Role of Nuclear Transcription Factor-κB in Endotoxin-induced Shock in Rats

To investigate the role of NF-κB in endotoxic shock in rats. the model of endotoxinshock rats was induced by intravenous infusion of lipopolysaccharidc (LPS). 1 h. 2 h. 4 h and 6 h after LPS injection, the activation of NF-κB in blood mononuclear cells and the content of TNF-α and IL-6 in plasma was detected by enzyme-linked immunoadsordent assay (ELISA). The level of mean arterial pressure (MAP) and the histopathological changes of lung and liver were also observed. The activation of NF-κB in mononuclear cells increased 1 h after LPS injection and reached its peak 2 h after the injection, and its level was higher than that of normal group. The level of TNF-α was increased 1 h after the infusion and peaked 2 h after the injection, and its level was higher than that of normal group after LPS infusion. The content of IL-6 increased gradually with time. the IL-6 level was higher than that of normal group after LPS injection. MAP was decreased gradually with time and its level was lower than that of normal group after LPS injection. Pathological examination showed that endotoxic shock could cause pulmonary alveolar hemorrhage, edema and infiltration of inflammatory cell in lung tissue and congestion, edema, capillary dilation and inflammatory cell infiltration in liver tissue. It is concluded that NF-κB can up-regulate the expression of TNF-α and IL-6 in plasma and play an important role in endotoxin induced shock in rats.     

Protective Effect of Hydroxysafflor Yellow A on Inflammatory Injury in Chronic Obstructive Pulmonary Disease Rats

Objective: To investigate the attenuating effect of Hydroxysafflor yellow A(HSYA) on inflammatory injury in chronic obstructive pulmonary disease(COPD). Methods: Rats were randomly assigned to 7 groups according to body weight including normal control group, HSYA blank group(76.8 mg/kg), COPD group, COPD+HSYA(30, 48, 76.8 mg/kg) groups and COPD+dexamethasone(2 mg/kg), 10 in each group. Passive cigarette smoke and intratracheal instil ation of lipopolysaccharides were used to establish a COPD model in rats. Hematoxylin and eosin staining of lung tissue sections was used, real-time polymerase chain reaction(PCR) was used to assay m RNA levels of some cytokines in lung tissues, the cytokines in bronchoalveolar lavage fluid(BALF) were measured by enzyme-linked immunosorbent assay(ELISA), Western blot analysis was used to determine phosphorylated p38 mitogen-activated protein kinase(MAPK) levels in lung tissues, and nuclear factor-κB(NF-κB) p65 protein levels in lung tissues were detected by immunohistochemistry. Results: Lung alveolar septa destruction, alveolus fusion, inflammatory cel infiltration, and bronchiole exudation were observed. These pathological changes were al eviated in the COPD+HSYA group. The m RNA expression of inflammatory factors were significantly increased in lung tissues from COPD rats(all P0.01) and were inhibited by HSYA. Levels of inflammatory cytokines in BALF of COPD rats were significantly increased(all P0.01) which were inhibited by HSYA(all P0.01, 48, 76.8 mg/kg). The levels of p38 MAPK phosphorylation and p65 in lung tissues of COPD rats were significantly increased(al P0.01) and were suppressed by HSYA(all P0.01, 48, 76.8 mg/kg). Conclusions: HSYA could alleviate inflammatory cell infiltration and other pathological changes in the lungs of COPD rats. HSYA could inhibit inflammatory cytokine expression, and increase phosphorylation of p38 MAPK and NF-κB p65 in the lungs of COPD rats. The protective mechanism of HSYA to inhibit COPD inflammation might be by attenuating NF-κB and p38 MAPK signal transduction.     

