X射线照射对RAW264·7细胞TNF-α和NF-κB表达的影响

目的探讨X射线照射诱发小鼠巨噬细胞分泌TNF-α和NF-κB的规律。方法以8Gy X射线单次照射小鼠巨噬细胞系RAW264.7细胞,并于照后不同时间收集细胞培养上清液,采用ELISA法检测RAW264.7细胞TNF-α分泌水平;以8Gy X射线单次照射小鼠巨噬细胞系RAW264.7细胞,并于照后不同时间收集细胞并进行裂解,采用Western blot实验检测NF-кB（p65）的表达。结果 X射线照射能使小鼠巨噬细胞分泌TNF-α增加,在照射后34h达到一次分泌峰值;细胞核中NF-κB（p65）分布增多,并且随着时间的延长其核移位明显增多,照射后4h达到一次最高值。结论 X射线照射能诱发小鼠巨噬细胞TNF-α和NF-κB表达增强。     

Dimethoxycurcumin, a metabolically stable analogue of curcumin, exhibits anti-inflammatory activities in murine and human lymphocytes

The aim of this study was to investigate whether dimethoxycurcumin (DiMC), a synthetic curcumin analogue having higher metabolic stability over curcumin, could exhibit anti-inflammatory activity in murine and human lymphocytes. Both curcumin and DiMC suppressed mitogen as well as antigen driven proliferation of murine splenic lymphocytes. Further, mitogen and antigen-stimulated cytokine (IL-2, IL-4, IL-6 and IFN-γ) secretion by T cells was also abrogated by curcumin and DiMC. Interestingly, curcumin and DiMC suppressed B cell proliferation induced by lipopolysaccharide. Curcumin and DiMC also inhibited Con A-induced activation of early and late T cell activation markers. They scavenged basal reactive oxygen species and depleted GSH levels in lymphocytes. The suppression of mitogen-induced T cell proliferation and cytokine secretion by curcumin and DiMC was significantly abrogated by thiol containing antioxidants suggesting a role for redox in their anti-inflammatory activity. Further, the possibility of curcumin and DiMC directly interacting with thiol-containing antioxidant GSH was monitored by changes in absorbance. Both curcumin and DiMC inhibited Con A induced activation of NF-κB and MAPK. More importantly, curcumin and DiMC inhibited phytohaemagglutinin induced proliferation and cytokine secretion by human peripheral blood mononuclear cells. To explore their therapeutic efficacy, they were added to lymphocytes post-Con A stimulation and we observed a significant suppression of IL-2, IL-6 and IFN-γ. The present study for the first time demonstrates the potent anti-inflammatory activity of DiMC. Further, DiMC could find application as an alternative to curcumin, which is currently used in several clinical studies, due to its superior bioavailability and comparable efficacy.     

七氟烷后处理对大鼠心肌缺血再灌注NF-κB和TNF-α的影响

目的研究七氟烷后处理对大鼠心肌缺血再灌注过程中NF-κB及TNF-α的影响。方法 40只Wistar雄性大鼠随机分为5组(n=8):假手术组(C组)、缺血再灌注组(I/R组)、1.0%、2.0%、3.0%七氟烷后处理组(S1、S2、S3组)。建立心肌缺血再灌注模型。光镜下观察心肌组织病理学变化,免疫组化测大鼠NF-κB的活性,ELISA法测TNF-α的浓度。结果与C组相比,I/R组、各S组NF-κB的活性及TNF-α的浓度显著升高(P<0.05);与I/R组相比,各S组NF-κB的活性及TNF-α的浓度显著降低(P<0.05);S1组与S3组各指标间无明显差异(P>0.05);与S1组、S3组相比,S2组NF-κB的活性及TNF-α的浓度显著降低(P<0.05)。结论七氟烷后处理可通过抑制NF-κB的活性,降低TNF-α的表达水平,达到心肌保护作用,且浓度为2.0%的七氟烷效果最佳。     

