Effect of Wumeiwan on Cytokines TNF-α, IL-6, IL-8, IL-10 and Expression of NF-κBp65 in Rats with Ulcerative Colitis

The effects of Wumeiwan （WMW） on TNF-α, IL-6, IL-8, IL-10 and NF-κBp65 in rats with ulcerative colitis （UC） were investigated, the curative effectiveness of WMW vs salicylazosulfapyridine （SASP） was compared, and the action mechanism was analyzed. Fifty-Six Sprague-Dawley （SD） rats were randomly divided into four groups （n=14 in each group, with equal ratio of male and female）： normal control group, model group, SASP group, and WMW group. Except normal control group, the rat UC models in the remaining three groups were established using the method of 2.4-dinitrochlorobenzene （DNCB） immunization and acetic acid local enema. The rats in model group, SASP group, and WMW group were treated with distilled water, SASP, and WMW respectively. The changes in the symptoms and signs were observed, and levels of IL-6, IL-8, TNF-α, IL-10 and the expression of NF-κBp65 in the colonic tissues were statistically analyzed. The results showed that the levels of IL-6, IL-8, and TNF-α were significantly increased （P〈0.01）, while those of IL-10 significantly reduced （P〈0.01） after establishment of rat UC models as compared with normal control group. The levels of IL-6, IL-8, and TNF-α were obviously lower, but the level of IL-10 was obviously higher in WMW and SASP groups than those in model group （P〈0.05）. The levels of IL-6, IL-8, and TNF-α were lower, while the level oflL-10 was higher in WMW group than in SASP group. NF-κBp65 was expressed negatively or weakly in normal colonic tissues. The positive expression rate of NF-κBp65 in WMW group and SASP group was obviously lower than in model group （P〈0.01）, and there was significant difference between WMW group and SASP group （P〈0.05）. It was concluded that rat UC model was established successfully. WMW could up-regulate the expression of IL-10, down-regulate the expression of TNF-α, IL-6, IL-8, and inhibit the NF-κBp65 activity to adjust immune function, indicating WMW had better curative effects on UC in rats.     

Effect of TGF-β1 on the Expression of IL-12, IL-15, IL-18, IL-4 and IL-10 in Heart Transplantation Rejection in Rats

To investigate the effect of TGF-β1 on the expressions of IL-12, IL-15, IL-18, IL-4 and IL-10 in heart transplantation rejection in rats, a model of rat cervical heterotopic heart transplantation was set up and the model rats were randomly divided into three groups： control group, transplant group and TGF-β1 group. The mRNA expression levels of IL-12, IL-15, IL-18, IL-4 and IL-10 were determined by RT-PCR at the 5th day after the transplantation. The mRNA expression levels of IL- 12, IL-15, IL-18 were increased obviously and those of IL-4, IL-10 were significantly decreased in the transplant group as compared with the control group （P〈0.01）. In the TGF-β1 group, the mRNA ex- pression levels of IL- 12, IL- 15, IL- 18 were significantly decreased and those of IL-4, IL- 10 were significantly increased as compared with the transplant group （P〈0.01）. The immunosuppressive effect of TGF-β1 on heart transplantation rejection was related to its inhibition of the expressions of Th1-type cytokines （IL-12, IL-15, IL-18 etc） and its promotion of the expressions of Th2-tpye cyto- kines （IL-4, IL-10）.     