GMCSF对高氧暴露新生大鼠肺组织RAGE-NF-κB通路的影响

目的 探讨高氧暴露对新生大鼠肺组织高级糖基化终产物受体(RAGE)-核因子κB(NF-κB)信号通路的影响及粒细胞巨噬细胞集落刺激因子(GMCSF)肺损伤保护作用的相关机制.方法 24只3日龄新生大鼠随机平均分为3组:正常对照组(空气环境下7 d)、高氧对照组(95%氧暴露7 d)、高氧干预组(95%氧暴露7 d+GMCSF皮下注射9 μg/kg/次,共3次);光镜观察并盲法进行肺组织病理学损伤评分;RT-PCR法检测肺组织匀浆RAGE mRNA、NF-κB mRNA表达;Western印迹检测肺组织匀浆RAGE、NF-κB蛋白表达;ELISA检测支气管肺泡灌洗液(BALF)及血清中肿瘤坏死因子-α(TNF-α)水平;免疫组化法检测肺组织石蜡切片RAGE表达情况.结果 正常对照组、高氧对照组、高氧干预组肺组织损伤评分分别为0.46±0.20、3.06±0.33、2.31±0.56,差异有统计学意义(P=0.000);3组RAGE mRNA和蛋白表达分别为0.14±0.02、0.34±0.06、0.28±0.04和0.30±0.04、0.76±0.11、0.55±0.08,差异均有统计学意义(均P=0.000);3组NF-κB mRNA和蛋白表达分别为0.41±0.21、0.90±0.36、0.69±0.30和0.41±0.26、0.96±0.43、0.77±0.33,差异均有统计学意义(P=0.000和P=0.017);3组BALF中TNF-α水平分别为76±10、224±42、143±24,差异有统计学意义(P=0.000).以上各指标结果在高氧对照组、高氧干预组均明显高于正常对照组(均P<0.05),高氧干预组低于高氧对照组(P<0.05).3组血清TNF-α水平差异无统计学意义(P>0.05).结论 GMCSF可能通过下调RAGE-NF-κB信号通路对高氧肺损伤发挥保护作用.

Abstract：

Objective To explore the effects of granulocyte-macrophage colony-stimulating factor (GMCSF) on hyperoxia exposure lung injury in newborn rats and elucidate its protective mechanism of operating via the signaling pathway of advanced glycation end products (RAGE)-NF-κB.Methods Twenty-four 3-day-old SD rats from 3 litters were randomly divided into 3 groups. They were hyperoxia exposure plus GMCSF group (group A), hyperoxia exposure group (group B) and air exposure group (group C). The rats from groups A and B were placed in a sealed Plexiglas chamber with a minimal in-and-outflow, providing 6-7 exchanges per hour of chamber volume and maintaining O2 levels above 95%.While the rats in group C only were exposed to air simultaneously.The rats in group A received subcutaneous injections of recombinant murine GMCSF (9 μg/kg) during hyperoxia exposure at 24 h, 72 h and 120 h respectively. And the rats in groups B and C received subcutaneous injections of saline vehicle alone at the same time point. Seven days later, all were sacrificed and immunohistochemistry was employed to assess the expression of RAGE in lung tissue. The levels of tumor necrosis factor-α in bronchoalveolar lavage fluid (BALF) and serum samples were detected by ELISA (enzyme-linked immunosorbent assay). The RAGE mRNA and NF-κB mRNA in tissue homogenates were detected by RT-PCR while RAGE and NF-κB by Western blot. Also the values of lung damage score were calculated with microscopic histology.Results The value of lung damage score in group C, B and A was 0.46±0.20, 3.06±0.33 and 2.31±0.56 respectively, there was significantly difference among three groups (P=0.000). The expression of RAGE mRNA and protein in three groups were 0.14±0.02, 0.34±0.06, 0.28±0.04 and 0.30±0.04, 0.76±0.11, 0.55±0.08 respectively. There were both significantly differences among three groups (P=0.000, P=0.000). The expression of NF-κB mRNA and protein in three groups were 0.41±0.21, 0.90±0.36, 0.69±0.30 and 0.41±0.26, 0.96±0.43, 0.77±0.33 respectively, there were both significantly difference among three groups (P=0.000, P=0.017). The level of TNF-α in BALF was 76±10, 224±42 and 143±24 respectively, there was significantly difference among three groups (P=0.000). All indicators above in group B and group A were significantly more than those in group C (all P<0.05), while these indicators in group A were lower than those in group B. But there was no difference in the level of TNF-α of serum among three groups (P>0.05).Conclusion GMCSF may protect hyperoxia-induced lung injury via down-regulating the signaling pathway of RAGE-NF-κB.     

