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AIM:To investigate the effect of arsenic trioxide (ATO) on T-bet/GATA3 signal pathway in MRL/lpr mice.METHODS:MRL/lpr mice and C57BL/6J mice at the age of 20 weeks were chosen and then divided in 2 different sub-groups,respectively.The mice in 2 sub-groups received ATO (0.4 mg · kg-1.d-1) and sodium chloride (NS,volume weight-determined) by intraperitoneal injection respectively for 2 months.Afterward,the spleens were isolated fron the MRL/lpr and C57BL/6J mice under pathogen-free condition and the suspensions were prepared.The mRNA level of T-bet,GATA3,IFN-γ,IL-4 and the mRNA ratio of T-bet/GATA3 were detected by RT-qPCR.The protein expression of T-bet and GATA3 was determined by Western blot.The serum levels of IFN-γ and IL-4 were measured by ELISA.RESULTS:The mRNA and protein levels of T-bet,IFN-γand the mRNA ratio of T-bet/GATA3 in NS group of MRL/lpr mice were higher than those in NS group of C57BL/6J mice (P ＜0.05).However,the GATA3 and IL-4 were lower in NS group of MRL/lpr mice in both mRNA and protein level (P ＜ 0.05).In MRL/lpr mice,the mRNA and protein levels of T-bet,IFN-γ and the mRNA ratio of T-bet/GATA3 were lower in ATO group compared with NS group (P ＜ 0.05),no difference was found in GATA3 and IL-4.No difference of the indexes mentioned above between ATO group and NS group in C57BL/6J mice was observed.CONCLUSION:ATO may affect the signaling pathway of T-bet/GATA3 to down-regulate the mRNA expression and the protein secretion of IFN-γ by decreasing the expression of T-bet in MRL/lpr mice.
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AIM:To investigate the effect of arsenic trioxide (ATO) on T-bet/GATA3 signal pathway in MRL/lpr mice.METHODS:MRL/lpr mice and C57BL/6J mice at the age of 20 weeks were chosen and then divided in 2 different sub-groups,respectively.The mice in 2 sub-groups received ATO (0.4 mg · kg-1.d-1) and sodium chloride (NS,volume weight-determined) by intraperitoneal injection respectively for 2 months.Afterward,the spleens were isolated fron the MRL/lpr and C57BL/6J mice under pathogen-free condition and the suspensions were prepared.The mRNA level of T-bet,GATA3,IFN-γ,IL-4 and the mRNA ratio of T-bet/GATA3 were detected by RT-qPCR.The protein expression of T-bet and GATA3 was determined by Western blot.The serum levels of IFN-γ and IL-4 were measured by ELISA.RESULTS:The mRNA and protein levels of T-bet,IFN-γand the mRNA ratio of T-bet/GATA3 in NS group of MRL/lpr mice were higher than those in NS group of C57BL/6J mice (P ＜0.05).However,the GATA3 and IL-4 were lower in NS group of MRL/lpr mice in both mRNA and protein level (P ＜ 0.05).In MRL/lpr mice,the mRNA and protein levels of T-bet,IFN-γ and the mRNA ratio of T-bet/GATA3 were lower in ATO group compared with NS group (P ＜ 0.05),no difference was found in GATA3 and IL-4.No difference of the indexes mentioned above between ATO group and NS group in C57BL/6J mice was observed.CONCLUSION:ATO may affect the signaling pathway of T-bet/GATA3 to down-regulate the mRNA expression and the protein secretion of IFN-γ by decreasing the expression of T-bet in MRL/lpr mice.