Effect of expression of TGF-beta and TNF-alpha with Qidan granule treatment in rats by bleomycin-induced pulmonary fibrosis

Objective: To evaluate the therapeutic effects and mechanisms of Qidan granule in blemycinA5-induced pulmonary interstitial fibrosis (PIF)in rats. Methods: PIF models were established by blemycinA5-induced in rats. They were treated by Qidan granule and Hydrocortisone respectively. The pathological changes and collagen protein disposition were observed, and the expression of TGF-β, TNF-α proteins were measured by immunohistochemical technique . Results: The pulmonary alveolitis and fibrosis were alleviated remarkably in Qidan granule group compared with those in the model control group and hydrocortisone group (P <0. 01). The expression of TGF-β and TNF-α protein were higher in Qidan granule group than those in normal group , and were significantly less than those in the model control group and in hydrocortisone group (P < 0. 01). Conclusion: Qidan granule would ameliorate the pulmonary alveolitis and fibrosis. TGF-β and TNF-α might play an important role in the development of alveolitis and fibro     

Effect of Coiquhoumia Root on the Expression of Transforming Growth Factor-β in Mesangiai Proliferation Giomerulonephritis Model

Summary： To study the efficacy and the mechanism of Colquhoumia root （ Tripterygium hypoglaucure （Le,vL） Hutch） in the treatment of mesangial proliferation glomerulonephritis （MsPGN）, SD rats were injected with anti-thymoeyte serum （ATS） to make MsPGN model （anti-Thyl model）. The rats were then divided into 3 groups： normal control group, anti-Thyl model group and treatment group. Histopathologieal （HE, PAS）, immunohistoehemieal, RT-PCR technique and computer imaging analysis system were used to evaluate mesangial matrix production, the expression of TGF-β protein and mRNA in the tissues of kidney. Our result showed that proteinuria and the ratio of extraeellular matrix/glomerular capillaries area （ECM/CA） were increased significantly in model group. The expression of both TGF-β protein and mRNA in glomeruli was much higher in model group than in control group （P〈0.01）. After the treatment with Colquhoumia root, proteinuria, ECM/CA and the expression of both TGF-β1 protein and mRNA in glomeruli were significantly decreased in treatment group as compared with those in model group. It is concluded that Colquhoumia root is effective in reducing proteinuria and mesangial matrix proliferation in MsPGN and it may achieve these effects by inhibiting the expressions of TGF-β1 protein and mRNA of mesangial cells.     

Effects of IL-4 on cyclooxygenase-2 and platelet-derived growth factor in the lungs of COPD rats

Objective： To explore the role of interleukin 4（IL-4）, expressions of cyclooxygenase-2 （COX-2） and platelet-derived growth factor （PDGF） in the model of chronic obstructive pulmonary disease （COPD）, in the lungs of rats. Methods： Male Wistar rats were used to build up the model of COPD. Rats were randomly divided into the control group and model group, the IL-4 group and the dexamethasone group. The expressions of COX-2, PDGF-A and PDGF-B in the lung tissue were detected by western blotting and RT-PCR. Results： The expressions of COX-2, PDGF-A and PDGF-B in the model group were increased significantly. Those expressions in the IL-4 and dexamethasone group were notably decreased. Conclusion： IL-4 and dexamethasone could interfere in the establishment of COPD. The expressions of COX-2 and PDGF in the lung tissue of COPD were increased significantly and IL-4 and dexamethasone could decrease those expressions.     

Effect of IL-1O gene transmission on the PTg-stimulated splenocytes proliferation and Thl cytokines production from experimental autoimminue thyroiditis rats

Objective To investigate the effects of gene therapy with IL-10 on PTg-induced proliferation of splenocytes and Th1 cytokine production from PTg-stimulated splenocytes. Methods EAT rats were divided into four groupsgroup A (PBS PLL) , group B (pORF PLL), group C (pORFmIL10 PLL), and group D (pORFmIL10 MEM). The substances mixed with lipofectamine were injected into the thyroid tissues of rats on the 18th dday after immunization. The rats were sacrificed at the 8th week. In vitro proliferative responses to ConA and different concentration of PTg were measured by culturing 4×105 splenocytes pulsed with 18.SKBq of [3H] thymidine for the final 12h and then harvested for liquid scintillation counting. In vitro splenocytes were cultured with PTg (25 mg/L). Th1 cytokine IFN-γ,TNF-αand IL-2 were detected by ELISA. Results The proliferative response to PTg was suppressed in group C, compared with that of group A and B (P＜0.05). The levels of IFN-γ,TNF-oand IL-2 in the supernatant of PTg-stimulated splenocytes were 3548.25 ± 779.47 pg/ml, 27.66±10.50 pg/ml and 3617.73± 609.15 pg/ml, respectively,which were much lower in group C than those in group A and B(P＜0.01, P＜0.05, P＜0.001, respectively). Conclusion IL-10 gene transmission in thyroid tissues could inhibit PTg specific proliferation of splenocytes from EAT rats and the secretion of Thl cytokines from PTg-stimulated splenocytes.     

