the physiological state of the skin was observed under light microscope after HE staining by using homologous left and right contrast method. The hypertrophic scar model in rabbit ears was established,and the New Zealand rabbits were randomly divided into blank group,model group,positive drug group,PAE-Gels group and PAE-HAM-Gels group. After 28 days of administration,the scar hyperplasia rate and scar elevation index( SEI) of each group were calculated; the scar tissues were taken and stained with Masson for observation of collagen fibers and muscle fibers hyperplasia under light microscope,and the expression level of TGF-β1 in each group was detected. The Qnof PAE-HAM-Gels in aqueous solution was in line with the Higuchi equation,and its transdermal rate,cumulative permeation and dermal retention in different animal skins were all higher than those of PAE-Gels. The skin of the drug-administered group was intact,without erythema,edema or other phenomena; under light microscope,the subcutaneous tissue and the epidermal cells were neatly arranged with uniform thickness,which showed no difference from the blank group. The scar hyperplasia rate of the PAE-HAM-Gels group was 62. 50%; SEI was 2. 17±0. 33 and TGF-β1 was( 815. 4±34. 69) ng·L~(-1),significantly different from those in model group( P<0. 01). Masson staining showed that as compared with the model group,the number of collagen fibers and muscle fibers was small and the arrangement was loose and tidy in the PAE-HAM-Gels group,with regular arrangement of chondrocytes and a small number of inflammatory cells and microvessels.PAE-HAM-Gels have good transdermal properties and dermal retention without skin irritation,offering a promising therapeutic strategy for transdermal delivery during the prevention and treatment of hypertrophic scar in rabbit ears.


Sujets)
Animaux , Souris , Lapins , Rats , Acétophénones/composition chimique , Cicatrice hypertrophique , Oreille , Nanogels/composition chimique , Rat Sprague-Dawley , Suidae
2.
China Journal of Chinese Materia Medica ; (24): 357-363, 2019.
Article Dans Chinois | WPRIM | ID: wpr-774596

Résumé

Hypertrophic scar( HS) is a very common skin fibrosis disorder after human skin injury and wound healing. The objective of this study was to investigate the efficacy of cell penetrating peptide TAT-modified liposomes loaded with salvianolic acid B( SAB-TAT-LIP) on proliferation,migration and cell cycle of human skin fibroblasts( HSF),and preliminarily evaluate its effect on prevention and treatment of HS. HSF were cultured in vitro,and MTT assay was used to detect the inhibitory effect of SAB-TAT-LIP on cell proliferation. Cell migration was assessed by Transwell chamber method and scratch method; and cell cycle change was detected by flow cytometry. In vitro cell studies showed that blank liposome basically had no toxic effect on HSF. Different concentrations of SABTAT-LIP inhibited proliferation on HSF in varying degrees after intervention for different periods in a dose and time dependent manner;meanwhile,SAB-TAT-LIP significantly inhibited the migration and invasion of HSF. At the same time,SAB-TAT-LIP could block the cell cycle at G0/G1 phase after intervention for 48 h,P<0.01 as compared with the blank control group. Conclusively,our experimental data quantitatively demonstrate that SAB-TAT-LIP has significant inhibitory effect on cells proliferation,invasion and migration,with blocking effect on G0/G1 phase. This may offer a promising therapeutic strategy for transdermal delivery in prevention and treatment of HS.


Sujets)
Humains , Benzofuranes , Pharmacologie , Cycle cellulaire , Mouvement cellulaire , Prolifération cellulaire , Peptides de pénétration cellulaire , Cellules cultivées , Vecteurs de médicaments , Fibroblastes , Biologie cellulaire , Liposomes , Peau , Biologie cellulaire
3.
Chinese Traditional and Herbal Drugs ; (24): 59-68, 2019.
Article Dans Chinois | WPRIM | ID: wpr-851439

