反义白介素—1受体相关激酶—1;寡核苷酸抑制白介素—1诱导的NF—kB活化

目的：研究反义白素－１受体相关激酶－１（ＩＲＡＫ－１）寡核苷酸对核因子ｋＢ－（ＮＦ－ｋＢ）活化的影响。方法：Ｌｉｐｏｆｅｃｔｉ介导ＩＲＡＫ－１寡核苷酸转染;ｅｐＧ２细胞,以逆转录ＰＣＲ法检测ＩＲＡＫ－１ｍＲＡＮ表达水平,用ＳａｎｄｗｉｃｈＥＬＩＳＡ法检测ＮＦ－ｋＢ的活化。结果;反义ＩＲＡＫ－１寡核苷酸抑制ＩＲＡＫ－１ｍＲＮＡ表达,反义ＩＲＡＫ－１ｍＲＡＮ表达,反义ＩＲＡＫ－１革酸呈剂量（１－８μｇ     

白介素-1受体相关激酶-2反义寡核苷酸对白介素-1和肿瘤坏死因子激活核因子-κB的不同影响

目的　研究白介素 1受体相关激酶 2 (IRAK 2 )在白介素 1(IL 1)与肿瘤坏死因子 (TNF)激活核因子 κB(NF κB)过程中的作用。方法　IRAK 2反义寡核苷酸转染人胚肾细胞 (2 93细胞 ) ,阻断IRAK 2的表达 ,用IL 1及肿瘤坏死因子刺激细胞后 ,用夹心ELISA方法检测NF κB活性。结果　以IRAK 2反义寡核苷酸预处理可明显减弱IL 1诱导的NF κB活性 ,此效应强度存在时间和浓度依赖性 ,但I RAK 2反义寡核苷酸对肿瘤坏死因子诱导的NF κB活性无影响。结论　IRAK 2反义寡核苷酸对IL 1和TNF激活NF κB有不同影响。IRAK 2在IL 1诱导的NF κB信号转导通路中起关键作用     

白介素—1受体相关激酶—2的反义寡脱氧核苷酸体外抑制白介素—1诱导的核因子—κB活化

AIM: To study the inhibitory effects of antisense interleukin-1 receptor associated kinase-2 (IRAK-2) oligodeoxynucleotide (ODN) on interleukin-1 (IL-1)-stimulated nuclear factor-kappa B (NF-kappa B) activation. METHODS: Antisense IRAK-2 ODN was delivered by lipofectin encapsulation into human embryonic kidney 293 cells (HEK 293 cells). The levels of NF-kappa B were assayed by sandwich ELISA. RESULTS: (1) Treatment of HEK 293 cells with IL-1 enhanced NF-kappa B level in nuclei by 518.5% +/- 2.1%. (2) Antisense IRAK-2 ODN inhibited IL-1-induced NF-kappa B activation in a concentration (1-8 micrograms)- and time (5-24 h)-dependent manner. A maximum inhibition was 70.7% +/- 1.0% from Acontrol 0.834 +/- 0.014 to 0.244 +/- 0.008 after treatment with antisense IRAK-2 ODN 4 micrograms for 8 h. CONCLUSION: Antisense IRAK-2 ODN inhibited IL-1-induced NF-kappa B activation.     

反义白介素-1受体相关激酶-1寡核苷酸抑制白介素-1诱导的NF-κB活化

目的 :研究反义白介素 1受体相关激酶 1(IRAK 1)寡核苷酸对核因子 κB(NF κB)活化的影响。方法 :Lipofectin介导反义IRAK 1寡核苷酸转染HepG2细胞 ,以逆转录PCR法检测IRAK 1mRNA表达水平 ,用SandwichELISA法检测NF κB的活化。结果 :反义IRAK 1寡核苷酸抑制IRAK 1mRNA表达 ;反义IRAK 1寡核苷酸呈剂量 (1～ 8μg)和时间 (5～ 2 4h)依赖性地抑制NF κB活化 ,反义IRAK 1寡核苷酸 4μg与细胞共孵育 8h时抑制效果最好。结论 :反义IRAK 1寡核苷酸通过阻断IRAK 1表达抑制NF κB活化 ,预示反义IRAK 1寡核苷酸可潜在抑制IL 1的炎症活性。     