肠炎清通过抑制NF-κB活性对大鼠肠黏膜起抗炎作用

目的 检测大鼠小肠炎模型肠黏膜通透性的改变,探讨肠炎清对大鼠小肠炎模型肠黏膜通透性的作用的机制.方法 应用氨甲碟呤制备大鼠小肠炎模型,实验设正常对照组、模型对照组、N-乙酰半胱氨酸(NAC,100 mg/kg)组及肠炎清组(CYQ,100mg/kg),每天灌胃给药1次,共6d.每天观察大鼠疾病活动指数(DAI)和肠黏膜损伤指数(CMDI),光镜下观察组织学改变并评分(HS).ELISA测定大鼠IL-10,RT-PCR方法检测大鼠肠黏膜内TNF-α、IL-β的表达,应用Western blotting测定各处理组细胞内磷酸化胞质IκB蛋白表达水平.结果 与正常组相比,小肠炎模型组DAI、CMDI、HS评分明显升高(P<0.01).肠炎清组DAI,CMDI,HS评分较模型组有明显下降(P<0.01).MTX组TNF-α、IL-1βmRNA达水平均较正常对照组明显增高,肠炎清组与NAC治疗组TNF-α、IL-1β表达水平低于MTX组,但IL-10的表达高于MTX组.肠炎清能抑制IκB的降解.结论 姜黄素对肠黏膜起保护作用的机制可能是抑制NF-κB活性,进而减少致炎细胞因子TNF-α、IL-1βmRNA表达,增强抑炎因子IL-10的表达来实现的

Abstract：

Objective To detect the changes in intestinal mucosal permeation in rats with methotrexate-induced small intestinal damage and investigate the protective effects of Changyanqing decoction. Methods Rat enteritis model was established by methotrexate (MTX) and sodium chloride.The rats were randomly divided into normal control group,model group,N-acetylcysteine(NAC) group and Changyanqing decoction group,and Changyanqing decoction(100 mg/kg)or saline was administered daily in the corresponding groups by gastric irrigation for 6 days.The disease activity index(DAI),colonic mucosal damage index(CMDI)and histological score(HS) of the rats were observed and evaluated.The levels of mRNA expressions of TNF-α and IL-1β were detected by semi-quantitative RT-PCR.The expression of IL-10 was detected by enzyme linked immunosorbent assay,and IκB expression was determined with Western blotting.Results Compared with the normal control group,the model group showed significantly increased DAI,CMDI and HS.The DAI,CMDI,and HS in rats treated with Changyanqing decoction were significantly decreased in comparison with those in the model group (P<0.01).The expressions of TNF-α and IL-1β were significantly higher in MTX-treated group than in the control group.The expression of TNF-α and IL-1β mRNA in the Changyanqing group and NAC group were significantly lower,but IL-10 significantly higher than those of the MTX group.In MTX group,obvious NF-κB activation was observed,whose expression was significantly stronger in the cell nuclei,and the IκB in the cytoplasm was markedly degraded.Conclusion Changyanqing decoction offers protection on intestinal mucosa by inhibiting NF-κB activation to reduce TNF-α and IL-1β mRNA expressions and increase IL-10 expression.     