Coronary microembolization induced myocardial contractile dysfunction and tumor necrosis factor-α mRNA expression partly inhibited by SB203580 through a p38 mitogen-activated protein kinase pathway

Background  The microemboli produced during spontaneous plaque rupture and ulceration and during coronary intervention will reduce coronary reserve and cause cardiac dysfunction. It is though that inflammation caused by the microinfarction induced by the microembolization may play an essential role. It is known that the activation of p38 mitogen-activated protein kinases (MAPK) in both infected and non-infected inflammation in myocardium may cause a contractile dysfunction. But the relation between the activation of p38 MAPK and microembolization is still unknown.

Methods  Sprague-Dawley rats were randomly divided into three groups: Sham group, coronary microembolization (CME) group and SB203580 group (n=10 per group). CME rats were produced by injection of 42 μm microspheres into the left ventricle with occlusion of the ascending aorta. SB203580, a p38 MAPK inhibitor, was injected into the femoral vein after the injection of microspheres to make the SB203580 group. Left ventricular ejection fraction (LVEF) was determined by echocardiography. The protein concentration of P38 MAPK in the myocardium was assessed by Western blotting. The relative expression of mRNA for tumor necrosis factor (TNF)-α was assessed by the technique of semi-quantitative polymerase chain reaction amplification.  

Results  LVEF was depressed at three hours up to 12 hours in the CME group. Increased p38 MAPK activity and TNF-α mRNA expression were observed in the CME group. The administration of SB203580 partly inhibited p38 MAPK activity, but did not fully depress the TNF-α expression, and partly preserved cardiac contractile function.

Conclusions  p38 MAPK is significantly activated by CME and the inhibition of p38 MAPK can partly depress the TNF-α expression and preserve cardiac contractile function.

     

Effect of Dachengqi Decoction on NF-κB p65 Expression in Lung of Rats with Partial Intestinal Obstruction and the Underlying Mechanism

To investigate the effect of Dachengqi decoction on NF-κB p65 expression in lung of rats with partial intestinal obstruction and the underlying mechanism, 30 SD rats were randomly divided into three groups： sham-operation group, model group and Dachengqi decoction treatment group （Dachengqi group）, with 10 animals in each group. The models were made by partially ligating their large intestines outside the body. The pathological changes were analyzed by HE staining. The expression of NF-κB p65 in rats lung were measured by using real-time polymerase chain reaction and immunohistochemistry respectively. Moreover, the expression of caveolin-1 in rats lung was also measured to. Increased edema, interstitial thickening, hemorrhage, and infiltration of inflammatory cells were found in the model group. In contrast, this change was significantly reduced in Dachengqi group as compared with model group. In addition, the up-regulated caveolin-1 and NF-κB p65 were also suppressed by Dachengqi decoction in lung of rats with partial intestinal obstruction. We are led to concluded that the caveolin-l-NF-κB pathway plays an important role in the development of lung injury of rats with partial intestinal obstruction and Dachengqi decoction could down-regulate the expression of caveolin-1 and NF-κB p65 in lung of rats with partial intestinal obstruction.     

Cyclovirobuxinum D Alleviates Cardiac Hypertrophy in Hyperthyroid Rats by Preventing Apoptosis of Cardiac Cells and Inhibiting the p38 Mitogen-Activated Protein Kinase Signaling Pathway

