Effect Of STAT5 Gene Silencing On The Proliferation Of T Lymphocytes In A Mice Model Of Asthma

Objective:Silence STAT5a/b genes of T lymphocytes in vitro and discuss the effect of silencing STAT5 gene on the proliferation function of T lymphocytes. Provide a theoretical basis for clarifying the pathogenesis of asthma furtherly, and look for more targeted treatment methods.Methods:BALB/c mice were sensitized with ovalbumin through Intraperitoneal injection, repeated challenged with exposure to aerosolized ovalbumin. Normal and asthma spleen T lymphocytes were selected by immunomagnetic bead, and the STAT5a/b genes of these T-lymphocytes were silenced by siRNA. We detected the mRNA of STAT5a/b by Fluorescence quantitative PCR, measured the protein by WESTERN BLOT, detected the proliferation rates of T-lymphocytes by CCK-8, and estimated the cell cycle and cell apoptosis by flow cytometry.Results:(1) The inflammatory cell infiltration was present in asthma mice airway, while no significant infiltration of inflammatory cells in normal mice. Mouse alveolar lavage fluid cell count showed there were more eosinophils, lymphocytes, heterophil and granulocytes compared with normal mice.(2) The STAT5a/b expression of asthmatic group was significantly higher compared with normal group. (3) The results of REAL TIME PCR, and WESTERN BLOT showed that the mRNA and protein expression of STAT5a/b were significantly reduced after siRNA-specific intervention compared with blank control group and missense-chain group. (4)The result of CCK-8 tests showed that T lymphocytes proliferation rates were significantly reduced after siRNA-specific intervention compared with the blank control group group and the missense-chain group. (5)Flow cytometry showed the T-lymphocytes in proliferative phase were significantly reduced after siRNA-specific intervention compared with the blank control group and the missense-chain group. (6)The apoptosis rates of T lymphocytes were significantly increased after siRNA-specific intervention compared with blank control group and missense-chain group. Conclusion:STAT5a/b is an imr(?)ant transcription factor which mediates T lymphocyte proliferation. The expression of STAT5a/b in asthmatic mouse is higher compared with normal mice. RNA interference can specifically reduce the expression of STAT5a/b, inhibit the proliferation of T lymphocytes, and promote the apoptosis of T lymphocytes. STAT5a/b is expected as a new target for the treatment of asthma.