TNF-α and IL-8 of the Patients with Allergic Asthma

The levels of serum TNF-α and IL-8 in the patients with allergic asthma during acute attack period and remission period, and the effects of glucocorticoid (GC) on them were investigated. By using ELISA, the levels of TNF-α and IL-8 were detected in the healthy volunteers (group C, n=40), the patients with allergic asthma (n=40) during acute attack period (group A) and remission period (group B) and those taking GC for a week (n=28). The results were compared among them. It was found that the levels of TNF-α and IL-8 in group A were higher than in group B and group C. In the patients subject to GC therapy, the levels of TNF-α and IL-8 were decreased as compared with those in group A. In group B, the level of TNF-α was higher than in group C, but there was no significant difference in the level of IL-8 between group B and group C. It was concluded that the inflammatory cytokines, TNF-α and 11.-8, played important roles in the bronchus allergic inflammation. GC could reduce the levels of serum TNF-α and IL-8 to exert the anti-inflammatory effects.     

Plasma Levels of the Anti-inflammatory Cytokine IL-10 and Inflammatory Cytokine IL-6 in Patients with Unstable Angina

The plasma levels of inflammatory cytokine interleukin-6 （IL-6） and anti-inflammatory cytokine interleukin-10 （IL-10） in the patients with unstable angina or stable angina were determined and compared. In 30 patients with unstable angina and 22 patients with stable angina, plasma levels of IL-10 and IL-6 were detected by ELISA and plasma lipid parameters by lipid research clinical methods respectively. The results showed plasma levels of IL-10 were significantly lower in unstable angina group than in stable angina group （P=0. 005）, while those of IL-6 were significantly increased in unstable angina group as compared with those in stable angina group （P= 0. 039）. There was a significantly negative correlation between IL-10 and IL-6 in patients with unstable angina （r=-0.41, P=0. 003）. In the unstable angina group, IL-6 levels were obviously positively correlated with TC （r=0. 314, P=0. 023）, but not with TG and HDL. There were no significant correlations between IL-10 and plasma lipid parameters. It was suggested that the decreased IL-10 and increased IL-6 might be associated with the atheromatous plaque stability and progression of coronary heart diseases. IL-10 may play an important role in preventing coronary vascular lesions.     

Effects of angiotensin Ⅱ receptor antagonist on expression of collagen Ⅲ, collagen Ⅴ, and transforming growth factor β1 in the airway walls of sensitized rats

Background Repeated attacks of bronchial asthma lead to different degrees of airway remodeling, the mechanism of which is not yet clear. Some evidences indicate that it is related to the excessive expression of some growth promotion factors. Angiotensin Ⅱ is a polypeptide that may be involved in airway remodeling. To evaluate its role in airway remodeling in asthma, we observed the effects of an angiotensin Ⅱ type 1 receptor antagonist (valsartan) on the expression of collagen Ⅲ, collagen Ⅴ, and transforming growth factor β1 (TGF-β1) mRNA and protein in the airway walls of sensitized rats.Methods Forty Wistar rats were randomly divided into 5 groups: control group, sensitized group, and valsartan groups 1, 2, and 3. The rats in the sensitized group and in valsartan groups 1, 2, and 3 were sensitized and challenged with ovalbumin. Rats in control group were sensitized and challenged with 0.9% NaCl. Rats from valsartan groups 1, 2, and 3 were drenched with valsartan (10 μg, 20 μg, or 30 μg, respectively) at the time of the ovalbumin challenges. The expression of collagen Ⅲ, collagen Ⅴ, and TGF-β1 protein were detected using immunohistochemical method in combination with image analysis methods. The expression of TGF-β1 mRNA was detected by in situ hybridization. Results The expression in the airways of collagen Ⅲ and collagen Ⅴ was significantly higher in rats from the sensitized group (7.73±0.81, 1.34±0.28) and from valsartan groups 1, 2, and 3 (5.73±0.64, 1.13±0.15; 4.96±0.51, 0.98±0.08; 4.43±0.35, 0.93±0.06, respectively) than those in the control group (2.65±0.38, 0.67±0.08, P&lt;0.05). In addition, collagen levels were significantly lower in valsartan groups 1, 2, and 3 than those from the sensitized group (P&lt;0.05). The expression of TGF-β1 mRNA and protein in the airways was significantly higher in rats from the sensitized group (20.49%±3.46%, 29.73%±3.25%) and from valsartan groups 1, 2, and 3 (16.47%±1.94%, 19.41%±1.87%; 14.38%±1.58%, 18.29%±1.43%; 12.96%±1.73%, 18.63%±1.11%, respectively) than that from the control group (7.84%±1.61%, 5.63%±1.07%, P&lt;0.05). TGF-β1 mRNA and protein levels were significantly lower in valsartan groups 1, 2, and 3 than that in the sensitized group (P&lt;0.05). Conclusions Angiotensin Ⅱ receptor antagonist valsartan can suppress synthesis of collagen Ⅲ and collagen Ⅴ by downregulating TGF-β1 mRNA and protein expression. Valsartan can decrease airway remodeling and could play a role in asthma therapy.     