蛛网膜下腔出血后迟发性脑血管痉挛中 TNF-α和NF-кB 的表达变化

目的：检测蛛网膜下腔出血（以下简称 SAH）后迟发性脑血管痉挛中TNF-α和NF-кB的表达变化。方法56只中国白兔随机分成3组：对照组（ n ＝8）：不做枕大池穿刺注血,处死;SAH组（ n ＝24）：在第0天和第2天分二次枕大池注射自体动脉血,分别在第1次注血后第4天、第7天和第14天分三批处死,每组8只;假手术组（ n＝24）：只做枕大池穿刺不注血,第4天、第7天和第14天处死,每组8只。通过计算动脉口径比判断基底动脉痉挛程度,使用免疫组化和免疫蛋白印迹的方法检测迟发性脑血管痉挛中TNF-α和NF-кB的表达变化。结果枕大池注血方法可以造成SAH后迟发性脑血管痉挛模型。 NF-κB 和 TNF-α在注血后第4天表达开始增加（ P ＜0．01）,在第7天达到最高值（ P ＜0．01）,第14天降到对照组水平。结论蛛网膜下腔出血后迟发性脑血管痉挛中NF-κB及 TNF-α表达增加,炎症反应在此过程中起着重要作用。     

SN50对缺血再灌注损伤中TNF-α影响的实验研究

目的研究核转录因子-κB（nuclear factor-kappaB,NF-κB）抑制剂SN50对氧糖剥夺条件下培养的SD大鼠巨噬细胞炎性因子肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）表达的影响。方法用MTT法检测SN50对细胞增殖活性的影响,以ELISA法检测氧糖剥夺条件下SN50对TNF-α表达影响。结果经SN50处理的氧糖剥夺条件下的巨噬细胞成活率较未经处理的明显上升（P〈0.05或P〈0.01）,TNF-α表达的上调幅度亦明显变小（P〈0.05或P〈0.01）。结论 SN50主要是通过抑制NF-κB核的移位干扰了TNF-α的基因转录而导致其合成的蛋白量降低。     

柚皮苷对人皮肤角质细胞分泌趋化因子RANTES及其核转录因子-kB信号通路影响的实验研究

目的：研究炎症性细胞因子肿瘤坏死因子α（TNF-α）和γ干扰素（IFN-γ）对人皮肤角质细胞（HaCaT）分泌趋化因子RANTES的诱导作用;以阿维A为阳性对照药,研究柚皮苷对此诱导作用的抑制作用及其机制。方法：将HacaT分为阴性对照组、模型刺激组（TNF-α＋IFN—γ）、阿维A预处理组（TNF-α＋IFN—γ＋阿维A）和柚皮苷预处理组（TNF-α＋IFN—γ＋柚皮苷）等;采用酶联免疫吸附（ELISA）法测定HaCaT细胞经单用和合用TNF-α、IFN-γ刺激以及加入阿维A和柚皮苷后RANTES的分泌量。采用免疫细胞化学方法和Western blot法检测HaCaT中核转录因子-KB P65（NF—kB P65）蛋白的表达。结果：单用或合用TNF—α、IFN-γ可显著诱导细胞分泌RANTES,并以合用组作用最显著（P〈0．01）。柚皮苷、阿维A均能明显抑制TNF-α、IFN-γ诱导的RANTES的分泌（P〈O．01）及NF—kB P65蛋白的表达。结论：柚皮苷可以抑制TNF-α、INF-γ诱导RANTES的分泌及相关蛋白的产生,其机制可能是通过抑制NF—kB信号传导途径的活化而实现。     

金纳多注射液对脂多糖致急性肺损伤大鼠肺组织中核因子-κB表达的影响

目的：探讨金纳多注射液对脂多糖致急性肺损伤（ ALI）大鼠肺组织中核因子-κB（ NF-κB）表达的影响。方法采用随机法将24只Wistar大鼠分为3组：对照组（ Control）、脂多糖组（ LPS）和金纳多处理组（ GBE）。采用尾静脉注射脂多糖（ LPS）的方法建立急性肺损伤模型,光学显微镜下观察每组大鼠的肺组织病理学改变;检测血清中肿瘤坏死因子-αB（ TNF-αB）和肺组织中核因子-κB（NF-κB）的表达变化。结果与对照组相比,脂多糖组大鼠血清中TNF-αB含量升高（P＜0．05）,肺组织中NF-κB表达增加（P＜0．05）;与脂多糖组相比,金纳多处理组大鼠血清中TNF-αB含量减少（P＜0．05）,肺组织中NF-κB表达减少（P＜0．05）。结论金纳多注射液可有效减轻ALI时肺组织的炎症反应,其机制可能与其降低大鼠体内TNF-α含量、减少NF-κB表达有关。     

