ABSTRACT
Objective To investigate the role of miR-10a in CD4+CD25+Treg-mediated immunosuppression during sepsis and its potential role in immunotherapy for sepsis.Methods Sepsis mouse model was established by cecal ligation and puncture(CLP).Balb/c mice of clean grade were sacrificed 1,3,5,and 7 days after operation.Blood as well as spleen samples were harvested at given intervals.The splenic CD4+CD25+Treg cells and CD4+T cells were isolated by MACS microbeads.Cells were cultured,and phenotypes were analyzed by flow cytometry.The miR-10a expressed in Treg cells were detected by Real-time PCR.After administration of LV-mmu-miR-10a-5p-inhibition,the immunosuppressive function have been detected.Statistical analyses were performed using one-way analysis of variance (SPSS 19.0,Chicago,USA) test followed by Dunnett-t test to compare among three or more groups or by Student's t-test to compare between two groups.Results The percentages of splenic Tregs (CD4+CD25+/CD4+T) was (7.34±1.2)％ in normal group,and the increase in percentage of Tregs in spleen has been observed in septic mice (P＜0.05).The mean fluorescence intensity (MFI) of Foxp3+Treg was increased in septic mice compared with sham group (P＜0.05).The expression of miR-10a was significantly elevated on CLP 1-7 day (P＜0.05).After down-regulation of miR-10a in septic mice,the percentages of Tregs (CD4+CD25+/CD4+T) was significantly increased in septic mice (P＜0.05),the MFI of Foxp3+Treg was increased in septic mice compared with control group (P＜0.05).The CD4+T cell proliferative activity in CLP-induced mice was significantly suppressed on CLP 3 day compared with sham group (P＜0.05).After down-regulation of miR-10a in septic mice,the CD4+T cell proliferative activity was significantly suppressed compared with control group (P＜0.05).Conclusions Treg plays a critical role in immunosuppression in septic mice.Inhibition of miR-10a in vivo could enhence immunesuppression of CD4+CD25+Treg.Therefore miR-10a may participate in the regulation of CD4+CD25+Treg immunosuppression in sepsis and become the target for immunotherapy.
ABSTRACT
Objective To explore the relationship between galactose-induced apoptosis of lens epithelial cells and cataractogenesis.Methods Galactose was injected retrobulbarly into Wistar rats.The opacity of lens was examined by slit lamp.The expression of C-MYC was measured by flow cytometer.The apoptosis of lens epithelial cells was observed by TUNEL techniques.Results More than 50 percent of lenses were in the Ⅱ stage of cataractogenesis on the(7 d) after galactose induction;Eighty-seven percent of lenses were in the Ⅳ-Ⅴ stage on the(14 d),and 100 percent of lenses were in the Ⅳ-Ⅴ stage on the(24 d).The rate of apoptotic cell on the(7 d) and on the(14 d) after galactose induction was respectively 5.6%～8.4% and 30.2%～41.8%,and 60% apoptotic cells were observed on the(24 d).The levels of C-MYC expression in lens epithelial cells induced by galactose on the(7 d) and(14 d) were higher than that of normal lenses,but the level of C-MYC expression backed to normal level on the(24 d).Conclusion It was suggested that there are some apoptosis of the lens epithelial cells during galactose-induced cataractogenesis and that the apoptosis of the lens epithelial cells might be caused by higher expression of C-MYC.