Effects of propofol on early and late cytokines in lipopolysaccharide-induced septic shock in rats

Objective: It has been reported that the intravenous anesthetic propofol (PPF) has anti-inflammatory effects in vitro and in patients. The purpose of this study was to investigate whether PPF has anti-inflammatory effects in lipopolysaccharide (LPS)-induced septic shock by inhibiting the induction of pro-inflammatory cytokines [inter-leukin-6 (IL-6) and tumor necrosis factor-α (TNF-α)] and high mobility group box 1 (HMGB1) in rats. Methods: Thirty six male Wistar rats were randomly assigned to one of three groups (control group, PPF + LPS group and LPS group; n = 12 per group). Control group rats received a 0.9% NaCl solution (NS) by the tail vein. The PPF + LPS group rats received PPF (10 mg/kg bolus, followed by infusion at 10 mg/(kg·h) through a femoral vein cath-eter) 1 h before LPS (7.5 mg/kg) administration via the tail vein. The LPS group rats received injection of LPS (7.5 mg/kg) via the tail vein. Hemodynamic effects were recorded as well as mortality rates, and plasma cytokine con-centrations (TNF-α, IL-6, HMGB1) were measured for the 24-h observation period. Results: The mean arterial pressure and heart rate of the PPF + LPS group were more stable than those of the LPS group. The mortality at 24 h after the administration of the LPS injection was much higher in the LPS group (58.3%) compared to the PPF + LPS group (25.0%). Plasma concentrations of cytokines (IL-6 and TNF-α) and HMGB1 were significantly reduced in the PPF + LPS group compared to the LPS group (P < 0.05). Conclusion: Pretreatment with PPF reduced the mortality rate of rats and attenuated the pro-inflammatory cytokine responses in an endotoxin shock model through an anti-inflammatory action inhibiting induction of HMGB1.     

Effects of propofol on early and late cytokines in lipopolysaccharideinduced septic shock in rats