白及多糖对溃疡性结肠炎的作用研究

目的:研究白及多糖(BSPS)对溃疡性结肠炎(UC)的作用。方法:小鼠均分为6组,即对照(等容生理盐水)、模型(等容生理盐水)、柳氮磺胺吡啶(SASP,370mg.kg-1)和BSPS高、中、低剂量(320、160、80mg.kg-1)组,ig给药,每天1次,连续10d。恶唑酮诱导复制UC模型后观察小鼠疾病活动指数(DAI)、结肠指数、结肠组织大体评分和病理评分,并检测结肠组织白细胞介素(IL)-10、肿瘤坏死因子(TNF)-α和核因子(NF)-κB的水平。结果:与模型组比较,BSPS高、中剂量组小鼠DAI、结肠指数、结肠组织大体评分、病理评分和TNF-α、NF-κB水平均显著降低(P<0.05或P<0.01),IL-10显著升高(P<0.05)。结论:BSPS具有促进UC模型小鼠肠黏膜修复、抑制机体炎症和恢复免疫平衡的作用,其作用机制可能与抑制TNF-α和NF-κB,上调IL-10水平有关。     

Immune response mechanism of mouse monocytes/macrophages treated with κ-carrageenan polysaccharide

This study investigated the function of κ-carrageenan polysaccharide in immune regulation. The immune response of RAW 264.7 cells treated with κ-carrageenan polysaccharide was explored by MTT assay, general morphological observation, neutral red phagocytosis assay, Griess method, fluorescence method, and enzyme-linked immunosorbent assay. In addition, TLR4 blocking experiment and double-fluorescence immunostaining were performed on cells to demonstrate their immune response mechanism. Results show that κ-carrageenan polysaccharide not only promotes cell proliferation but also activates RAW 264.7 cells, thereby improving the cells’ phagocytic capability, NO production, and tumor necrosis factor-α (TNF-α) secretion. In addition, the use of TLR4-specific inhibitors can significantly mediate the increased TNF-α secretion induced by κ-carrageenan polysaccharide. The RAW 264.7 cells treated with κ-carrageenan polysaccharide show upregulated TLR4 expression, and the main subunit of NF-κB (p65) is translocated. These results support the immunomodulatory function of κ-carrageenan polysaccharide in RAW 264.7 cells.     

Okra polysaccharide-2 plays a vital role on the activation of RAW264.7 cells by TLR2/4-mediated signal transduction pathways

Polysaccharide is the main active component of okra (Abelmoschus esculentus L.) and it can effectively stimulate the activation of macrophages. However, the immune regulatory mechanism is still not clear. Therefore, the present study aimed to reveal the possible mechanism by investigating the effect of okra polysaccharide-2 (RPS-2) on Toll-like receptor (TLR) 2/4-mediated signal transduction pathways in RAW264.7 murine macrophage cells. In order to confirm whether RPS-2 stimulated macrophages activation via TLR2 or TLR4, RAW264.7 murine macrophage cells were pretreated with TLR2/4 inhibitors for 1 h before RPS-2 treatment, and then the NO, IL-10, TNF-α levels were tested. The results indicated that both TLR2 and TLR4 were the keys of immune regulatory effect of RPS-2. Afterwards, the effect of RPS-2 on NF-κB and MAPKs signaling pathways were studied by western blot analysis. It showed RPS-2 induced the phosphorylation of p65, IκBα, p38, ERK1/2 and JNK. At the same time, the specific inhibitors reduced these phosphorylation levels as well as NO, IL-10 and TNF-α amounts. In a word, RPS-2 activated macrophages by NF-κB and MAPKs signal transduction pathways.     

