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。

Suppressing high mobility group box-1 release alleviates morphine tolerance via the adenosine5'-monophosphate-activated protein kinase/heme oxygenase-1 pathway

Opioids,such as morphine,are the most potent drugs used to treat pain.Long-term use results in high tolerance to morphine.High mobility group box-1(HMGB1) has been shown to participate in neuropathic or inflammatory pain,but its role in morphine tolerance is unclear.In this study,we established rat and mouse models of morphine tolerance by intrathecal injection of morphine for 7 consecutive days.We found that morphine induced rat spinal cord neurons to release a large amount of HMGB1.HMGB1 regulated nuclear factor κB p65 phosphorylation and interleukin-1β production by increasing Toll-like receptor 4receptor expression in microglia,thereby inducing morphine tolerance.Glycyrrhizin,an HMGB1 inhibito r,markedly attenuated chronic morphine tole rance in the mouse model.Finally,compound C(adenosine 5’-monophosphate-activated protein kinase inhibitor) and zinc protoporphyrin(heme oxygenase-1 inhibitor)alleviated the morphine-induced release of HMGB1 and reduced nuclear factor κB p65 phosphorylation and interleukin-1β production in a mouse model of morphine tolerance and an SH-SY5Y cell model of morphine tole rance,and alleviated morphine tolerance in the mouse model.These findings suggest that morphine induces HMGB1 release via the adenosine 5’-monophosphate-activated protein kinase/heme oxygenase-1 signaling pathway,and that inhibiting this signaling pathway can effectively reduce morphine tole rance.

硫化氢抑制p38MAPK和NF-κBp65信号通路活性改善糖尿病心肌纤维化的作用和机制

目的研究外源性硫化氢(H2S)对糖尿病心肌病小鼠心肌纤维化的改善作用及其机制。方法 24只雄性C57BL/6J小鼠随机分为正常对照组(NC组)、糖尿病对照组(ALX组)、低剂量Na HS治疗组(ALX+LDNa HS组)、高剂量Na HS治疗组(ALX+HDNa HS组),每组6只。采用四氧嘧啶(ALX)200mg/kg腹腔注射建立糖尿病小鼠模型,造模成功后,NC组和ALX组每天自由饮用自来水,ALX+LDNa HS组和ALX+HDNa HS组自由饮用浓度分别为30μmol/L和90μmol/L的Na HS(H2S供体)溶液。喂养8周后取材,采用HE染色观察心肌组织形态学变化,Real-time PCR检测p38丝裂原活化蛋白激酶(p38MAPK)、核转录因子-κB p65(NF-κB p65)m RNA的表达,Western blotting检测p-p38MAPK、p-NF-κB p65和Ⅰ型胶原蛋白的表达。结果与NC组比较,ALX组心肌组织胶原表达量增加,心肌间质出现纤维化,p38MAPK、NF-κB p65 m RNA表达明显增加(P<0.01),同时p-p38MAPK、p-NF-κB p65和Ⅰ型胶原蛋白表达也明显增高(P<0.01)。经硫化氢干预后糖尿病小鼠心肌纤维平行排列,心肌组织胶原表达量明显减少,心肌间质有少量结缔组织;p38MAPK、NF-κB p65 m RNA表达明显减少(P<0.01),p-p38MAPK、p-NF-κB p65和Ⅰ型胶原蛋白表达明显下降(P<0.01),且ALX+HDNa HS组较ALX+LDNa HS组明显改善(P<0.05)。结论 H2S可改善糖尿病小鼠心肌纤维化,可能与调节p38MAPK和NF-κB p65介导的炎症反应有关。

Protective effect of Jiaweibugan decoction against diabetic peripheral neuropathy

Oxygen free radical damage is regarded as a direct or indirect common pathway associated with diabetic neuropathy and is the main cause of complications in peripheral neuropathies. We speculate that Jiaweibugan decoction has a significant effect in treating diabetic peripheral neuropathy through an anti-oxidative stress pathway. In this study, a diabetic rat model was established by intraperitoneal injection of streptozotocin. Rats were treated with Jiaweibugan decoction via intragastric administration. The levels of malondialdehyde and glutathione, which are indirect indexes of oxidative stress, in serum were determined using a colorimetric method. The expression levels of nuclear factor kappa B p65 mRNA and p38 mitogen-activated protein kinase, which are oxidative stress associated factors, in the dorsal root ganglion of spinal $4-6 segments were evaluated by reverse-transcriptase polymerase chain reaction and immunohistochemistry. Results showed that, Jiaweibugan decoction significantly ameliorated motor nerve conduction velocity in diabetic rats, effectively decreased malondialdehyde levels in serum and the expression of nuclear factor kappa B p65 mRNA and p38 mitogen-activated protein kinase mRNA in the dorsa root ganglion, and increased glutathione levels in serum. Therefore, our experimental findings indicate that Jiaweibugan decoction plays an anti-oxidative stress role in the diabetic peripheral neuropathy process, which has a protective effect on peripheral nerve injury.