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Aim To explore the protective effects and underlying mechanisms of Liu weiwuling Tablets (LW-WL)in concanavalin A (ConA)induced acute immu-nological liver injury in mice.Methods Mice were randomly divided into control,model,Bicyclol,LW-WL low dose (8 g·kg-1 )and LWWL high dose (16 g ·kg-1 )group.The medicattion was performed once daily for seven consecutive days,then the model of im-munological liver injury was prepared by intravenous injection of ConA (15mg·kg-1)in the tail of mice in each group except for the control group one hour after the last treatment.The pathological changes of liver tissues of mice were evaluated by HE staining with, and the levels of alanine amino transferase (ALT),as-partate aminotransferase (AST),and total bilirubin (TBIL)in serum were analyzed by colorimetric meth-od;the level of interleukin 12 (IL-12 ),interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α),interleu-kin 4 (IL-4)and interleukin 10 (IL-10)in liver was measured by real-time quantitative polymerase chain reaction (RT-qPCR);the changes of Th1 (IFN-γ) and Th2 (IL-4)cells were observed by flow cytometric (FCM)analysis;the expression of Th1/Th2 transcrip-tion factor T-bet/GATA-3 in liver tissue was detected by Western blot.Results Compared with normal con-trol group,the serum ALT,AST and TBIL were signif-icantly increased in model group, the pathological damage of the liver tissue was severe,and the necrosis and apoptosis of hepatic cells were large, which showed that the model was successful .Compared with model group,both low and high dose of LWWL could significantly reduce ALT,AST,TBIL levels in serum induced by ConA;Th1 cells in the spleen decreased, while Th2 cells increased;the expressions of IL-12, IFN-γand TNF-αmRNA were significantly inhibited with IL-4 and IL-10 mRNA expression elevated in mouse liver tissue;the expression of GATA-3 protein was up-regulated,T-bet protein expression showing no significant changes.Conclusion LWWL could regu-late Th1/Th2 balance,thus inhibiting the acute immu-nity hepatic injury induced by ConA.
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Objective:To discuss the influence of dexamethasone(DXM) to T-bet/GATA-3 of athma rats.Methods: 30 SD rats were randomly divided into normal group,model group and DXM group,10 rats in each group.Ovalbumin was intraperitoneal injected on the 1th and 8th day and aerosol inhaled from the 15th day,once a day,14 days altogether.Dexamethasone was intraperitoneal injected in 0.5 mg/kg from the 15th day,once per day,a total of 14 times.Flow cytometry tested the content of Th1 and Th2 in spleen and peripheral blood,immunohistochemistry and Western blot measured T-bet and GATA-3 protein expression and RT-PCR tested T-bet and GATA-3 mRNA expression.Results: Compared model group with normal group,the contents of Th1,Th2 in spleen and peripheral blood had a very significant increase(P<0.01);expression of T-bet,GATA-3 protein and mRNA in the lung tissue also had very significant risen(P<0.01).While compared DXM group with model group,T-bet/GATA-3 rose significantly(P<0.05)or very significantly(P<0.01).Conclusion: T-bet/GATA-3 is the key transcription factors that influence the balance of Th1/Th2,while DXM can raise the ratio of T-bet/GATA-3,which reduce the severity of asthma.
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Objective To investigate the effects of Lipo-PGE1 on the expression of T-bet and Gata-3,and its potential mechanisms causing the shift of T cells from Th1 to Th2 on Acute lung injury(ALI)induced by Lipopolysaccharide(LPS)in mice.Methods Sixty male BALB-C mice were randomly divided into three groups(n =20 in each group):(1)control group,mice were treated with intravenous injection of NS in dose of 10 ml/kg,(2)LPS group,mice were exposed to LPS with dosage of 5 mg/kg(0.5 g/ml diluted in saline),and(3)LPS + PGE1 group,mice were treated with Lipo-PGE1 in dose of 15μg/kg.Sixhours after injection,the lungs were removed for observing the histopathological changes and determination of wet/dry lung weight(W/D)ratio.The levels of Th1 and Th2 were determined by flow cytometry,and the expressions of T-bet and Gata-3 mRNA were detected by using RT-PCR.One-way ANOVA was used for comparing differences between groups,and all data were presented in((x)± s).Results The histological changes of lung injury were lessened by PGEC ompared with the W/D ratio(5.74 ± 0.31)in LPS group,the one(4.92 ±0.27)in LPS +PGE1 group was lower significantly(P ＜0.01).The levels of Th1 and Th2 and their ratio Were higher in LPS +PGE1 group[(20.31 ±2.20)％,(10.50±0.80)％,(1.93±0.05)]than in LPS group[(16.65 ±1.70)％,(9.40 ±1.25)％,(1.73 ±0.03)](P＜0.01).Compared with control group,the expressions ofT-bet mRNA(1.183 ±0.495),and Gata-3 mRNA(0.693±0.285),and their ratio(1.713 ± 0.131)were lower(P ＜0.01); compared with LPS group,PGE1 significantly increased the expressions of T-bet mRNA(1.827 ± 0.705)and the ratio of T-bet/Gata-3 (2.502 ±0.352)(P ＜0.01),while didn(t)increased the expressions of Gata-3 mRNA(0.7191 ±0.186)significantly(P ＞ 0.05).Conclusions Lipo-PGE1 may up-regulate transcription factor T-bet which participates in the Th1 differentiation ratio,and then improve the inflammatorv svmntom.