Objective：It has been reported that the intravenous anesthetic propofol（PPF）has anti-inflammatory effects in vitro and in patients.The purpose of this study was to investigate whether PPF has anti-inflammatory effects in lipopolysaccharide（LPS）-induced septic shock by inhibiting the induction of pro-inflammatory cytokinesinterleukin-6（IL-6）and tumor necrosis factor-α（TNF-α）and high mobility group box 1（HMGB1）in rats.Methods：Thirty six male Wistar rats were randomly assigned to one of three groups（control group,PPF＋LPS group and LPS group;n=12 per group）.Control group rats received a 0.9%NaCl solution（NS）by the tail vein.The PPF＋ LPS group rats received PPF（10 mg/kg bolus,followed by infusion at 10 mg/（kg·h）through a femoral vein catheter）1 h before LPS（7.5 mg/kg）administration via the tail vein.The LPS group rats received injection of LPS（7.5 mg/kg）via the tail vein.Hemodynamic effects were recorded as well as mortality rates,and plasma cytokine concentrations（TNF-α,IL-6,HMGB1）were measured for the 24-h observation period.Results：The mean arterial pressure and heart rate of the PPF＋LPS group were more stable than those of the LPS group.The mortality at 24 h after the administration of the LPS injection was much higher in the LPS group（58.3%）compared to the PPF＋ LPS group（25.0%）.Plasma concentrations of cytokines（IL-6 and TNF-α）and HMGB1 were significantly reduced in the PPF＋LPS group compared to the LPS group（P0.05）.Conclusion：Pretreatment with PPF reduced the mortality rate of rats and attenuated the pro-inflammatory cytokine responses in an endotoxin shock model through an anti-inflammatory action inhibiting induction of HMGB1.     

Impaired upregulation of keratinocyte growth factor in injured lungs induced by Pseudomonas aeruginosa in immunosuppressed rats

Background The number of immunosupressed patients has increased in the past decades. Among them Pseudomonas aeruginosa （P. aeruginosa） is one of the leading bacteria for pneumonia that are associated with poor prognosis. However, the pathogenesis of P. aeruginosa pneumonia in immunosupressed patients is not understood completely. Previous reports showed keratinocyte growth factor （KGF） is associated with lung injury in immunocompetent hosts. In this study, we investigated the different reactions of lung injury, lung pathology and KGF expressions in P aeruginosa pneumonia between immunosuppressed and immunocompetent rats. Methods Immunosuppression of male rats was induced by injecting immunosuppressive subcutaneously. Pneumonia was established by instilling P aeruginous tracheally. The immunocompetent rats were the control group. Survival rate, lung histopathology, pulmonary permeability and oedema, KGF mRNA and protein expressions in lungs of both groups were investigated. Results The survival rate of immunosuppressed group was lower than that of immunocompetent group （33.3% vs 83.3%）. After exposure to bacteria, pulmonary permeability and wet/dry ratio in immunosuppressed group were higher than those in immunocompetent group. Pulmonary congestion and haemorrhage were more intensive in immunosuppressed group compared to immunocompetent group. Apoptosis and necrosis were also observed in infected lungs of immunosuppressed rats. Although we detected KGF expressions in lungs of both groups after infection, the expressions of KGF protein and mRNA gene in immunosuppressed group were much lower than in immunocompetent group. Conclusions Compared with immunocompetent group, there was more intensive lung injury in immunosuppressed group. Severe lung injury may contribute to the poor prognosis of pneumonia. KGF expressions of pneumonia in immunosuppressed rats were less than those in immunocompetent ones.     

Effects of propofol on the mRNA expression of toll-like receptor 4 in BV-2 cells during mimic ischemia-reperfusion injury <Emphasis Type="Italic">in vitro</Emphasis>

This study investigated the effects of propofol on the mRNA expression of Toll-like receptor-4 （TLR4） in BV-2 cells during mimic ischemia-reperfusion （I/R） injury in vitro. BV-2 cells, a mouse microglia line, were cultured and divided into 4 groups at random： control group （group C）, ischemia/reperfusion group （group I/R）, low-dose propofol （25 μmol/L） intervention group （group PF25） and high-dose propofol （100 μmol/L） intervention group （group PF100）. The mRNA expression of TLR4 and NF-κB was measured by means of RT-PCR. TNF-α levels in the supernatants of BV-2 cells were detected by ELISA. The results showed that the mRNA expression of TLR4 and NF-κB was significantly higher in groups I/R, PF25 and PF100 than in group C （P〈0.01）. And the TNF-α level in the supernatants was elevated in groups I/R, PF25 and PF100 as compared with that in group C （P〈0.01）. After pre-treatment with propofol, the mRNA expressions of TLR4 and NF-κB and the TNF-α level were significantly decreased in groups PF25 and PF100 in comparison to those in group I/R （P〈0.01）. And the decrease in those indicators was more significant in group PF100 than in group PF25 （P〈0.01）. It was concluded that propofol exerted brain-protecting effects during I/R injury by suppressing the mRNA expressions of TLR4 and NF-κB and deceasing the TNF-α level.     

