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Background: The etiology and pathogenesis of irritable bowel syndrome (IBS) are not yet clear, and is lack of effective means for its prevention, diagnosis and treatment in clinic. In recent years, more and more studies showed that intestinal local mucosal immune dysfunction plays an important role in the pathogenesis of IBS. Aims: To investigate the role of intestinal dendritic cells (DC) in the imbalance of colonic mucosal Th1/Th2 immune response pathway in IBS with diarrhea (IBS-D) rats and its effect on visceral hypersensitivity. Methods: Forty-five adult SD rats were randomly divided into blank control group, NS control group and model group, and each group consisted of 15 rats. Visceral hypersensitive model was established by acetic acid enema combined with restraint stress. Abdominal withdrawal reflex (AWR) test and fecal characteristics were used to evaluate the visceral sensitivity. Flow cytometry separation technique was used to isolate DC from mesenteric lymph nodes and CD4
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Objective To study the changes of Th1 and Th2 cells in peripheral blood of patients with polycystic ovary syndrome(PCOS),and to explore the significance of Th1/Th2 imbalance in the pathogenesis of PCOS.Methods 41 patients who were diagnosed with PCOS were selected as PCOS group,and 25 healthy women were set as the control group.Peripheral venous blood(8 ml) were obtained from subjects in the two groups,serum was separated,and chemiluminescence was used to detect sexual hormone levels.Peripheral blood mononuclear cells (PBMCs) were obtained,cytokines IFN-gamma and IL-4 were detected by ELISA,and Th1 and Th2 cells were detected by flow cytometry.The data were statistically analyzed,with P＜0.05 as the difference being statistically significant.Results ELISA results showed that compared with the control group,there was no significant change in the level of cytokine IL-4 in the peripheral blood of the PCOS group (P＞0.05),while the level of IFN-gamma increased significantly (the control group:(54.2±18.7) pg/ml,the PCOS group:(136.8±24.6) pg/ml,P＜0.05).The results of flow cytometry showed there was no significant change in the proportion of Th2 cells(P＞0.05),while the proportion of Th1 cells (the control group:(5.7±0.6)％,PCOS group:(14.3t±1.1)％,P＜0.05) and the ratio of Th1/Th2 increased significantly (the control group:2.1 ±0.3,PCOS group:4.4±0.5,P＜0.05).Correlation analysis showed that Th1/Th2 ratio in PCOS group was positively correlated with LH (r=0.512,P=0.034),LH/FSH (r=0.584,P=0.028) and T(r=0.631,P=0.016).Conclusion Th1/Th2 imbalance was found in PCOS patients,which may be closely related to the disorder of sexual hormone levels.
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ObjectiveTo examine whether Shenfu injection (SFI) reduces post-resuscitation myocardial dysfunction in a pig model by modulating expression imbalance of transcription factors of regulatory T cell, namely GATA-3 and T-bet.Methods Thirty pigs were randomly divided into sham group (n = 6) and cardiopulmonary resuscitation (CPR) group (n = 24) according to the random number table method, and the pigs in the CPR group were randomly subdivided into normal saline (NS) group, epinephrine (EP) group, and SFI group (n = 8 per group). After 8minutes of untreated ventricular fibrillation (VF) followed by 2 minutes of CPR, animals in three groups respectively received central venous injection of either 20 mL SFI (1.0 mL/kg, SFI group), EP (0.02 mg/kg, EP group) or NS (NS group). Blood samples were obtained before VF and 0.5, 2, 6 hours after restoration of spontaneous circulation (ROSC), and the parameters of hemodynamics and oxygen metabolism were determined. Surviving pigs were sacrificed at 24 hours after ROSC, the pathological changes in myocardium were observed, the levels of interleukin-4 (IL-4), tumor necrosis factor-α (TNF-α) andγ-interferon (IFN-γ) were measured by enzyme linked immunosorbent assay (ELISA), and expressions of protein and mRNA of GATA-3 and T-bet were determined by Western Blot and quantitative real-time polymerase chain reaction (RT-qPCR), respectively.Results Six pigs of three resuscitation groups were successfully resuscitated. The CPR time, number of defibrillation, defibrillation energy, and ROSC time were significantly decreased in the EP and SFI groups compared with those in the NS group. Compared with the sham group, the parameters of left ventricular systolic function and oxygen metabolism were significantly decreased, myofibril organelles were extensively damaged, and progressive and severe deterioration of the myocardium was found, and mitochondrial structure was not recognizable in the NS group; the level of IL-4 in myocardium were markedly decreased, while that of TNF-α, IFN-γand IFN-γ/ IL-4 [reflecting helper T cell 1/2 (Th1/Th2)] were significantly increased. Protein and mRNA expressions of GATA-3 were markedly reduced in the myocardium of pigs in the NS group compared with that of the sham group at 24 hours after ROSC, while T-bet was significantly increased. Compared with the NS group, animals treated with SFI had minimal myocardial intracellular damage, with decreased heart rate (HR, bpm: 90.33±3.79 vs. 106.83±5.36) and increased mean arterial