Effect of T cell immunoglobulin domain and mucin domain protein-3 gene interfered by short hairpin RNA on helper T lymphocytes 17 differentiation and proinflammatory capacity in asthmatic mice / 中华实用儿科临床杂志

ABSTRACT

Objective To investigate the effect of down-regulating of T cell immunoglobulin domain and mucin domain protein-3 (Tim-3) gene in asthmatic mouse model by short hairpin RNA(shRNA) and explore the role of Tim-3 on the differentiation of helper T lymphocytes 17 (Thl7),airway inflammation,as well as airway hyperresponsiveness in pathogenesis of asthma.Methods An asthmatic murine model was established by way of ovalbumin (OVA) sensitization and challenge.Treating Tim-3 gene was intranasally administered with Tim-3-specific shRNA.Invasive pulmonary impedance method was adopted to detect mice airway resistance.Flow cytometry analysis was performed to determine the levels of Th17 ; enzyme linked immunosorbent assay was performed to determine the concentrations of IL-17 and transforming growth factor-beta(TGF-β) in supematant.Results Asthmatic mice model was successfully established.By using Tim-3 shRNA silencing Tim-3,airway inflammation was reduced and airway hyperresponsiveness was declined in asthmatic group ;the levels of Th17 cells in asthmatic group [(6.43 ± 1.01)％] were significantly increased compared with normal controls[(1.75 ± 0.02) ％].After using Tim-3 shRNA silencing Tim-3,the levels of Th17 cells [(2.36 ± 0.28) ％] were decreased (F =40.05,P ＜ 0.05) ; the level of IL-17 in asthmatic group [(118.8 ± 16.5) ng/L] was significantly increased compared with normal controls[(72.5 ± 13.6)ng/L],after using Tim-3 shRNA silencing Tim-3,the level of IL-17 [(73.6 ± 12.5) ng/L] was decreased (F =32.80,P ＜ 0.05),while the expression of TGF-β did not change.Conclusions Down regulation of Tim-3 gene can decrease airway inflammation and airway hyperresponsiveness,which may be related to the change of Th17 cell differentiation.