NF-κB和ICAM-1在喉癌中的表达及其意义

目的 :研究体内核转录因子NF -κB和细胞间粘附分子ICAM - 1在喉癌组织中表达及意义。方法 :采用免疫组织化学染色S -P法检测 4 0例喉癌组织和 2 0例正常成年人喉粘膜组织的NF -kappaBP6 5蛋白及细胞间粘附分子 -I(ICAM -1)的表达水平。结果 :(1)与正常喉粘膜组织相比较 ,喉癌组织中NF -kappaBP6 5蛋白和ICAM - 1表达水平均呈显著性增强 (P <0 .0 0 1)。 (2 ) 4 0例喉癌组织检测数据提示NF -kappaB蛋白表达和ICAM - 1表达呈直线回归关系 (b =0 .975 ,P <0 .0 0 1)。结论 :(1)NF -kappaB可能参与喉癌发生发展机制 ;(2 )NF -kappaB的激活导致喉癌患者体内ICAM - 1表达增强 ,促进了机体自身细胞免疫抗肿瘤的作用。

核因子-κB/p65表达下调对舌鳞状细胞癌裸鼠移植瘤中bcl-2和bax表达的影响

目的:本实验旨在探讨下调核因子-κB(nuclear factor-kappa B,NF-κB)/p65亚单位的表达对舌鳞状细胞癌裸鼠移植瘤生长及细胞凋亡相关基因bcl-2和bax表达的影响。方法:应用舌鳞状细胞癌Tca8113细胞建立舌鳞状细胞癌裸鼠移植瘤模型。将针对p65的小干扰RNA(small interfering RNA,siRNA)接种荷瘤裸鼠,观察下调NF-κB/p65表达后荷瘤裸鼠的生长情况;采用TUNEL法检测肿瘤组织中的细胞凋亡情况;进一步应用RT-PCR和Western印迹法检测肿瘤组织中p65、bcl-2和bax的mRNA及蛋白表达变化。结果:经p65siRNA治疗后,肿瘤的生长明显受到抑制(P<0.05);TUNEL结果表明,p65siRNA组肿瘤细胞明显发生凋亡;此外,p65siRNA组裸鼠肿瘤组织p65和bcl-2的mRNA及蛋白表达明显下调,而bax的mRNA及蛋白表达显著上调,差异均有统计学意义(均P<0.05)。结论:NF-κB/p65亚基可能在舌鳞状细胞癌的细胞凋亡中发挥重要作用。

NF-κB/p65在鼻NK／T淋巴瘤中的表达及意义

目的探讨NF-κB／p6S在鼻NK／T淋巴瘤表达及其与细胞凋亡和预后的关系。方法应用免疫组织化学SP法和TUNEL法，检测23例鼻NK／T淋巴瘤和14例良性淋巴结病变中NF-κB／p6S的表达水平和细胞凋亡。结果NF-κB／p65在鼻NK／T淋巴瘤表达的阳性率为43．5％（10／23）；凋亡指数与NF-κB／p65呈负相关（r=-0．631，P〈0．05）。NF-κB/p65阳性表达者的平均生存时间明显短于阴性者（P〈0．05）。结论NF-κB／p65表达而引起的凋亡抑制可能在鼻NK／T淋巴瘤的发生发展中有重要作用并可作为预后不良的参考指标。