反义磷脂酰肌醇3—激酶寡脱氧核苷酸抑制白介素—1诱导的NF …

目的 研究反义磷脂酰肌醇３－激酶（ＰＩ ３－激酶）寡脱氧核苷酸（ｏｌｉｇｏｄｅｏｘｙｎｕｃｌｅｏｔｉｄｅｓ,ＯＤＮ）对核因子－ｋＢ（ＮＦ－ｋＢ）活化的影响。方法 Ｌｉｐｏｆｅｃｔｉｎ介导反义ＰＩ ３－激酶ＯＤＮ转染ＨｅｐＧ２细胞,以逆转录ＰＣＲ法检测ＰＩ ３－激酶ｍＲＮＡ表达水平,用Ｓａｎｄｗｉｃｈ ＥＬＩＳＡ法检测ＮＦ－ｋＢ的活化。结果 （１）反义ＰＩ－３激酶ＯＤＮ抑制ＰＩ－３激酶ｍＲＮＡ表达。     

白介素-1受体相关激酶-2的反义寡脱氧核苷酸体外抑制白介素-1诱导的核因子-кB活化(英文)

目的:研究白介素-1受体相关激酶-2的反义寡脱氧核苷酸对核因子-кB的抑制作用。方法:脂质体介导反义白介素-1受体相关激酶-2寡脱氧核苷酸转染人胚肾293细胞。夹心酶联免疫吸附测定法检测核因子-кB的含量。结果:(1)白介素-1刺激人胚肾293细胞时,核因子-кB含量明显上升(上升幅度518.5％±2.1％)。(2)白介素-1受体相关激酶-2的反义寡脱氧核苷酸呈浓度(1-8μg)和时间(5-24h)依赖性地抑制白介素-1的作用。反义寡脱氧核苷酸4μg与细胞孵育8h,约70.7％±1.0％的NF-кB活性被抑制。吸光度由对照的0.834±0.01下降到0.244±0.008。结论:白介素-1受体相关激酶-2的反义寡脱氧核苷酸抑制白介素-1诱导的核因子-кB活化。     

白细胞介素-1受体相关激酶-2反义寡核苷酸对IL-1和TNF诱导PGI_2合成的影响(英文)

目的:研究白细胞介素-1受体相关激酶-2(IRAK-2)反义寡核苷酸对白细胞介素-1(IL-1)和肿瘤坏死因子(TNF)诱导前列环素(PGI_2)合成的不同影响。方法:IRAK-2反义寡核苷酸(IRAK-2 ODN)转染脐静脉内皮细胞以阻断IRAK-2的表达,以IL-1及TNF刺激细胞后,用竞争ELISA方法检测PGI_2的合成。结果:IRAK-2 ODN可显著降低IL-1诱导的PGI_2合成,此效应强度存在时间和浓度依赖性,但IRAK-2ODN对TNF诱导的PGI_2合成无影响。结论:IRAK-2 ODN对IL-1和TNF诱导PGI_2合成有不同影响。IRAK-2在IL-1诱导的PGI_2合成中起重要作用。     

脂质体转染bcl—2反义寡核苷酸G3139在HL—60细胞中的动力？…

目的 应用阳离子脂质体介导或直接转染时,建立ＨＬ－６０细胞摄入由５′－ＦＩＴＣ标记的ｂｃｌ－２反义寡核苷酸Ｇ３１３９的动力学模式及观察Ｇ３１３９在细胞内的分布。方法 流式细胞仪测定细胞内的相对平均荧光强度,荧光显微镜观察细胞内的荧光分布。结果 ①脂质体介导转染法显著提高ＨＬ－６０细胞对Ｇ３１３９的摄入,当ＤＯＳＰＥＲ／Ｇ３１３９（ｕｇ／ｕｇ）为２．２／１时,细胞内相对平均荧光强度可达Ｇ３１３９直接     

磷脂酰肌醇3—激酶调控白介素—18诱导核因子—κB活化

目的：研究磷脂酰肌醇（PI）3－激酶在白介素18（IL－18）诱导核因子－κB（NF－κB）活化中的作用。方法：Lipofectin介导反义PI 3－激酶寡核苷酸转染HepG2细胞。用逆转录PCR法检测PI 3－激酶mRNA表达水平,以Sandwich ELISA法检测NF－κB的活化。结果：1）反义PI 3－激酶寡核苷酸抑制PI 3－激酶mRNA表达。（2）IL－18诱导NF－κB活化。（3）反义PI 3－激酶寡核苷酸呈时间（5－24h)和浓度（1－8mg/L)依赖性地抑制IL－18诱导的NF－κB活化。结论：PI 3－激酶调控白介素－18诱导的NF－κB活化。     