Objective: To investigate the underlying mechanisms of cyclovirobuxinum D(Cvb-D) on alleviating cardiac hypertrophy in rats. Methods: Sprague-Dawley rats were randomly divided into 5 groups: control group; levothyroxine-induced cardiac hypertrophy group(model); levothyroxine-induced cardiac hypertrophy + Cvb-D group(Cvb-D); levothyroxine-induced cardiac hypertrophy + captopril group(captopril); levothyroxine-induced cardiac hypertrophy + SB203580 group(SB203580), n=10 for each group. Rats were daily administered the respective drugs continuously for14 days by gastric gavage. A rat model of cardiac hypertrophy was established by intraperitoneal injection of levothyroxine to investigate whether Cvb-D protects against cardiac hypertrophy by inhibiting the p38 mitogen-activated protein kinase(MAPK) signaling pathway and preventing apoptosis of cardiac cells. Results: Treatment with Cvb-D significantly deceased left ventricle hypertrophy, improved the histopathology, hemodynamic conditions, and cardiac function in rats with cardiac hypertrophy. Compared with the normal control group, in rats with cardiac hypertrophy, expression of bax in the heart and phospho-p38 MAPK protein levels were significantly up-regulated(P0.01 or 0.05), whereas the bcl-2 protein level was downregulated(P0.01). In contrast, Cvb-D treatment reversed the changes in bax and phospho-p38 MAPK protein levels but increased the bcl-2 protein level(P0.01 or 0.05), and these effects were similar to those of captopril and SB203580(a specific p38 MAPK inhibitor) treatment. Furthermore, both Cvb-D, captopril and SB203580 reduced m RNA expression of p38α, p38β, c-fos, and c-jun m RNA, and Cvb-D had a stronger effect(P0.01). Conclusion: These results demonstrate that Cvb-D protects against cardiac hypertrophy, which is possibly mediated by prevention of cardiac cell apoptosis and inhibition of the p38 MAPK signaling pathway.     

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.     

Alteration of NF-kB and TNF-α mRNA and protein in hippocampus in the chronic constrictive injury model of rats

Objective To investigate the alteration of nuclear factor kappa B( NF-κB) and tumor necrosis factor-a (TNF-α) mRNA and protein in hippocampus in chronic constrictive injury (CCI) model of rats. Methods Seventy-six male Wistar rats were randomly divided into 2 groups ( n = 38): the CCI group which received the chronic constriction injury and the sham group which received the sham operation as control. The mechanical and thermal nociceptive thresholds were assessed with paw withdrawal latency (PWL) to von Frey filaments and radiant heat at 1d before and ld,4d,7d,14d and 28d after CCI operation. Five animals were sacrificed at each time point for real-time polymerase chain reaction (real-time PCR) and another three animals sacrificed at 7d postoperation for immunofluorescence histochemical staining. Results The thresholds to mechanical and thermal stimuli decreased obviously after operation in CCI group. The expressions of TNF-α and NF-κB mRNA began to increase at ld( (2.079 ±0. 104)times and 4d( ( 1.640 ± 0.064) times) after operation and reached the peak at 7d ((2.748 ±0.147)times, (2.010 ±0.096)times) ,then the expressions of TNF-a mRNA began to decrease,while the expressions of NF-kB mRNA maintained at a high level throughout the experiment. The result of immunofluorescence histochemical staining revealed that NF-kB and TNF-α protein expressions at 7 day increased significantly on the hippocampus,which was consisted with NF-κB and TNF-a mRNA levels. Conclusion The activation NF-κB and TNF-α in hippocampus may be involved in the procession of neuropathic pain.     

神经病理性疼痛大鼠海马NF-κB及TNF-α表达水平的变化

目的 观察慢性坐骨神经挤压性损伤(CCI)模型大鼠海马核因子-κB(NF-κB)及肿瘤坏死因子-α(TNF-α)表达的变化规律.方法 雄性Wistar大鼠76只,随机分为手术组(CCI组)和假手术组(Sham组)(每组n=38),于CCI术前1天,术后1,4,7,14,21,28d各时间点测定机械痛阈及热痛阈后立即处死大鼠,取海马组织,采用荧光实时定量PCR方法测定NF-κB及TNF-α mRNA含量的变化,并与术后第7天采用免疫荧光的方法测定两种因子蛋白的表达.结果 与假手术组相比,CCI组大鼠手术侧机械痛阈及热痛阈明显降低.TNF-α及NF-κBmRNA表达水平于CCI术后1d和4 d开始增加,为术前的(2.079±0.104)倍,(1.640±0.064)倍,7 d达到高峰,为术前的(2.748±0.147)倍,(2.010 ± 0.096)倍,随后TNF-αmRNA表达水平迅速下降,而NF-κB mRNA于CCI后28 d仍为术前的(1.439±0.121)倍,维持于较高水平(P＜0.05).术后第7天免疫荧光结果显示NF-κB及TNF-α在海马表达明显增加.结论 CCI致大鼠海马上调NF-κB及TNF-α的表达,可能参与神经病理性疼痛的调控过程.

