NF-kappa B p65和VEGF在原发性肝癌中的表达及意义

[目的]探讨核转录因子(NF-kappa B p65)和血管内皮生成因子(VEGF)在晚期原发性肝癌(HCC)组织和正常肝脏组织中的表达及其意义。[方法]采用免疫组织化学染色SP法检测65例晚期HCC组织切片(其中36例直径≥5cm,16例5cm>直径>2cm,13例直径≤2cm;28例肝内病灶数目大于1个,37例肝内病灶数目为1个)和30例正常肝脏组织(对照组)中NF-kappa B p65和VEGF抗体的表达水平,采用秩和检验方法分析其差异性,并再以直线相关方法分析NF-kappa B p65和VEGF在HCC中表达的相关性。[结果]各肿瘤组随肿瘤直径的增大,NF-kappa B p65和VEGF表达水平均出现递增的趋势。肝内(肿瘤数目>1个)NF-kappa B p65和VEGF表达水平强于肝内无转移组(P<0.05)。NF-kappa B p65和VEGF在HCC中表达呈正相关。[结论]VEGF有促进肝癌生长和侵袭转移作用,而NF-kappaBp65在HCC生长中有促进VEGF表达上调的作用。

非小细胞肺癌中NF-kappa B的表达与凋亡的关系

目的:研究核因子kappa B(nuclear factor kappa B,NF-kappa B)在非小细胞肺癌中的表达,探讨不同NF-kappa B表达的非小细胞的凋亡率,明确NF-kappa B在非小细胞肺癌中的临床意义。方法:应用Western blot检测了45例从2005年10月到2005年12月手术切除的非小细胞肺癌的新鲜标本,并用Tunel法测其凋亡率。结果:在45例患者中,NF-Kappa B的相对量是0.6047±0.3572,而且分化差的非小细胞肺癌中的NF-kappa B的表达比分化好的非小细胞肺癌高(P<0.05);在NF-kappa B高表达中肺癌细胞的凋亡率为56.4%,而在低表达中则为76.7%(P<0.05)。结论:NF-kappa B的表达与非小细胞肺癌的分化相关,在非小细胞肺癌中,NF-kappa B抑制凋亡,并在非小细胞肺癌的形成和发展中起重要作用,或许可以作为基因治疗的靶点为非小细胞肺癌的诊治提供新的思路。

离子硅对成骨细胞NF-kappa B核内转录因子活性的影响

目的研究离子硅对体外培养的成骨细胞NF-kappa B核转录因子活性的影响。方法分别用终浓度为1 mmol/L和2 mmol/L离子硅(SiO32)-处理MC3T3-E1成骨细胞,处理时间分别为6、12、24和48 h,设置对照组(不加处理因素);采用流式细胞术检测细胞周期,计算细胞的增殖指数;Westernblotting方法检测NF-kappa B信号通路的相关蛋白表达量及其变化。结果流式细胞术结果显示,与对照组相比,1 mmol/L浓度的离子硅处理24 h组和48 h组,MC3T3-E1细胞增殖明显;Western blot结果显示,1 mmol/L浓度的离子硅促进成骨细胞增殖与p-NF-kappa B表达上升密切相关。结论骨材料中释放的微量的硅不会引起成骨细胞损伤,相反,微量的硅酸盐可能通过激活NF-kappa B诱导成骨细胞增殖。

NF-kappa B抑制剂PDTC抑制IL-1β诱导软骨细胞生成NO的实验研究

为探讨PDTC对IL 1β诱导的大鼠软骨细胞生成NO的影响 ,寻找抑制NO生成的新途径。分离并培养大鼠软骨细胞 ,加入不同度的PDTC ,以IL 1β刺激软骨细胞 ,RT PCR检测iNOS的mRNA表达 ,NO检测试剂盒检测NO的含量。结果显示PDTC能显著抑制IL 1β诱导的iNOSmRNA的表达及NO的生成且呈剂量依赖性。表明PDTC能抑制IL 1β诱导的NO生成 。

Activation of TLR-4 and liver injury via NF-kappa B in rat with acute cholangitis