pressure (MAP), cardiac output (CO), oxygen delivery (DO2), and oxygen consumption (VO2) at 6 hours after ROSC [MAP (mmHg, 1 mmHg = 0.133 kPa): 107.67±1.96 vs. 86.83±1.85, CO (L/min): 2.47±0.08 vs. 2.09±0.04, DO2 (mL/min): 364.31±4.21 vs. 272.33±3.29, VO2 (mL/min): 95.00±2.22 vs. 82.50±2.28, allP<0.05]. Compared with the NS groups at 24 hours after ROSC, level of IL-4 was markedly increased in myocardial cells (ng/L: 33.80±3.06 vs. 16.15±1.34,P< 0.05), while the levels of TNF-α, IFN-γ and IFN-γ/IL-4 were lowered significantly [TNF-α (ng/L): 18.16±0.71 vs. 29.64±1.89, IFN-γ (ng/L): 373.75±18.36 vs. 512.86±27.86, IFN-γ/IL-4: 16.15±1.34 vs. 33.80±3.06, allP< 0.05], and myocardial T-bet protein and mRNA expressions were reduced [T-bet protein (gray value): 0.41±0.07 vs. 0.59±0.11, T-bet mRNA (2-ΔΔCt): 4.37±0.21 vs. 7.57±0.55, bothP< 0.05], furthermore, myocardial GATA-3 protein and mRNA expressions were significantly up-regulated in SFI group [GATA-3 protein (gray value): 0.25±0.07 vs. 0.16±0.07, GATA-3 mRNA (2-ΔΔCt): 0.63±0.07 vs. 0.34±0.05, bothP< 0.05]. The parameters in SFI group were significantly improved compared with those of the EP group.ConclusionsMyocardial immune dysfunction is induced by Th1/Th2 imbalance following myocardial injury subsequent to CPR in pigs. SFI can attenuate myocardial injury and regulate myocardial immune disorders, protect post-resuscitation myocardial injury by modulating expression imbalance of transcription factors GATA-3 and T-bet.
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Objective To study the role of imbalance between transcription factors GATA-3 and T-bet expressions in causing acute lung injury after resuscitation in cardiac arrest model of swine.Methods Mter swine model of electrically induced cardiac arrest was established for 8 minutes,animals were resuscitated to get restoration of spontaneous circulation (ROSC).The swine with ROSC were randomly assigned to be sacrificed at 12 and 24 h after ROSC (n =8 in each group).CD3 +,CD4+ and CD8 + lymphocyte subsets were determined by flow cytometry,and the levels of serum IL-4,TNF-α,and IFN-γ were measured by using ELLSA.The protein levels and expressions of GATA-3/T-bet mRNA were detected in lung tissue by western blotting and quantitative real-time PCR device,respectively.Results Pulmonary function was significantly impaired after ROSC.CD4 + lymphocyte subsets (28.4 ± 2.3) ％,(24.1 ± 1.6) ％ and CD4 +/CD8 + (1.7 ±0.9),(1.5 ± 1.0) were significantly lower in the post-ROSC group compared with the sham-operated group (48.4±2.9)％,(51.1±5.4)％ (2.5±1.3),(2.7±1.1) (P＜0.05) at 12 h and 24 h after ROSC.The levels of serum IL-4 and TNF-α were markedly increased,while IFN-γ and IFN-γ/IL-4 were significantly decreased in the post-ROSC group compared with the sham-operated group (P ＜0.05) at 2-12 h after ROSC.Protein level and expression of GATA-3 mRNA in lung tissue were markedly increased,while those of T-bet were significantly reduced in the post-ROSC group compared with the sham-operated group (P ＜0.05) at 12 and 24 h after ROSC.Conclusions The lung immune dysfunction induced by imbalance between transcription factors GATA-3 mRNA and T-bet mRNA expressions may complicate in the process of post-resuscitation lung injury in a porcine model of cardiac arrest.
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Objective To investigate the role of basophils in the imbalance of Th 1/Th2 response in mouse models of collagen-induced arthritis(CIA).Methods 4-6-weeks old C57/BL6 mice were immunized with collagen at multiple points on the back and foot twice (0 and 3 weeks) to establish a mouse model of collagen-induced arthritis.Blood samples were collected before the first immunization and 1, 3, 6 and 9 weeks after immunization , and cells from lymph nodes were collected.Flow cytometry and ELISA were employed to detect the levels of basophils and IL-4, and the joint swelling was scored.Results Mouse model of CIA was successful established .The ratio of IL-4/IFN-γof the CIA group was significantly lower than that in the mice before CIA modeling and the control group , indicating a Th2-dominant response .At the same time, the peripheral basophils counting and percentage of IL-4 positive basophils of the CIA group were significantly higher than those of the control group .While, the IL-4/IFN-γratio of the CIA group was significantly higher than that of the control group , indicating a Th1-dominant response .The peripheral basophils counting of the CIA group was slightly lower than that of the control group .Conclusion Basophils may participate in the development of CIA in mouse models through affecting the imbalance of Th 1/Th2 response.
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AIM: To explore the effect of liquorice on airway inflammation and Th_1/Th_2 imbalance in chronic asthma. METHODS: The chronic asthma model was established by intraperitoneal ovalbumin. The effect of liquorice on mice model of chronic bronchial asthma was observed, and the levels of serum IFN-? and IL-4 were tested by ELISA. RESULTS: Decreasing inflammatary cells infiltration in pulmonary tissue slices of tiny bronchial wall together with increasing serum IFN-? and decreasing serum IL-4 levels. CONCLUSION: Liquorice can adjust Th_1/Th_2 imbalance and inhibit the airway inflammation.