Abstract：

Objective To investigate the alteration of nuclear factor kappa B( NF-κB) and tumor necrosis factor-a (TNF-α) mRNA and protein in hippocampus in chronic constrictive injury (CCI) model of rats. Methods Seventy-six male Wistar rats were randomly divided into 2 groups ( n = 38): the CCI group which received the chronic constriction injury and the sham group which received the sham operation as control. The mechanical and thermal nociceptive thresholds were assessed with paw withdrawal latency (PWL) to von Frey filaments and radiant heat at 1d before and ld,4d,7d,14d and 28d after CCI operation. Five animals were sacrificed at each time point for real-time polymerase chain reaction (real-time PCR) and another three animals sacrificed at 7d postoperation for immunofluorescence histochemical staining. Results The thresholds to mechanical and thermal stimuli decreased obviously after operation in CCI group. The expressions of TNF-α and NF-κB mRNA began to increase at ld( (2.079 ±0. 104)times and 4d( ( 1.640 ± 0.064) times) after operation and reached the peak at 7d ((2.748 ±0.147)times, (2.010 ±0.096)times) ,then the expressions of TNF-a mRNA began to decrease,while the expressions of NF-kB mRNA maintained at a high level throughout the experiment. The result of immunofluorescence histochemical staining revealed that NF-kB and TNF-α protein expressions at 7 day increased significantly on the hippocampus,which was consisted with NF-κB and TNF-a mRNA levels. Conclusion The activation NF-κB and TNF-α in hippocampus may be involved in the procession of neuropathic pain.     

丹红注射液对脂多糖诱导THP-1巨噬细胞促炎因子分泌的影响及机制

目的观察丹红注射液对脂多糖（LPS）诱导的THP-1巨噬细胞促炎因子分泌的影响,并探讨其机制。方法160nmol／L佛波酯孵育THP-1巨噬细胞24h后分为对照组、脂多糖组、丹红组和脂多糖＋丹红组,酶联免疫吸附法检测培养液中促炎因子的含量,Western blot检测核因子κB（NF—κB）、p65和rroll样受体4（TLR4）的蛋白表达水平。结果丹红注射液明显抑制LPS诱导的巨噬细胞肿瘤坏死因子α（TNF-α）、白细胞介素6（IL-6）和白细胞介素1β（IL-1β）释放,下调核因子NF—κB和TLR4的表达。结论丹红注射液抑制脂多糖诱导的炎症反应,可能与其抑制TLR4和NF—κB的表达有关。     

热射病小鼠大脑皮层组织中炎症细胞因子水平及 P38MAPK/P65NF-κB信号通路变化

目的：探讨热射病小鼠大脑皮层组织中白细胞介素1β（IL-1β）、白细胞介素-6（IL-6）、肿瘤坏死因子α（TNF-α）水平和P38丝裂原活化蛋白激酶（P38 MAPK）/P65核转录因子κB （P65NF-κB）信号通路的变化,并阐明其机制。方法：将60只小鼠根据随机数字法分为对照组及热射病1、6和24 h组（热射病模型小鼠出舱后1、6和24 h）,每组15只。观察各组小鼠体质量丢失和肛温,HE染色检测各组小鼠大脑皮层组织的形态表现,ELISA法测定各组小鼠大脑皮层组织中IL-1β、IL-6和TNF-α水平,Western blotting法检测各组小鼠大脑皮层组织中P38MAPK、P65NF-κB、p-P38MAPK和p-P65NF-κB蛋白表达水平。结果：与对照组比较,热射病组小鼠体质量丢失明显增加（P<0.05）,热射病1和6 h组小鼠肛温明显降低（P<0.05）。HE染色,对照组小鼠大脑皮层脑组织形态表现正常;热射病1 h组小鼠大量脑细胞核固缩、核染色深,血管被压缩变形、管壁外大量水肿;热射病6 h组小鼠核固缩细胞数量减少,血管外水肿减轻;热射病24 h组小鼠核固缩脑细胞和血管外水肿少见。与对照组比较,热射病1和6 h组小鼠大脑皮层组织中IL-1β、IL-6、TNF-α水平和p-P38MAPK、p-P65NF-κB蛋白表达水平明显升高（P<0.05）;热射病6 h组小鼠大脑皮层组织中IL-1β、IL-6、TNF-α水平和p-P38MAPK、p-P65NF-κB蛋白表达水平明显低于热射病1 h组（P<0.05）。结论：热射病小鼠大脑皮层组织中炎症细胞因子水平升高,P38MAPK/P65NF-κB信号通路激活,这种中枢神经系统炎症反应可能与其信号通路激活有关。     