BACKGROUND: Toll-like receptors (TLRs) are a family of type 1 transmembrane receptors, which can recognize different pathogen-associated molecular patterns. Among them, TLR-4 is specific to lipopolysaccharide. It transfers the infection signal into the cell and promotes the translocation of nuclear factor kappa B (NF-kappa B) to the nucleus and the subsequent transcriptional activation of genes encoding pro- and anti-inflammatory cytokines and chemokines. Acute cholangitis (AC) is a common biliary tract infection in oriental countries, and often leads to liver injury. The activation of TLR-4 and its significance in liver injury in rats with AC remain unclear. METHODS: Rat models of AC (biliary tract obstruction+E. coli injection, n=36) and control models (biliary tract obstruction+saline, n=18) were made. Liver tissue injury was investigated by pathological examination. The levels of serum TNF-alpha and IL-10 were measured by enzyme-linked immunosorbent assay, and the expressions of TLR-4, NF-kappa B mRNAs and proteins in the liver were detected by RT-PCR, immunohistochemical staining and Western blotting, respectively. RESULTS: Severe liver tissue injury in rats with AC was evident as shown by pathological examination. TLR-4 and NF-kappa B were strongly expressed in the cytoplasm of hepatocytes in the AC group. They were negative or slightly positive in the control group. TLR-4 mRNA and protein in the liver of rats with AC increased 1 hour after biliary tract ligation and E. coli injection, and peaked at 6 hours after surgery. Twenty-four hours later, they began to decrease. The expression of TLR-4 was paralleled by that of NF-kappa B in the liver and TNF-alpha in serum. CONCLUSION: The higher expression of TLR-4 in the liver of rats with AC may be involved in liver injury through the activation of NF-kappa B and release of cytokines such as TNF-alpha.

肺炎衣原体感染和高脂血症对心肌细胞NF-kappa B和AP-1的影响

目的:探讨肺炎衣原体感染和高脂血症对心肌细胞炎症的影响。方法:用间接免疫荧光的方法检测肺炎衣原体感染或给予高脂饮食的C57BL/6J小鼠,观察NF-κB亚单位P50和c-Fos在小鼠心肌细胞中表达程度。结果:肺炎衣原体感染和高脂血症能引起心肌细胞中P50和c-Fos的激活。对照组心肌细胞核中未见P50和c-Fos的表达,而3个实验组心肌细胞核中都有不同程度的P50和c-Fos表达。实验组和对照组比较P<0.01,在3个实验组间无显著差异。结论:在肺炎衣原体感染和高脂血症形成的早期,心肌细胞的炎症通路已被激活。

Effects of Dan-shao-hua-xian on expression of PPAR-gamma and NF-kappa B in rat liver fibrosis

BACKGROUND: Peroxisome proliferator-activated receptor gamma (PPAR-gamma) and nuclear factor kappa B (NF-kappa B) play important roles in liver fibrosis. This study aimed to investigate the effects of Dan-shao-hua-xian, a preparation of traditional Chinese medicine, on the expression of PPAR-gamma and NF-kappa B in the fibrotic livers of rats. METHODS: Seventy Wistar rats were randomly divided into 4 groups: treatment (model, 8 weeks+treatment, 8 weeks; group A), natural recovery (model, 8 weeks+ saline, 8 weeks; group B), model (model only, 8 weeks; group Q, and control (normal, untreated, 16 weeks; group D). Each group consisted of 20 rats (except for group D, which had 10). Fibrotic liver models were induced in rats by subcutaneous injection of CCI4, oral administration of alcohol and a high-lipid/low-protein diet for 8 weeks. After the models were established, the rats in group A were orally given Dan-shao-hua-xian capsules daily for another 8 weeks. Then, the liver indices serum hyaluronic acid (HA), tumor necrosis factor-alpha (TNF-alpha) and alanine aminotransferase (ALT) were measured. The degree of hepatic fibrosis was evaluated by optical microscopy. Hydroxyproline (Hyp) in the liver tissue was determined. The expression of PPAR-gamma was detected by immunohistochemical techniques. The protein levels of PPAR-gamma and NF-kappa B were determined by Western blotting. RESULTS: The concentrations of serum HA, TNF-alpha and Hyp in group C increased compared with group D (P<0.05), and they decreased in group A compared with group C (P<0.05). The expression of PPAR-gamma in group C decreased compared with group D (P<0.05), and it increased in group A compared with groups B and C (P<0.05). Similarly, Western blotting showed that the expression of PPAR-gamma in group C decreased compared with group D, and it increased in group A compared with group C. The expression of NF-kappa B increased in group C compared with group D (P<0.05), and it decreased in group A compared with group C (P<0.05). CONCLUSION: Dan-shao-hua-xian capsules enhance the expression of PPAR-gamma but decrease that of TNF-alpha and NF-kappa B in the liver tissues of CCI4-induced hepatic fibrotic rats. These effects may play a role in its activity in treating hepatic fibrosis.