Résumé

Objective To prepare the liposomes of salvianolic acid B modified with cell penetrating peptide TAT (SAB-TAT-LIP), of which has effects on preventing and treating hypertrophic scars (HS), and establish the method of quality evaluation, as well as preliminarily investigate the effect on the proliferation and migration of human skin fibroblasts (HSF). Methods Liposomes were prepared by pH gradient reverse-phase evaporation method, and the entrapment efficiency was measured by ultrafiltration. Box-Behnken design was performed to optimize the formulation of liposomes by using encapsulation rate as evaluating index. The physicochemical properties of liposomes including morphology, entrapment efficiency, particle size, zeta potential, in vitro release and transdermal absorption, and stability were studied. In addition, the effect of liposomes on proliferation of HSF was examined by MTT assay, and the effect of liposomes on migration of HSF was investigated by scratching method and Transwell assay. Results Based on the optimal formulation of SAB-TAT-LIP, the entrapment efficiency of salvianolic acid B was (86.70 ± 0.85)%, the average particle size was (219.90 ± 5.09) nm, and the zeta potential was (-9.25 ± 0.92). The in vitro 24 h cumulative release was 62.49% of the total drug with no burst effect. The in vitro 32 h cumulated skin penetration rate was 17.21%, the permeance rate was (28.33 ± 4.9) μg/(cm2∙h), and the retention volume of dermis was (44.39 ± 6.87) μg/cm2. The stability was good when placed at 4 ℃ for 10 d. The in vitro cell studies showed that SAB-TAT-LIP can significantly inhibit the proliferation, migration and invasion of human skin fibroblasts, compared with the control group (P < 0.01). Conclusion The optimized SAB-TAT-LIP have higher encapsulation efficiency, smaller particle size, good sustained release effect, and good dermal retention effect which all satisfy the in vitro release and transdermal regulation of local transdermal preparation, and it can significantly inhibit the proliferation, migration and invasion of human skin fibroblasts in vitro.

4.
Asian Journal of Andrology ; (6): 749-757, 2008.
Article Dans Anglais | WPRIM | ID: wpr-359914

Résumé

<p><b>AIM</b>To investigate whether adriamycin induces DNA damage and the formation of gammaH2AX (the phosphorylated form of histone H2AX) foci in mature spermatozoa.</p><p><b>METHODS</b>Human spermatozoa were treated with adriamycin at different concentrations. gammaH2AX was analyzed by immunofluorescent staining and flow cytometry and double-strand breaks (DSB) were detected by the comet assay.</p><p><b>RESULTS</b>The neutral comet assay revealed that the treatment with adriamycin at 2 microg/mL for different times (0.5, 2, 8 and 24 h), or for 8 h at different concentrations (0.4, 2 and 10 microg/mL), induced significant DSB in spermatozoa. Immunofluorent staining and flow cytometry showed that the expression of gH2AX was increased in a dose-dependent and time-dependant manner after the treatment of adriamycin. Adriamycin also induced the concurrent appearance of DNA maintenance/repair proteins RAD50 and 53BP1 with gammaH2AX in spermatozoa. Wortmannin, an inhibitor of the phosphatidylinositol 3-kinase (PI3K) family, abolished the co-appearance of these two proteins with gammaH2AX.</p><p><b>CONCLUSION</b>Human mature spermatozoa have the same response to DSB-induced H2AX phosphorylation and subsequent recruitment of DNA maintenance/repair proteins as somatic cells.</p>


Sujets)
Humains , Mâle , Androstadiènes , Pharmacologie , Antibiotiques antinéoplasiques , Pharmacologie , Cellules cultivées , Test des comètes , Cassures double-brin de l'ADN , Altération de l'ADN , Enzymes de réparation de l'ADN , Métabolisme , Protéines de liaison à l'ADN , Métabolisme , Doxorubicine , Pharmacologie , Interactions médicamenteuses , Cytométrie en flux , Histone , Métabolisme , Protéines et peptides de signalisation intracellulaire , Métabolisme , Phosphorylation , Inhibiteurs de protéines kinases , Pharmacologie , Spermatozoïdes , Biologie cellulaire , Métabolisme , Protéine-1 liant le suppresseur de tumeur p53
5.
Journal of Zhejiang University. Medical sciences ; (6): 439-442, 2007.
Article Dans Chinois | WPRIM | ID: wpr-271506

Résumé

<p><b>OBJECTIVE</b>To evaluate the levels of bone morphogenetic protein-15 (BMP-15) in human follicular fluid (FF) and its association with response to ovarian stimulation.</p><p><b>METHODS</b>Western blotting was performed to determine the levels of BMP-15 in FF obtained from follicle aspirates in 70 patients undergoing IVF treatment. According to the response to ovarian stimulation the patients were divided into poor responder group and normal responder group.</p><p><b>RESULT</b>BMP-15 levels in FF of poor responders were significantly higher than those in normal responders (1.01 +/- 0.34 vs 0.77 +/- 0.24, P<0.01).</p><p><b>CONCLUSION</b>Increased levels of BMP-15 in FF may be associated with poor response to ovarian stimulation.</p>


Sujets)
Adulte , Femelle , Humains , Technique de Western , Protéine morphogénétique osseuse de type 15 , Hormone folliculostimulante , Liquide folliculaire , Métabolisme , Hormone de libération des gonadotrophines , Facteur-9 de croissance et de différenciation , Infertilité féminine , Métabolisme , Protéines et peptides de signalisation intercellulaire , Ovaire , Métabolisme , Induction d'ovulation
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