Effect of IL-10 gene transmission on the PTg-stimulated splenocytes proliferation and Th1 cytokines production from experimental autoimminue thyroiditis rats

Objective: To investigate the effects of gene therapy with IL-10 on PTg-induced proliferation of splenocytes and Th1 cytokine production from PTg-stimulated splenocytes. Methods: EAT rats were divided into four groups:group A (PBS PLL) , group B (pORF PLL), group C (pORFmIL10 PLL), and group D (pORFmIL10 MEM). The substances mixed with lipofectamine were injected into the thyroid tissues of rats on the 18th dday after immunization. The rats were sacrificed at the 8th week. In vitro proliferative responses to ConA and different concentration of PTg were measured by culturing 4×105 splenocytes pulsed with 18.SKBq of [3H] thymidine for the final 12h and then harvested for liquid scintillation counting. In vitro splenocytes were cultured with PTg (25 mg/L). Th1 cytokine IFN-γ,TNF-αand IL-2 were detected by ELISA. Results: The proliferative response to PTg was suppressed in group C, compared with that of group A and B (P＜0.05). The levels of IFN-γ,TNF-oand IL-2 in the supernatant of PTg-stimulated splenocytes were 3548.25 ± 779.47 pg/ml, 27.66±10.50 pg/ml and 3617.73± 609.15 pg/ml, respectively,which were much lower in group C than those in group A and B(P＜0.01, P＜0.05, P＜0.001, respectively). Conclusion: IL-10 gene transmission in thyroid tissues could inhibit PTg specific proliferation of splenocytes from EAT rats and the secretion of Thl cytokines from PTg-stimulated splenocytes.     

Effects of Propofol on The mRNA Expression of Toll-like Receptor 4 in BV-2 Cells During Mimic Ischemia-Reperfusion Injury In Vitro

This study investigated the effects of propofol on the mRNA expression of Toll-like receptor-4 (TLR4) in BV-2 cells during mimic ischemia-reperfusion (I/R) injury in vitro. BV-2 cells, a mouse microglia line, were cultured and divided into 4 groups at random: control group (group C), ischemia/reperfusion group (group I/R), low-dose propofol (25 μmol/L) intervention group (group PF25) and high-dose propofol (100 μmol/L) intervention group (group PF100). The mRNA expression of TLR4 and NF-κB was measured by means of RT-PCR. TNF-α levels in the supernatants of BV-2 cells were detected by ELISA. The results showed that the mRNA expression of TLR4 and NF-κB was significantly higher in groups I/R, PF25 and PF100 than in group C (P<0.01). And the TNF-α level in the supernatants was elevated in groups I/R, PF25 and PF100 as compared with that in group C (P<0.01). After pre-treatment with propofol, the mRNA expressions of TLR4 and NF-κB and the TNF-α level were significantly decreased in groups PF25 and PF100 in comparison to those in group I/R (P<0.01). And the decrease in those indicators was more significant in group PF100 than in group PF25 (P<0.01). It was concluded that propofol exerted brain-protecting effects during I/R injury by suppressing the mRNA expressions of TLR4 and NF-κB and deceasing the TNF-α level.     