17β-雌二醇抑制NF-kappa Bp65表达对CCl_4诱导大鼠肝纤维化中肝细胞损伤的意义

目的探索17β-雌二醇(E2)抑制NF-kappa Bp65的表达在四氧化碳(CCl4)诱导Wistar大鼠肝纤维化中肝细胞损伤的意义。方法采用实时荧光定量PCR(SYBR GREEN)技术测量雌二醇治疗的肝纤维化模型大鼠肝脏组织中雌激素受体α(ERα)mRNA和NF-kappa Bp65mRNA的表达;采用免疫组化S-P方法检测肝细胞NF-kappa Bp65和ICAM-1表达并分析与肝细胞损害的关系。结果雌二醇治疗组的肝细胞纤维化模型大鼠ERαmRNA表达高于对照组和他莫昔芬组(P<0.05),而NF-kappa Bp65和ICAM-1表达以及ICAM-1免疫组化表达则低于对照组和他莫昔芬组(P<0.05);雌二醇治疗组的肝细胞损伤的程度也低于对照组和他莫昔芬组。结论E2通过ERα途径抑制NF-kappa Bp65在大鼠肝细胞中的表达,从而减轻了肝细胞的损伤。

Relationship between the Expression of NF-kappa B and Apoptosis in Non-Small Cell Lung Cancer

OBJECTIVE To study the expression of the nuclear factor kappa B (NF-kappa B) in non-small cell lung cancer (NSCLC),to explore the apoptotic ratio in NSCLC related to different NF-kappa Bs,and to understand the clinical significance of NF-kappa B in NSCLC apoptosis. METHODS NF-kappa B expression in 45 new samples of NSCLC,collected during a period from October to December,2005,was assayed using Western blots,and the apoptotic ratio of NSCLC was determined by the Tunel method. RESULTS Of the 45 patients,the average relative expression of NF-kappa B was 0.6047±0.3572.The expression of NF-kappa B was higher in the poorly differentiated lung cancer cells than in the well-differentiated tumors(P<0.05).The apoptotic ratio was 56.4% in the lung cancer cells with higher NF-kappa B expression,and was 76.7% in those with lower NF-kappa B expression(P<0.05). CONCLUSION The expression of NF-kappa B is correlated with the differentiation of NSCLC.NF-kappa B inhibits apoptosis in NSCLC.As a transcription factor,NF-kappa B plays a very important role both in formation and in development of NSCLC. NF-kappa B might serve as a target for NSCLC gene therapy.

NF-kappa B和AP-1在非小细胞肺癌中的表达

背景与目的核因子kappaB(NFkappaB)和激活蛋白1(AP1)在细胞凋亡和增生过程中所起的作用逐渐被人们所认知,在肿瘤的形成过程中也扮演着重要的角色。本研究分析了NFkappaB、AP1在非小细胞肺癌中的表达,以明确二者之间的相互关系,并进一步研究二者对周期蛋白cyclinD1和caspase3在非小细胞肺癌中表达的影响。方法应用Westernblot检测NFkappaB、AP1、cyclinD1和caspase3在非小细胞肺癌中的蛋白表达,应用RTPCR检测不同NFkappaB和AP1表达的肺癌组织中cyclinD1和caspase3的mRNA表达。应用相关分析判断NFkappaB和AP1的相关性。结果在45例非小细胞肺癌患者中,NFkappaB和AP1在肺癌组织中的表达均高于癌旁肺组织中的表达(0.6047比0.2798,P<0.01)。在NFkappaB和AP1较高表达的肺癌组织中,cyclinD1蛋白表达和mRNA表达均增加(P<0.01),而caspase3的蛋白表达和mRNA表达减少(P<0.01)。相关分析显示NFkappaB和AP1有明显的相关性(r=0.800,P<0.01)。结论NFkappaB和AP1作为转录因子可能在非小细胞肺癌的形成和发展中起重要作用。