牛磺酸对重症急性胰腺炎大鼠枯否细胞p38MAPK信号转导通路的影响

目的 探讨牛磺酸对重症急性胰腺炎(severe acute pancreatitis,SAP)大鼠枯否细胞(KCs)P38MAPK的影响以及对分泌促炎细胞因子TNF-α和IL-1β的作用.方法 48只SD大鼠采用完全随机法分为假手术对照(SO)组、SAP组、SAP+Taut(牛磺酸)组.SAP模型通过胰胆管逆行注射5%牛磺胆酸钠诱导,造模前SAP+Taur组分别于24、12 h从尾静脉内注入牛磺酸(3 mmol/L,0.1 ml/100 g).假手术或造模后分别于12、24 h处死动物,检测其血清中AST、ALT、AMS含量;分离KCs,采用Western blot法检测P38MAPK活化情况,凝胶电泳迁移率法(EMSA)检测KCs中NF-κB DNA的表达,并用ELISA法检测KCs培养上清液中TNF-α和IL-1β蛋白含量.结果 SAP组大鼠血清中AST、ALT、AMS含量均较SO组明显升高(P<0.01);而SAP+Taur组血清中AST、ALT、AMS含量与SAP组比较,明显降低(P<0.05).SAP组各时间点大鼠KCs中p38MAPK活性显著高于SO组(P<0.01),而且NF-κB的表达也明显高于SO组(P<0.01),KCs培养上清液中TNF-α和IL-1β蛋白含量明显高于SO组(P<0.01),但SAP+Taut组大鼠KCs上述指标均显著低于SAP组.结论 牛磺酸可以抑制急性胰腺炎时KCs中p38MAPK的活化,减少NF-κB的表达,从而对促炎细胞因子TNF-α和IL-1β的分泌起抑制作用,可能具有临床应用前景.     

癫持续状态大鼠海马组织中肿瘤坏死因子α和核因子κB的变化及其相关性

目的探讨癫持续状态（SE）大鼠海马组织中肿瘤坏死因子α（TNF-α）和核因子κB（NF-κB）的表达。方法对Wistar大鼠用10%匹罗卡品300 mg/kg腹腔注射（SE组,n=20）,产生SE后60 min终止发作;另设注射等量生理盐水的对照组（n=6）。饲养24 h后取大鼠海马组织,苏木精-伊红染色观察海马CA3区神经元数量,Western blotting测定海马组织NF-κB表达,双抗体夹心酶联免疫法（ELISA）测定海马组织TNF-α表达。采用Pearson相关性分析法分析TNF-α和NF-κB表达的相关性。结果 SE组海马CA3区神经元数量（53.26±2.67）明显小于对照组（103.22±0.83）,差异具有统计学意义（P〈0.001）。SE组大鼠海马组织TNF-α和NF-κB表达水平均显著高于对照组（P〈0.001和P〈0.05）,且两者表达水平呈正相关（r=0.457,P=0.02）。结论 SE后24 h,TNF-α和NF-κB在大鼠海马组织中表达活性增强,且两者表达水平呈正相关。     

肺气虚模型大鼠介导TNF-α-MAPK信号途径对肾AQP2表达的影响

目的:通过观测肺气虚大鼠血浆中肿瘤坏死因子(TNF-α)含量,肾组织水通道蛋白2(AQP2),丝裂原激活蛋白激酶(p38MAPK)和核因子-κB(NF-κBp65)表达的变化,探讨中医藏象中肺与肾在水液代谢之间关系的现代科学内涵。方法:将SPF级健康雄性Wistar大鼠20只随机分为正常组和模型组,采用ELISA法检测血浆TNF-α含量、Western法检测肾组织AQP2、免疫组化法检测肾组织p38MAPK和NF-κB p65表达变化。结果:与正常组比较,模型组血浆中TNF-α含量升高,肾组织AQP2、p38MAPK和NF-κB p65表达上调(P<0.01)。结论:肺气虚状态下肾组织AQP2表达变化可能是通过TNF-α-MAPK信号转导途径实现的。     

