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National Journal of Andrology ; (12): 687-691, 2017.
Article in Chinese | WPRIM | ID: wpr-812895


Objective@#To investigate the influence of deep slow-wave sleep deprivation on the oxidative stress of testicular tissue in rats.@*METHODS@#Thirty-six 5-week-old male Wistar rats were equally randomized into deep slow-wave sleep deprivation group (SD1), deep slow-wave sleep and duration sleep deprivation group ( SD2), and a cage control group (CC). The rat model of deep slow-wave sleep deprivation was established using the flowerpot technique. The rats in the SD1 group were interfered every 24 minutes and deprived of 12 hours of sleep at night, those in the SD2 group deprived of 8 minutes of sleep at an interval of 24 minutes and 12 hours of sleep at night, and those in the CC group exposed to 12 hours of daylight and 12 hours of darkness. After 28 days, all the rats were executed for measurement of the testis volume and protein content, determination of the methane dicarboxylic aldehyde (MDA) level and activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and observation of the pathological changes in the testicular tissue under the microscope.@*RESULTS@#Compared with the CC group, the rats in the SD1 and SD2 groups showed significantly reduced body weight ([268.5 ± 1.6] vs [248.1 ± 25.1]and[232.9 ± 10.1]g, P0.05). The lumens in the testis were narrowed, with obvious hyperplasia, hyperemia and edema in the peripheral interstitial tissue, but no significant pathologic changes were observed in the testis tissue of the SD1 group.@*CONCLUSIONS@#Long-term deprivation of deep slow-wave sleep impairs the structure of the testis tissue and induces oxidative stress response in rats.

Animals , Body Weight , Glutathione Peroxidase , Male , Malondialdehyde , Oxidative Stress , Random Allocation , Rats , Rats, Wistar , Sleep Deprivation , Metabolism , Sleep Stages , Superoxide Dismutase , Testis , Metabolism , Pathology , Time Factors , Weight Loss
Chinese Medical Journal ; (24): 942-948, 2010.
Article in English | WPRIM | ID: wpr-242541


<p><b>BACKGROUND</b>Rapamycin (RAPA) is a relatively new immunosuppressant drug that functions as a serine/threonine kinase inhibitor to prevent rejection in organ transplantation. RAPA blocks activation of T-effector (Teff) cells by inhibiting the response to interleukin-2. Recently, RAPA was also shown to selectively expand the T-regulator (Treg) cell population. To date, no studies have examined the mechanism by which RAPA converts Teff cells to Treg cells.</p><p><b>METHODS</b>Peripheral CD4(+)CD25(-) naive T cells were cultivated with RAPA and B cells as antigen-presenting cells (APCs) in vitro. CD4(+)CD25(-) T cells were harvested after 6 days and analyzed for expression of forkhead box protein 3 (Foxp3) using flow cytometry. CD4(+)CD25(+)CD127(-) subsets as the converted Tregs were isolated from the mixed lymphocyte reactions (MLR) with CD127 negative selection, followed by CD4 and CD25 positive selection using microbeads and magnetic separation column (MSC). Moreover, mRNA was extracted from converted Tregs and C57BL/6 naive CD4(+)CD25(+) T cells and Foxp3 levels were examined by quantitative real-time polymerase chain reaction (rt-PCR). A total of 1 x 10(5) carboxyfluorescein succinimidyl ester (CFSE)-labeled naive CD4(+)CD25(-) T cells/well from C57BL/6 mice were cocultured with DBA/2 or C3H maturation of dendritic cells (mDCs) (0.25 x 10(5)/well) in 96-well round-bottom plates for 6 days. Then 1 x 10(5) or 0.25 x 10(5) converted Treg cells were added to every well as regulatory cells. Cells were harvested after 6 days of culture and analyzed for proliferation of CFSE-labeled naive CD4(+)CD25(-) T cells using flow cytometry. Data were analyzed using CellQuest software.</p><p><b>RESULTS</b>We found that RAPA can convert peripheral CD4(+)CD25(-) naive T Cells to CD4(+)Foxp3(+) Treg cells using B cells as APCs, and this subtype of Treg can potently suppress Teff proliferation and maintain antigenic specificity.</p><p><b>CONCLUSION</b>Our findings provide evidence that RAPA induces Treg cell conversion from Teff cells and uncovers an additional mechanism for tolerance induction by RAPA.</p>

Animals , Antibiotics, Antineoplastic , Pharmacology , Antigen-Presenting Cells , Allergy and Immunology , Metabolism , B-Lymphocytes , Allergy and Immunology , Metabolism , CD4-Positive T-Lymphocytes , Allergy and Immunology , Metabolism , Cell Proliferation , Dendritic Cells , Allergy and Immunology , Metabolism , Forkhead Transcription Factors , Metabolism , Interleukin-2 Receptor alpha Subunit , Metabolism , Interleukin-7 Receptor alpha Subunit , Metabolism , Male , Mice , Mice, Inbred C57BL , Mice, Inbred DBA , Mitomycin , Pharmacology , Polymerase Chain Reaction , Sirolimus , Pharmacology , T-Lymphocytes, Regulatory , Allergy and Immunology , Metabolism
Article in Chinese | WPRIM | ID: wpr-270682


<p><b>OBJECTIVE</b>To study the effect of rapamycin in inducing naïve murine effector T cell (Teff) convert to regulatory T cell (Treg) in vitro.</p><p><b>METHODS</b>The forkhead box protein 3 (FoxP3) negative Teff were isolated and purified from the spleen and lymph node of C57 BL/6 murines aged 6-8 weeks, then Teff were cultured in three groups with mature dendritic cells (mDC), B cells, and plate coated Anti-CD3. In addition, the control wells and the test wells were prepared in each group, rapamycin were not added in the control wells but added in the test wells with concentrations of 1, 10, 50, and 100 nmol/L. Percentages of FoxP3 positive Treg were examined by flow cytometry after 4 days in Anti-CD3 group and after 6 days in the other two groups.</p><p><b>RESULTS</b>As shown by the flow cytometry, the percentages of FoxP3 positive Treg were as follows in three group: in the mDC group, it was 0.01% in the control well and 0.39%, 0.47%, 0.34%, and 0.26% in test wells; in B cell group, it was 0.01% in the control wells and 5.56%, 5.89%, 7.15%, and 4.72% in the test wells; in Anti-CD3 group, it was 0.93% in the control wells and 1.35%, 1.07%, 1.02%, and 1.19% in test wells. No significant difference was found between the test wells and control wells in the mDC group and Anti-CD3 group; however, the percentages of FoxP3 positive Treg was significantly different between the test wells and control wells in the B cell group (P < 0.01).</p><p><b>CONCLUSION</b>When B cell is acted as the antigen-presenting cell, rapamycin can effectively induce Teff convert to Treg in vitro.</p>

Animals , B-Lymphocytes , Cell Biology , Allergy and Immunology , Cell Differentiation , Cells, Cultured , Dendritic Cells , Cell Biology , Allergy and Immunology , Flow Cytometry , Forkhead Transcription Factors , Allergy and Immunology , Mice , Mice, Inbred C57BL , Mice, Inbred DBA , Precursor Cells, T-Lymphoid , Cell Biology , Allergy and Immunology , Sirolimus , Pharmacology , T-Lymphocyte Subsets , Cell Biology , Allergy and Immunology , T-Lymphocytes, Regulatory , Cell Biology , Allergy and Immunology