磷酸酶基因、核因子-κBp65、Survivin蛋白在子宫内膜样腺癌中的表达及其临床意义

目的探讨磷酸酶基因（PTEN）、核因子（NF-κB p65）及凋亡抑制蛋白Survivin基因在子宫内膜样腺癌中的表达及其在子宫内膜样腺癌的发生、发展中的作用。方法采用SP免疫组织化学法检测63例子宫内膜样腺癌、20例正常增生期子宫内膜组织中PTEN、NF-κB p65及Survivin蛋白的表达。结果子宫内膜样腺癌与正常增生期子宫内膜组织中PTEN、NF-κB p65及Survivin蛋白表达的阳性率差异有统计学意义（P〈0．01）。PTEN表达与子宫内膜样腺癌分化程度呈正相关（P〈0．001），与淋巴结转移及TNM分期呈负相关（P〈0．005），NF-κB p65的表达与子宫内膜样腺癌分化程度呈负相关（P〈0．05），与浸润深度、淋巴结转移及TNM分期呈正相关（P〈0．05）。Survivin与子宫内膜样腺癌与浸润深度、淋巴结转移及TNM分期呈正相关（P〈0．05）。结论PTEN、NF-κB p65及Survivin在子宫内膜腺癌的发生、发展中起着不同程度的作用，联合检测PTEN、NF-κB p65及Survivin对研究子宫内膜腺癌的发生、发展、早期诊断及预后评估有一定的参考价值。

Pingchuan formula(平喘方) improves allergic asthma in mice through inhibiting nuclear factor-kappa B/mitogen-activated protein kinase signaling pathway

OBJECTIVE:To examine the role and decipher the mechanism of Pingchuan formula(平喘方,PCF)in treating allergic asthma.METHODS:The mice were treated with saline,dexamethasone(DXM)and PCF for 1 week after the asthma model was established and their respiratory function including respiratory resistance(RI),pulmonary dynamic compliance(Cdyn)and maximum voluntary ventilation(MVV)were measured.In addition,cellular changes in bronchoalveolar lavage fluid(BALF)and pathological changes in lung biopsy as well as the expression level ofα-smooth muscle actin(α-SMA),transforming growth factor-beta1(TGF-β1)in BALF and interleukin-5(IL-5),interleukin-13(IL-13),tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ),nuclear factor-kappa B-p65(NF-κBp65),inhibitor-αof nuclear transcription factorκB(IκBα),p38 mitogen-activated protein kinase(p38 MAPK),c-jun n-terminal kinase(JNK)and its phosphorylated proteins in lung tissue were also examined and compared among different groups.RESULTS:Our data suggested that the respiratory functions were significantly improved and the pathological changes ameliorated in the DXM group and the PCF group compared to the model group.Both DXM and PCF effectively decreased the number of eosinophils,lymphocytes,and neutrophils in BAL as well as the secretion ofα-SMA and TGF-β1,IL-5,IL-13,while increased the expression of TNF-αand IFN-γ.Furthermore,our study indicated that the NF-κBp65,IκBα,p38 MAPK and JNK pathways were inhibited under the treatment of PCF.CONCLUSION:Our data indicated that PCF can attenuate the inflammatory response in asthma through inhibiting the NF-κB/MAPK signaling pathway.This study not only supported the use of PCF in allergic asthma in clinic but also shed light upon afurther understanding of the disease pathogenesis.

川芎嗪对土三七诱导的小鼠肝小静脉闭塞病治疗机制的研究

背景:肝小静脉闭塞病(HVOD)临床上以肝肿大、黄疸、腹水和体重增加为特征,目前尚缺乏有效的治疗手段。本课题前期研究发现川芎嗪对土三七诱导的小鼠HVOD具有治疗作用。目的:探讨川芎嗪对土三七诱导的小鼠HVOD的治疗机制。方法:将115只小鼠随机分为土三七组(土三七浓缩煎液30 g·kg^(^(-1))·d^(-1)灌胃)、低剂量川芎嗪干预组(土三七浓缩煎液30 g·kg^(-1)·d^(-1)+川芎嗪100 mg·kg^(-1)·d^(-1)灌胃)、高剂量川芎嗪干预组(土三七浓缩煎液30 g·kg^(-1)·d^(-1)+川芎嗪200 mg·kg^(-1)·d^(-1)灌胃)和正常对照组(PBS 30 g·kg^(-1)·d^(-1)灌胃),30 d后处死所有小鼠。行HE染色和Masson染色,以RT-PCR法和蛋白质印迹法分别检测肝组织中组织因子(TF)、核因子(NF)-κBp65、早期生长反应因子^(-1)(Egr^(-1))mRNA和蛋白表达。结果:HE染色和Masson染色结果显示川芎嗪可明显改善HVOD小鼠肝组织病理损伤。土三七组TF、NF-κBp65和Egr^(-1) mRNA和蛋白表达显著高于正常对照组,组间差异有统计学意义(P<0.05);川芎嗪干预后,TF、NF-κBp65和Egr^(-1) mRNA和蛋白表达均不同程度下降(P<0.05),以高剂量组下降更为明显,而高剂量川芎嗪干预组与正常对照组相比差异无统计学意义(P>0.05)。结论:川芎嗪可能通过下调NF-κBp65和Egr^(-1)表达降低TF水平,阻止凝血系统活化,从而有效治疗HVOD,且高剂量川芎嗪的疗效更为确切。