岩黄连总碱和脱氢阿卟卡维丁对Tca8113中NF-kappa B表达活性的影响

目的:研究中草药岩黄连总碱(YHL-TA)和脱氢阿卟卡维丁(YHL-2)对人舌鳞癌细胞Tca8113细胞中NF-kappaB表达活性的影响,并进一步探讨其抗癌机制。方法:采用PCR法和Western-blotting法检测经YHL-TA和YHL-2作用后的Tca8113细胞中NF-kappa B核酸水平以及蛋白水平的表达活性变化。结果:YHL-TA和YHL-2均能抑制Tca8113细胞增殖,并在核酸水平和蛋白水平抑制NF-kappa B P50、P65亚基的表达活性。其中YHL-2抑制作用最强;并且随作用时间延长和药物浓度的增高,抑制作用增强。结论:YHL-TA和YHL-2可以明显抑制Tca8113细胞的增殖,并显著降低其NF-kappa B的表达,其中YHL-2作用最明显。这可能是岩黄连抗癌作用机制之一。

The Expression of NF-Kappa B Family Protein and Its Relationship with Drug Resistance in Breast Cancer Cell Lines

Objective: To study the expression of NFκB family protein in breast cancer cell lines and the relationship between NFκB family protein and the drug resistance.Methods: Expression of P65, IκB-α in 14 breast cancer cell lines and P50 in 11 breast cancer cell lines was detected by Western blot. The sensitivity of the cells to ADM was determined by MTT.Results: 1κB-α located mainly in the cytoplasm. P65 and P50 were in both of cytoplasm and nucleus. The expression level of P50 was higher than that of P65, especially in nucleus. MTT assay showed that IC50 was three-fold higher in the cell lines which expressed P50 in nucleus than those P50 negative in nucleus, but no difference was found in the expression of P65.Conclusion: The expression of P50 in nucleus may predict the chemotherapy resistance in breast cancer, so it can be used as an indicator to predict the prognosis. The expression of P50 is more often in breast cancer, and it may play a more important role in the chemotherapy resistance than other NFκB family members. Key words breast cancer - NFκB - chemotherapy resistance

PC12细胞转染nNOSα和nNOSβ对NF-kappa B转录活性和细胞凋亡的影响

目的:比较转染nNOSα和nNOSβ的PC12细胞NF-kappaB转录活性和细胞凋亡百分数,探讨nNOSα和nNOSβ在功能上的差异性.方法:将携带nNOSα和nNOSβ基因的pcDNA3.0表达质粒转化感受态细胞,在大肠杆菌中扩增后,提纯质粒酶切鉴定.nNOSα和nNOSβ基因的pcDNA3.0表达质粒通过脂质体转染PC12细胞,G418筛选,使用抗nNOS羧基端的抗体,通过免疫组织化学和WesternBlot方法检测nNOS的表达效率.提取经G418筛选PC12细胞核蛋白,采用凝胶电泳阻滞实验(EMSA)检测细胞核NF-kappa B转录活性.不同PC12细胞经PI染色后流式细胞仪分选,测定不同增殖周期的细胞数.结果:质粒酶切鉴定结果与携带nNOSα和nNOSβ基因的pcDNA3.0质粒结构相符;免疫组织化学和Western Blot方法检测转染nNOS基因后表达的nNOSα和nNOSβ均高于未转染的PC12细胞(P<0.05),PC12细胞转染nNOSα增加NF-kappa B转录活性(P<0.05),而nNOSβ降低NF-kappaB转录活性(P<0.05);转染nNOSα和nNOSβ及未转染的PC12细胞均不改变细胞凋亡百分数(P>0.05).结论:nNOSα增加PC12细胞NF-kappaB转录活性,nNOSβ降低C12细胞NF-kappa B转录活性,但均不改变细胞凋亡百分数,nNOSα和nNOSβ在功能上具有差异性.

