ABSTRACT
Objective To isolate and culture rats liver KCS ,and to explore the effect of IRE1-XBP1 pathway on regulation of polarization in activated Kupffer cells (KCs) .Methods (1)Rat KCs were isolated by Ⅳ type collagenase digestion and gradient cen-trifugation methods .(2)KCs were transfected and randomly divided into four groups :XBP1-shRNA group ,Ctrl-shRNA group , AdV-XBP1 group and Ctrl-AdV group .(3)The transfection level of KCs XBP1 ,IL-6 ,IFN-γ ,TNF-α and IL-17 were detected by RT-PCR ;the protein expression level of JAK1 ,JAK2 ,STAT1 and STAT3 were evaluated by Western blot .(4)The changes of KCs expression type in each group were detected by flow cytometry (FCM ) and the laser confocal .(5)T cells derived from rat spleen cells were co-cultured within the 4 groups of KCs mentioned above ;T cells proliferation was measured by Brdu labeling assay .(6)T cells apoptosis was determined by Annexin V /PI FCM analysis .(7)The density of IL-6 ,IFN-γ ,TNF-α ,IL-17 and IL-10 in the su-pernatant of co culture was assessed by ELISA .Results (1)The mRNA and protein level of XBP1 were measured by RT-PCR and western blot ,those in XBP1-shRNA group were significantly reduced compared with those in the other three groups ,while in AdV-XBP1 groups ,results demonstrated entirely the opposite tendency (P< 0 .05) .(2)The expression of marker molecules on the sur-face of KCs such as M HC Ⅱ ,CD86 and CD40 in XBP1-shRNA group were significantly lower (P< 0 .05) ,but CD204 and CD206 expression were much higher compared with the other three (P< 0 .05) .However the expression tendency of these surface markers were shown the opposite results in AdV-XBP1 group (P < 0 .05) .(3)Western blot revealed the XBP1-shRNA could statistically suppress the protein levels and phosphorylation of JAK 1 ,JAK2 ,STAT1 and STAT3 ,which involved in the pro inflammatory cyto-kines regulation and KCs polarization (P< 0 .05) .But in AdV-XBP1 group ,these protein and its phosphorylation were markedly promoted (P< 0 .05) .ELISA results collaborated with Western blot .(4)3 d after co cultured with KCs transfected with XBP1-shR-NA ,the levels of T lymphocyte proliferation and pro inflammatory cytokines secretion were significantly reduced ,but the levels of T lymphocyte apoptosis and anti inflammatory cytokines secretion were remarkably enhanced (P< 0 .05) .Conclusion Blockage of IRE1-XBP1 activation could alter the phenotype of active KCs to M 2 like type and attenuated the capacity of antigen present of KCs ,while up regulated the expression of IRE1-XBP1 pathway could change the phenotype of KCs to M 1 type plus the promotion of antigen present capacity .
ABSTRACT
Objective To discuss the short-term results of tricuspid valve annuloplasty with prosthetic ring.Methods Tricuspid valve annuloplasty with prosthetic ring was performed in 34 patients with organic or functionaltricuspid valve insufficiency. At the same time atrial septal defect and ventrieular septal defect were closed valve re-placement was done. Results There was no in-hospital death. Echocardiography was done about one week after oper-ations,which showed that fight ventricle or pulmonary artery were smaller than that before operation in all patients andresidual tricuspid valve insufficiency was found mild in 11 cases, moderate in 2 cases. Conclusion Tricuspid valveannuloplnsy should be performed in patients with organic or functional tricuspid valve insufficiency. Better short-termresults may be got when prosthetic ring was used to small tricuspid annulus.