P38丝裂原活化蛋白激酶抑制剂对大鼠重症急性胰腺炎的作用

目的 探讨P38丝裂原活化蛋白激酶（P38MAPK）抑制剂（SB203580）对重症急性胰腺炎的治疗作用.方法 将Wistar大鼠63只随机分为三组：假手术组（S组）,重症急性胰腺炎组（P组）,SB203580处理组（T组）,每组21只.SAP模型由5%牛磺胆酸钠大鼠胆胰管逆行注射诱发而成.模型成功后,P组不做处理 T组立即行SB203580 10 mg/kg腹腔注射 S组仅开腹,不做其他处理 术后各组于1 h、2 h、4 h 3个时间点处死大鼠,各时间点酶联免疫吸附测定（ELISA）法测血清TNF-α水平,并取胰腺组织行病理检查,免疫组化法检测p-P38MAPK表达,TUNEL法检测胰腺腺泡细胞凋亡.结果 血清TNF-α水平随SAP病情的进展而升高（P＜0.05）,其水平P组及T组各时间点均较S组高,但TNF-α水平T组各时间点显著低于P组（P＜0.05） p-P38MAPK表达随SAP病情进展而升高（P＜0.05）,1、2、4 h三个时间点P组及T组均升高（P＜0.05）,但T组活性显著低于P组（P＜0.05）：TUNEL显示假手术组胰腺组织存在极少量的凋亡细胞,T、P组细胞凋亡率均高于S组,T组凋亡率高于P组（P＜0.05） 胰腺病理损害光镜下T组明显轻于P组.结论 P38MAPK可能参与大鼠重症急性胰腺炎发病过程,SB203580可能通过抑制P38MAPK活化减少炎症递质释放,增加胰腺腺泡细胞凋亡,减轻SAP病理损害.     

银杏叶提取物对帕金森病模型大鼠脑内神经炎性反应的影响

目的 观察银杏叶提取物对帕金森病（Parkinson's disease,PD）模型大鼠中脑和纹状体神经炎性反应因子及其调控因子核因子-κB （nuclear factor-kappa B,NF-κB）的影响。方法 将32只3月龄雄性SD大鼠采用数字表法随机分为对照组、模型组、低剂量组和高剂量组,每组8只。背部皮下注射鱼藤酮制备PD大鼠模型,低剂量组和高剂量组同时给予银杏叶提取物灌胃共30 d。酶联免疫法检测大鼠脑内纹状体中神经炎性反应因子-肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）及白细胞介素-1β（interleukin-1β,IL-1β）含量变化,同时Western blotting检测大鼠脑内星形胶质细胞、小胶质细胞和NF-κB的表达改变。结果 与对照组相比,模型组大鼠纹状体中TNF-α和IL-1β均明显增加（P<0.01）,大鼠脑内GFAP、Iba1和NF-κB表达明显增加（P<0.05）。银杏叶提取物治疗后明显逆转了上述神经炎性反应,与模型组相比,治疗组GFAP、Iba1和NF-κB蛋白表达明显降低（P<0.05）,大鼠纹状体中TNF-α和IL-1β明显减少（P<0.05）。同时上述改变有明显剂量依赖性。结论 银杏叶提取物能改善PD大鼠脑内的慢性神经炎性反应,保护脑内黑质多巴胺能神经元,为PD的临床治疗提供新的靶点。     