Retinoic acid inhibits IL-1-induced IL-6 synthesis in epithelial cells

The regulation of interleukin-6 (IL-6) expression by IL-1 and retinoic acid (RA) was studied in the human epidermoid carcinoma cell line, HEp-2. IL-6 synthesis in HEp-2 cells was induced by IL-1, phorbol myristate acetate (PMA) and calcium ionophore, A23187, but not by dibutyryl-cAMP. RA at 10 mol/L almost completely inhibited IL-1-induced IL-6 production in HEp-2 cells and was less effective in inhibiting the effects of PMA and A23187. Maximal inhibitory effects of RA were observed after 3 days preincubation. To characterize mechanisms involved with the RA effects, we analysed the DNA binding proteins, AP-1, a known target for RA, as well as nuclear factor (NF) IL-6, and NF kB which are involved with the induction of IL-6. Nuclear extracts of HEp-2 cells showed that IL-1 increased only NF kB binding activity and did not change the levels of constitutively expressed AP-1 and NF IL-6. RA did not affect the IL-1-mediated increase in NF kB nor the constitutive levels of AP-1 and NF IL-6 under conditions where it effectively inhibited IL-1-induced IL-6 expression. This demonstrates that RA does not inhibit IL-6 induction via downregulation of IL-1 receptors or inhibition of IL-1 signal transduction pathways. RA might act by inducing other inhibitory factors or by blocking other not yet identified factors essential for IL-1-induced IL-6 synthesis.     

阿托伐他汀对兔动脉粥样硬化中NF-κB/IκB和炎症因子表达的影响

目的探讨高脂饮食诱发兔动脉粥样硬化中核因子-κB(NF-κB)的活化与其抑制因子IκB的表达,以及阿托伐他汀对NF-κB/IκB信号途径、ICAM-1和P选择素的影响。方法24只新西兰♂家兔随机分为正常对照组、高胆固醇组和阿托伐他汀组,应用W estern b lot方法检测动脉中胞核NF-κB p65亚基和胞浆IκBα表达变化;免疫放射法检测血清P选择素水平,免疫组织化学检测血管壁ICAM-1的表达。结果高胆固醇组与对照组比较胞核NF-κB p65和血管壁ICAM-1表达明显增强、血P选择素明显升高(P<0.05),胞浆中IκBα表达明显减弱(P<0.05);阿托伐他汀组与高胆固醇组比较NF-κB p65和血管壁ICAM-1明显表达较弱、血P选择素明显减少(P<0.05),胞浆IκBα的表达增强(P<0.05)。结论NF-κB/IκB信号途径在动脉粥样硬化中起重要作用,阿托伐他汀可以减少P选择素、ICAM-1的表达。     

Salvia miltiorrhiza compounds protect the liver from acute injury by regulation of p38 and NFκB signaling in Kupffer cells

Context: Salvia miltiorrhiza Bunge is a traditional Asian medicine used to treat cerebral and cardiac ischemia. However, the effects of the active compounds of S. miltiorrhiza on liver damage are unclear.

Objective: In this study, we tested the effects on acute liver injury of crude S. miltiorrhiza extracts from roots as well as neotanshinone B, dehydromiltirone, tanshinol A, tanshinone I, dihydrotanshinono I, neotanshinone A, cryptanshinono, tanshinone II A, and salvianolie acid B from purified S. miltiorrhiza extracts.

Materials and methods: Various compounds or ethanol extract of S. miltiorrhiza (50, 100, and 200?mg/kg, p.o.) were administered to rats for five consecutive days. After acute carbon tetrachloride (CCl₄)-induced liver injury by treatment of rats with a single dose of CCl₄ (0.75?mL/kg, p.o), rat liver function was tested by measuring serum biochemical parameters. Serum cytokine concentrations were assessed by enzyme-linked immunosorbent assay (ELISA). Expression of p38 and NFκB was evaluated by western blot.