Pioglitazone ameliorates experimental nonalcoholic steatohepatitis by regulating hepatic nuclear factor-kappa B and cyclooxygenases-2 expression in rats

Background Pioglitazone is effective in nonalcoholic steatohepatitis (NASH), but the mechanism of action is not completely understood. This study was designed to investigate the impact of pioglitazone on hepatic nuclear factor-kappa B (NF-κB) and cyclooxygenases-2 (COX-2) expressions in NASH rats. Methods Thirty Sprague-Dawley male rats were randomly assigned to a control group (n=10), NASH group (n=10) and pioglitazone treatment group (n=10). Liver tissues were processed for histology by HE and Masson stained. Biochemical parameters of antioxidant enzyme activities, tumor necrosis factor alpha (TNF-α), prostaglandin E2 (PGE2) in the serum and hepatic samples were measured. The mRNA and protein expressions of peroxisome proliferator-activated receptor gamma (PPARγ), NF-κB and COX-2 were determined by real-time polymerase chain reaction, Western blotting and immunohistochemistry, respectively. Results There were severe steatosis, moderate inflammatory cellular infiltration and fibrosis in the livers of the NASH models. After treatment, steatosis, inflammation and fibrosis were significantly improved compared with the NASH group (χ2＝20.40, P＜0.001; χ2＝20.17, P＜0.001; χ2＝13.98, P=0.002). The serum and hepatic levels of total anti-oxidation competence (T-AOC), superoxide dismutase (SOD), catalase(CAT), glutathione peroxidase (GSH-PX) and malondialdehyde (MDA) in the NASH group were conspicuous disordered than those indicators in the control group. In the NASH group, the levels of serum TNF-α and PGE2 were significantly increased compared with the control group. Immunohistochemistry showed expressions of NF-κB and COX-2 in livers were significantly elevated, but PPARγ was decreased in the NASH group. Real-time PCR and Western blotting revealed mRNA and protein expressions of COX-2 were increased in the NASH group compared with the control group (0.57±0.08 vs 2.83±0.24; 0.38±0.03 vs 1.00±0.03, P＜0.001 and P=0.004, respectively). After pioglitazone intervention, all of those indicators markedly improved(P＜0.05 or P＜0.01 ). Conclusion Suppressing hepatic NF-κB and COX-2 expression, at least in part, is one of the possible therapeutic mechanisms of pioglitazone in NASH rats.     

Intrathecal co-administration of ketamine and morphine preventing activation of astrocytes and decreasing releases of IL-1β and IL-6 from spinal cord in rats of morphine tolerance

Objective: To investigate the effects of intrathecal administration of ketamine, a non-competitive N-methy-D-aspartate receptor antagonist, combined with morphine on the activation of astrocytes and releases of IL-1β and IL-6 from spinal cord in the rats of morphine tolerance. Methods: Twenty-seven Sprague-Dawley male rats were randomly divided into sham-operated, morphine tolerance, and morphine plus ketamine group. The subarachnoid catheterization of all the rats was prepared by the method of Jianping Yang.Morphine 20 μg in 10 μl was administrated intrathecally to induce spinal morphine tolerance once daily for 5 consecutive days. Morphine and ketamine 250 μg in 10 μl total volume was given in morphine plus ketamine group. Three groups all received intrathecal morphine 5 μg in 10 μl for morphine challenge test at 24 h after last administration of the morphine. After morphine challenge test, lumbar spinal tissues were taken for measurement of glial fibrillary acidic protein (GFAP) of astrocyte in lumbar spinal horn cord by immunohistochemistry and IL-1βand IL-6 of spinal cord by ELISA. Results: The decrease of ％MPE induced by chronic intrathecal morphine was inhibited by ketamine and hyperalgesia and allodynia induced by morphine-withdrawl were alleviated. The average areas, the average absorbency (-A), the integral absorbency (A) of GFAP immuno-reactive cells in the dorsal horn, and IL-1β and IL-6 of spinal cord were significantly larger in morphine tolerance group than in morphine plus ketamine group. Conclusion: Co-administration of ketamine and morphine enhance antinociceptive effect of morphine and prevent the development of morphine tolerance. Ketamine might attenuate the activation of astrocytes and inhibit the release of IL-1β and IL-6 from spinal cord in repeated intrathecal morphine rats.