Effects of ivabradine on Notch and NF-kappa B signaling pathway in myocardial cells of rats with myocardial infarction

Objective:To investigate the effects of ivabradine on Notch and NF-kappa B signaling pathway in myocardial cells of rats with myocardial infarction.Methods: The model of myocardial infarction was established by ligating the left anterior descending coronary artery. The surviving rats were randomly divided into model group (MI group,n=8) and treatment group (IVA group,n=8). Rats with the same location but without ligation of the left anterior descending coronary artery were used as control group (CON group,n = 8). IVA was administered for 28 d. Hemodynamic and cardiac function indexes of all rats were measured: heart rate (HR), systolic pressure (SBP), diastolic pressure (DBP), mean arterial pressure (MAP), left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP) and maximum rate of increase and decrease of left ventricular internal pressure (+dp/dt);left ventricular mass index, left ventricular cross-sectional diameter and infarct area;The expression of Notch signaling pathway components mRNA (Notch-1, Dll-4, Hes-1) in rat cardiomyocytes was detected by PT-PCR, and the expression of DICD-1 and P65 protein was detected by western-blot. One-way ANOVA was used for comparison between groups, and SNK was used for comparison between groups.Results: SBP, DBP, MAP, LASP, LVEDP and (+dp/dt) in MI group were lower than those in control group (P<0.05), while IVA was higher than those in MI group (P<0.05). Left ventricular mass index and left ventricular sectional diameter in MI group were significantly higher than those in control group (P<0.05), but lower than those in IVA group (P<0.05). The expression of Notch-1 in MI group was significantly higher than that in control group (P<0.05), but lower than that in IVA group (P<0.05). There was no significant difference in the expression of Dll-4 and Hes-1 mRNA between the three groups (P>0.05). The expression levels of NICD-1 and P 65 in MI group were significantly higher than those in CON group (P<0.05), but lower than those in IVA group (P<0.05). Conclusion: IVA may improve cardiac function and inhibit ventricular remodeling in rats with myocardial infarction through Notch and NF-kappa B signaling pathways.

Effects of Oxymatrine on the NF-kappa B expression of HaCaT cells

Objective： To study the effect of oxymatrine on the expression of nuclear factor-kappa B（NF-K B） of Human benign epidermal keratinocytes line（HaCaT cells）. Methods：HaCaT cells were cultured with different concentration of Oxymatrine（10 μ g/ml, 50 μ g/ ml and 100 μg/ml） for 24 h, then 10.5 mol/L Substance P was added to the cells. After 30 min, NF-K B expression in the cells was observed by immunocytochemistry, NF-K B P65 protein expression was evaluated by Western blot, and the mRNA expression of NF-K B P65 was evaluated by reverse transcription polymerase chain reaction（RT-PCR）. The 10.5 mol/L Substance P and culture medium were added to the Substance P group and control group, respectively. Results：In control group, expression rate of positive cells, the expressions of protein and mRNA of NF-K B were all low. In Substance P group, when 10μ mol/L Substance P was added, the expressions were all increased（P 〈 0.05）. But in Oxymatrine groups, the expression rate of positive cells, the expressions of protein and mRNA were all descended in a concentration-dependent manner（P 〈 0.05 or P 〈 0.01）. Conclusion：Oxymatrine can down-regulate the expression of NF-K B of the HaCaT cells and may play an important role in regulating anti-inflammation and immunity.

Ketamine suppresses intestinal NF-kappa B activation and proinflammatory cytokine in endotoxic rats

AIM: To investigate the protective effect of ketamine on the endotoxin-induced proinflammatory cytokines and NFkappa B activation in the intestine. ETHODS: Adult male Wistar rats were randomly divided into 6 groups: (a) normal saline control, (b) challenged with endotoxin (5 mg/kg) and treated by saline, (c) challenged with endotoxin (5 mg/kg) and treated by ketamine (0.5 mg/kg), (d) challenged with endotoxin (5 mg/kg) and treated by ketamine (5 mg/kg ), (e) challenged with endotoxin (5 mg/kg) and treated by ketamine (50 mg/kg), and (f) saline injected and treated by ketamine (50 mg/kg). After 1, 4 or 6 h, TNF-α and IL-6 mRNA were investigated in the tissues of the intestine (jejunum) by RT-PCR. TNF-α and IL-6 were measured by ELISA. We used electrophoretic mobility shift assay (EMSA) to investigate NF-kappa B activity in the intestine. RESULTS: NF-kappa B activity, the expression of TNF-α and IL-6 were enhanced in the intestine by endotoxin. Ketamine at a dose of 0.5 mg/kg could suppress endotoxininduced TNF-α mRNA and protein elevation and inhibit NFkappa B activation in the intestine. However the least dosage of ketamine to inhibit IL-6 was 5 mg/kg in our experiment. CONCLUSION: Ketamine can suppress endotoxin-induced production of proinflammatory cytokines such as TNF-α and IL-6 production in the intestine. This suppressive effect may act through inhibiting NF-kappa B.