Results: All S. miltiorrhiza components showed their effects on liver function from the dose from 50 to 200?mg/kg. At the dose of 200?mg/kg, they reduced serum levels of alkaline phosphatase (ALP) by 34–77%, alanine aminotransferase (ALT) by 30–57%, aspartate aminotransferase (AST) by 43–72%, creatine total bilirubin (BIL-T) by 33–81%, albumin (ALB) by 37–67%, indicating that S. miltiorrhiza extracts protected liver from CCl₄-induced damage. Moreover, S. miltiorrhiza extracts at 200?mg/kg reduced the increase in the proinflammatory cytokines tumor necrosis factor-α (TNF-α) by 25–82%, interleukin-1 (IL-1) by 42–74% and interleukin-6 (IL-6) by 67–83%, indicating an effect on alleviating liver inflammation. Furthermore, in vitro, S. miltiorrhiza extracts inhibited p38 and NFκB signaling in Kupffer cells. This effect could be a main mechanism by which S. miltiorrhiza protects against acute liver toxicity.

Discussion and conclusion: Active compounds of S. miltiorrhiza protected the liver from CCl₄-induced injury. Protection might have been due to inhibition of p38 and NFκB signaling in Kupffer cells, which subsequently reduced inflammation in the liver.     

参附对大鼠肾缺血再灌注损伤NF-κB、细胞黏附分子-1、TNF-α表达的影响

目的:观察参附注射液(shenfu injection,SF)对大鼠肾缺血再灌注损伤细胞黏附分子-1(ICAM-1)、NF-κB、TNF-α表达的影响。方法:采用切除右肾,无创动脉夹夹闭左肾动脉45 min,再灌注2 h制作在体肾缺血再灌注损伤模型。36只SD大鼠随机分为缺血再灌注组、假手术对照组、参附预处理组(10 mL/kg),每组12只。免疫组化法检测肾组织中NF-κB、ICAM-1蛋白含量,酶联免疫吸附实验(ELISA)检测血清、肾组织中TNF-α的表达。结果:缺血再灌注组NF-κB、ICAM-1表达和TNF-α含量明显高于假手术对照组(P<0.01)。与缺血再灌注组相比,参附预处理组NF-κB、ICAM-1表达和TNF-α含量明显降低(P<0.01)。结论:参附通过抑制NF-κB的表达,从而下调其下游的TNF-α、ICAM-1的表达,发挥其肾脏保护作用。     

Anti-neuroinflammatory effects of cudraflavanone A isolated from the chloroform fraction of Cudrania tricuspidata root bark

Context: Cudrania tricuspidata Bureau (Moraceae) is an important source of traditional Korean and Chinese medicines used to treat neuritis and inflammation.

Objective: The anti-neuroinflammatory effects of cudraflavanone A isolated from a chloroform fraction of C. tricuspidata were investigated in LPS-induced BV2 cells.

Materials and methods: Cudraflavanone A was isolated from the root of C. tricuspidata, and its structure was determined by MS and NMR data. Cytotoxicity of the compound was examined by MTT assay, indicating no cytotoxicity at 5–40?μM of cudraflavanone A. NO concentration was measured by the Griess reaction, and the levels of PGE₂, cytokines and COX-2 enzyme activity were measured by each ELISA kit. The mRNA levels of cytokines were analysed by quantitative-PCR. The expression of iNOS, COX-2, HO-1, NF-κB, MAPKs and Nrf2 was detected by Western blot.

Results: Cudraflavanone A had no major effect on cell viability at 40?μM indicating 91.5% viability. It reduced the production of NO (IC₅₀?=?22.2?μM), PGE₂ (IC₅₀?=?20.6?μM), IL-1β (IC₅₀?=?24.7?μM) and TNF-α (IC₅₀?=?33.0?μM) in LPS-stimulated BV2 cells. It also suppressed iNOS protein, IL-1β and TNF-α mRNA expression. These effects were associated with the inactivation of NF-κB, JNK and p38 MAPK pathways. This compound mediated its anti-neuroinflammatory effects by inducing HO-1 protein expression via increased nuclear translocation of Nrf2.

Discussion and conclusions: The present study suggests a potent effect of cudraflavanone A to prevent neuroinflammatory diseases. Further investigation is necessary to elucidate specific molecular mechanism of cudraflavanone A.     