Human papillomavirus, tobacco, and poor oral hygiene can act synergetically, modulate the expression of the nuclear factor kappa B signaling pathway for the development and progression of head and neck cancer in the Pakistani population

Background: Head and neck cancers (HNCs) are a heterogeneous group of tumors that progress owing to varied enviromental and genetic risk factors. Viral infections are threatening and adept at altering the expression of cellular transcription factors such as nuclear factor kappa B (NF-κB) and deregulation of other cellular proteins like NF kappa B inhibitor alpha (IκBα). The present study was conducted to detect high-risk genotypes of human papillomavirus (HPV) and protein expression of NF-κB signaling pathway in HNC patients with HPV infection.Methods: For HPV detection, genomic DNA from 152 HNC tumors was extracted formalin-fixed paraffin-embedded tissue DNA kit. For genotyping, polymerase chain reaction (PCR) using a general primer, HPV type-specific primers and agarose gel electrophoresis were performed. Immunohistochemistry (IHC) was also performed on 4-μm thick tissue sections using HPV E6 monoclonal antibody. Protein expression analysis of NF-κB signaling pathway including p50, p65, and IκBα was performed using IHC.Results: PCR analysis showed that 24.3% (37/152) of HNC cases were HPV positive. Among HPV positive, 86.5% (32/37) were tobacco users, while among HPV negative, 66.9% (77/115) were tobacco users. A significant association of HPV positivity and tobacco user was observed by univariate analysis [P < 0.01;odds ratio (OR): 0.310, 95% confidence interval (CI): 0.110 to 0.870]. More HPV positive patients were with poor oral hygiene (78.3%) when compared with patients with good oral hygiene (21.6%) [P < 0.03, OR: 2.440, 95% CI: 1.650 to 3.600]. The results of the logistic regression analysis showed that age, tobacco use and oral hygiene are significant predictors (P < 0.02). PCR and IHC staining results confirmed that HPV16 was predominant among HNC cases (64.8%) when compared with HPV18 (35.2%). Expression of NF-κB proteins (p50, p65, and IκBα inhibitor) were also observed in HPV and non-HPV infected HNC tissues. IHC expression of p50, and p65 showed nuclear staining, while IκBα inhibitor showed cytoplasmic staining. Protein expression in HPV cases was higher as compared to HPV naive cases (P < 0.05).Conclusions: From the study, it can be established that the use of tobacco, oral hygiene, and HPV infection may be synergistically involved in modulating the expression of NF-κB signaling pathway for the development and progression of HNC in the Pakistani population.

肺炎克雷伯菌肺炎对小鼠NLRP3炎症通路的影响

目的建立肺炎克雷伯菌肺炎小鼠模型,并探究肺炎克雷伯菌感染对核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症通路的影响。方法给小鼠气管内接种肺炎克雷伯菌,观察小鼠死亡率;接种肺炎克雷伯菌48 h后,取小鼠肺部匀浆涂板检查菌落数;取小鼠支气管肺泡灌洗液(BALF)离心,酶联免疫吸附实验(ELISA)检测无细胞上清中白细胞介素(IL)-1β、肿瘤坏死因子(TNF-α)、IL-6、趋化因子配体1(KC)的水平,迈格林华-姬姆萨染色计数细胞沉淀中的中性粒细胞数;取肺部病理切片并进行苏木精-伊红(HE)染色观察肺部病变;提取肺部总蛋白,蛋白质印迹法检测NLRP3、核因子(NF)-κB p65、凋亡相关微粒蛋白(ASC)、天冬氨酸特异性半胱氨酸蛋白水解酶(caspase-1)、人白介素-1β前体(pro-IL-1β)表达水平。结果成功建立肺炎克雷伯菌肺炎小鼠模型,小鼠在144 h内死亡率达到百分之百,肺部有大量细菌负载,肺部中性粒细胞浸润增加,支气管肺泡灌洗液中IL-1β、TNF-α、IL-6、KC分泌升高,肺组织IL-1β、p-p65表达升高。结论通过将肺炎克雷伯菌液接种到小鼠肺部的方法所建立的肺炎小鼠模型稳定性高、可重复性好,肺部NLRP3炎症通路反应强烈。