Different effects of antisense IRAK-2 oligonucleotide on IL-1 and TNF-stimulated NF-kappa B activation

Objective: To investigate the role of interleukin-1 (IL-1) receptor associated kinase-2 (IRAK-2) in IL1 and TNF-stimulated nuclear factor-kappa B (NF-kappa B ) activation. Metbods: The 293 cell was trans fectedwith antisense IRAK-2 oligonucleotide (IRAK-2 ODN) followed by stimulating the cell with IL-1 or tumor necrosis factor (TNF), and then the levels of NF-kappa B activation was analyzed by sandwich enzyme-linked immunosorbent assay (ELISA ). Result: Pre-transfecting with antisense IRAK-2 ODN could remarkably decreasethe levels of NF-kappa B activation stimulated by IL-1 in time- and concentration-dependent manner, it can not attenuate the one stimulated by TNF. Conclusion: The responses of IL-1 and TNF-stimulated NF-kappa B activation to antisense IRAK-2 oligonucleotids were different. IRAK-2 plays a key role in the IL-1 signaling events leading to NF kappa B activation.

Effect of dexmedetomidine on the prevention of PSH in patients with severe craniocerebral injury by regulating TLR4/My D88/NF-kappa B signaling pathway

Objective:To investigate the clinical efficacy of dexmedetomidine in the regulation of TLR4/My D88/NF-κB in the prevention of paroxysmal sympathetic over-excitation (PSH) in patients with severe head injury. Methods:One hundred patients with severe head injury who were admitted to our hospital from September 2016 to May 2019 were enrolled. The randomized digital table method was divided into 50 cases in the study group and the control group. Patients in the study group were given dexmedetomidine at a dose of 1.0 μg/kg before anesthesia induction, followed by infusion at 0.4 μg / (kg·h), and the control group was injected with the same amount of normal saline. The incidence of PSH, clinical symptoms, imaging findings, mechanical ventilation time, tracheal intubation/incision duration, ICU hospitalization time, total length of hospital stay, and GCS scores three months after discharge were compared between the two groups. At the same time, the fluorescence intensity, TLR4, NF-κB expression level and tumor necrosis factor-α (TNF-α) expression levels in peripheral blood CD14+ monocytes of the two groups were detected. Results:The incidence of PSH was significantly lower in the study group than in the control group at 7 and 3 months (P<0.05). The total length of hospital stay, duration of ICU hospitalization, intraoperative tracheotomy, and mechanical ventilation time were significantly lower in the study group than in the control group. And the GCS score was higher than the control group, and the difference was statistically significant (P<0.05). In addition, the imaging results showed that there were some differences in the location of imaging lesions between the two groups. The proportion of lesions in the ventricular system and surrounding areas was higher in the control group than in the study group (P<0.05). And the T14-T3 CD14+ PBMC MyD88 fluorescence intensity, TLR4 and NK-κB positive expression rate were significantly higher than those of T0 (P<0.05), but the MyD88 fluorescence intensity, TLR4 and NK-κB positive expression rate in the study group were significantly lower than those in the control group at T1~T3 (P<0.05). The levels of serum TNF-α in T1~T3 groups were significantly higher than those in T0 (P<0.05), but the levels of serum TNF-α in T1~T3 in the study group were significantly lower than those in the control group (P< 0.05). Conclusions:Dexmedetomidine can reduce the oxidative stress response in patients with severe head injury by inhibiting TLR4/My D88/NF-κB signaling pathway, thus effectively reducing the risk of PSH and improving the prognosis of patients.