姜黄素通过阻断NF-κB减少IL-1β诱导的内皮脂酶表达

目的观察姜黄素对培养的人血管内皮细胞内皮脂酶表达的影响,并探讨其可能的作用机制。方法不同浓度的姜黄素处理HUVEC-12细胞。RT-PCR检测内皮酯酶(endo-thelial lipase,EL)mRNA的表达;Western blot检测核因子-κB抑制因子-α(inhibitor of nuclear factor-κB-α,IκB-α)蛋白的表达;间接免疫荧光检测核因子-κB(nuclear factor-κB,NF-κB)蛋白的活化。结果IL-1β处理HUVEC-12细胞可以明显上调EL mRNA的表达,同时降低胞质蛋白IκB-α的表达水平,激活核转录因子NF-κB,增加胞核蛋白NF-κB的水平。姜黄素预处理HUVEC-12细胞可以抑制IL-1β对EL的上调作用,同时逆转IL-1β诱导的IκB-α蛋白降解和NF-κB活化。结论姜黄素可以通过阻断NF-κB活化减少IL-1β诱导的人血管内皮细胞EL的表达。     

磷脂酰肌醇3-激酶调控白介素-18诱导核因子-кB活化(英文)

目的:研究磷脂酰肌醇(PI)3-激酶在白介素18(IL-18)诱导核因子-кB(NF-кB)活化中的作用。方法:Lipofectin介导反义PI 3-激酶寡核苷酸转染HepG2细胞。用逆转录PCR法检测PI 3-激酶mRNA表达水平,以Sandwich ELISA法检测NF-кB的活化。结果:(1)反义PI 3-激酶寡核苷酸抑制PI 3-激酶mRNA表达。(2)IL-18诱导NF-кB活化。(3)反义PI3-激酶寡核苷酸呈时间(5-24h)和浓度(1-8mg/L)依赖性地抑制IL-18诱导的NF-кB活化。结论:PI 3-激酶调控白介素-18诱导的NF-кB活化。     

羟乙葛根素对大鼠脑缺血再灌注损伤后TNF-α表达及NF-κB活性的影响

观察羟乙葛根素对大鼠局灶性脑缺血再灌注损伤后TNF-α表达及NF-κB活性的影响。采用大鼠大脑中动脉内栓线阻断法(MCAO)建立大鼠脑缺血再灌注损伤模型，分别于缺血前30 min及再灌注即刻由尾静脉注射羟乙葛根素(10，20及40 mg·kg^-1)，缺血2 h再灌注24 h后取缺血侧脑组织，HE染色观察大鼠脑组织病理学变化并计数海马CA1区存活神经元数目，放射免疫分析测定脑组织匀浆中TNF-α含量，逆转录聚合酶链式反应(RT-PCR)测定脑组织中TNF-α mRNA表达情况，凝胶电泳迁移率实验(EMSA)观察NF-κB DNA结合活性改变，Western blotting检测观察IκBα蛋白表达情况。羟乙葛根素可明显改善大鼠海马CA1区损伤程度，升高锥体存活神经元数目，减少TNF-α蛋白及mRNA表达，抑制NF-κB DNA结合活性。羟乙葛根素可减轻大鼠脑缺血再灌注损伤后炎症反应，这可能是其发挥脑保护作用的机制之一。     

急性脊髓损伤时核因子κB与胶质原纤维酸性蛋白相关性研究

目的　明确急性创伤性脊髓损伤 (acutespinalcordinjury,ASCI)后胶质原酸性蛋白 (glialfibril laryacidicprotein ,GFAP)变化分布 ,以及与核因子κB(NF κB)变化的关系。方法　将 2 8只SD大鼠随机分为对照组与损伤组 ,制作脊髓损伤模型 ,使用免疫组化分别测试GFAP与NF κB在急性脊髓损伤后的变化。结果　ASCI后 6h ,白质内GFAP表达开始增强 ,ASCI后 5d ,灰白质内仍有大量的GFAP阳性表现 ;AS CI后 1h ,NF κB的免疫反应在病灶内和边缘部位可以探测到 ;ASCI后 5d ,NF κB的反应明显减少。结论　NF κB部分参与了脊髓损伤后GFAP表达 ,抑制NF κB的表达 ,将一定程度上抑制GFAP表达。