P38信号通路调控帕金森病小鼠黑质NF-κB和iNOS的表达

目的探讨P38丝裂原活化蛋白激酶（P38 MAPK）信号通路与核因子NF-κB、一氧化氮合酶（inducible nitricoxide, iNOS）
之间的关系,通过给予P38 MAPK特异性抑制剂SB203580,研究P38 MAPK信号通路在1-甲基-4-苯基-1,2,3,6-四氢吡啶
（MPTP）制备的帕金森病（PD）小鼠模型的作用机制。方法将小鼠随机分为模型组、干预组和对照组。观察小鼠行为学变化,
采用免疫组织化学和免疫蛋白印迹法观察小鼠黑质酪氨酸羟化酶（TH）、磷酸化P38（p-P38）、NF-κB和iNOS的表达变化;以及
给予SB203580后对上述变化的影响。结果与对照组相比,模型组小鼠出现典型PD症状,TH阳性神经元明显丢失、蛋白水平
下降,P38 MAPK信号通路被激活,其磷酸化产物p-P38表达明显增多,NF-κB和iNOS表达也明显增加;经SB203580干预处理
后,上述变化均减轻。结论P38 MAPK信号通路可能参与激活NF-κB、iNOS而诱导PD模型小鼠黑质多巴胺能神经元的损伤,
采用SB203580抑制P38 MAPK信号通路可对PD模型小鼠多巴胺神经元起到保护作用。
     

p38丝裂原活化蛋白激酶在溃疡性结肠炎中的作用

目的 探讨p38丝裂原活化蛋白激酶(p38MAPK)在溃疡性结肠炎(UC)以及葡聚糖硫酸钠(DSS)诱导的小鼠结肠炎中的作用。 方法 ①7只Balb/c小鼠随机分为3组:正常对照组、DSS结肠炎组、p38MAPK选择性抑制剂SB203580干预组。结肠炎组小鼠每日饮用5% DSS溶液,干预组小鼠则在饮用5% DSS溶液72 h后,加用SB203580[1 mg/(kg·d)]进行腹腔注射。观察并记录各组小鼠的疾病活动指数(DAI);7 d后处死小鼠,观察各组小鼠肠黏膜组织中肿瘤坏死因子α(TNF-α)和白细胞介素-1β(IL-1β)的表达。②收集36例活动性UC患者的肠黏膜组织标本,并以18例结肠癌旁的正常组织标本作对照,通过体外组织培养,观察SB203580对UC患者肠黏膜活检组织中TNF-α和IL-1β表达的影响。 结果 ①DSS结肠炎小鼠的DAI评分、肠黏膜组织中TNF-α和IL-1β的水平明显升高(P均<0.01),经SB203580干预后,其DAI评分、TNF-α、IL-1β的水平均明显降低,但仍然高于正常对照组(P均<0.01)。②体外组织培养结果显示,与未用SB203580处理的UC组比较,SB203580处理后UC患者肠黏膜组织中TNF-α和IL-1β的水平明显降低(P<0.01)。 结论 SB203580能阻断p38MAPK信号转导通路,从而降低促炎性细胞因子TNF-α和IL-1β的释放。     

阿托伐他汀对脑出血大鼠脑组织NF-κB、TNF-α和IL-1β表达的影响

目的 观察阿托伐他汀对脑出血大鼠血肿周围组织中NF-κB、TNF-α和IL-1β表达的影响,探讨阿托伐他汀对脑出血大鼠的脑保护作用及可能的机制.方法 将120只SD大鼠按随机数字表法随机分为假手术组、脑出血组和治疗组.脑出血组和治疗组采用自体血立体定向注射法,建立脑出血动物模型.假手术组于相同部位注射等量0.9%氯化钠注射液.治疗组于术后2 h将阿托伐他汀按10 mg/kg进行灌胃.于造模后12 h、1 d、2 d、3 d和5 d分别处死各组大鼠留取脑组织.采用免疫组化法和Western blot法检测各组大鼠脑组织TNF-α、IL-1β及NF-κB的蛋白表达情况.结果 与假手术组相比,脑出血组TNF-α、IL-1β和NF-κB的表达增加(P<0.05),均为12 h开始持续增加,3 d时达高峰,持续表达至5 d;与脑出血组相比,治疗组TNF-α、IL-1β和NF-κB的表达减少(P<0.05).结论 脑出血大鼠血肿周围脑组织存在NF-κB、TNF-α和IL-1β的高表达.阿托伐他汀可通过下调NF-κB信号分子的表达及炎性细胞因子的产生,减轻脑出血后脑组织的炎症反应,从而起到脑保护的作用.