Plumbagin inhibits LPS-induced inflammation through the inactivation of the nuclear factor-kappa B and mitogen activated protein kinase signaling pathways in RAW 264.7 cells

Plumbagin (PL) has been reported to exhibit anti-carcinogenic, anti-inflammatory and analgesic activities, but little is known about its mechanism. In this study, we investigated the anti-inflammatory property of PL and its mechanism of action. Although no significant cytotoxicity of PL was observed over the concentration range tested, PL (2.5–7.5 μM) significantly and dose-dependently suppressed the secretion of pro-inflammatory mediators and inhibited the expression of TNF-α, IL-1β, IL-6 and iNOS in LPS-stimulated RAW 264.7 cells. Furthermore, PL consistently suppressed the activity of iNOS in LPS-induced RAW 264.7 cells. To elucidate the mechanism underlying the anti-inflammatory activity of PL, we assessed the effects of PL on the MAPK pathway and the activity and expression of NF-κB. These experiments demonstrated that PL significantly reduced the luciferase activity of an NF-κB promoter reporter and p65 nuclear translocation. The LPS-induced phosphorylation of MAP kinases was also attenuated by PL; significant changes were observed in the levels of phosphorylated ERK1/2, JNK and p38 MAPK. Additionally, MAPK inhibitors confirmed the inhibitory effect of PL on the MAPK pathway. Taken together, these data suggest that PL exerts its anti-inflammatory effects by down-regulating the expression of pro-inflammatory mediators through inhibition of NF-κB and MAPK signaling in LPS-stimulated RAW 264.7 cells.     

泻心汤有效组分不同配伍对内毒素肺损伤大鼠NF-κB及IκB表达的影响

目的研究泻心汤有效组分及其配伍对内毒素肺损伤大鼠NF-κB及IκB表达的影响。方法大鼠灌胃给予黄芩总黄酮提取物(TFL)、大黄总游离蒽醌提取物(TFA)、大黄总结合蒽醌提取物(TCA)、TFL与TFA配伍高、低剂量(A高A低)、TFL与TCA配伍(B)及醋酸地塞米松(Dex)4d,末次给药后1h股静脉注射脂多糖(LPS)建立大鼠内毒素肺损伤模型,1、2、4h各组分别处死6只动物,收集肺组织,免疫印记(Western blot)检测核因子κB(NF-κB)、κB抑制因子(IκB)的蛋白表达。结果大黄总游离蒽醌、A高及Dex在1、2、4h均能够明显抑制NF-κB p65的核转位(P<0.01),所有给药组在2、4h均能够明显抑制胞质中的IκBα的降解(P<0.01)。结论泻心汤有效组分及其配伍对内毒素肺损伤大鼠保护作用与抑制NF-κB的核转位及IκB的降解有关。     

柚皮苷对人皮肤角质细胞分泌趋化因子RANTES及其核转录因子-κB信号通路影响的实验研究

目的:研究炎症性细胞因子肿瘤坏死因子α(TNF-α)和γ干扰素(IFN-γ)对人皮肤角质细胞(HaCaT)分泌趋化因子RANTES的诱导作用;以阿维A为阳性对照药,研究柚皮苷对此诱导作用的抑制作用及其机制。方法:将HaCaT分为阴性对照组、模型刺激组(TNF-α+IFN-γ)、阿维A预处理组(TNF-α+IFN-γ+阿维A)和柚皮苷预处理组(TNF-α+IFN-γ+柚皮苷)等;采用酶联免疫吸附(ELISA)法测定HaCaT细胞经单用和合用TNF-α、IFN-γ刺激以及加入阿维A和柚皮苷后RANTES的分泌量。采用免疫细胞化学方法和Western blot法检测HaCaT中核转录因子-κBP6(5NF-κBP65)蛋白的表达。结果:单用或合用TNF-α、IFN-γ可显著诱导细胞分泌RANTES,并以合用组作用最显著(P<0.01)。柚皮苷、阿维A均能明显抑制TNF-α、IFN-γ诱导的RANTES的分泌(P<0.01)及NF-κBP65蛋白的表达。结论:柚皮苷可以抑制TNF-α、INF-γ诱导RANTES的分泌及相关蛋白的产生,其机制可能是通过抑制NF-κB信号传导途径的活化而实现。     

Does omethoate have the potential to cause insulin resistance?

Persistent organic pollutant exposure is strongly associated with the development of diabetes. The development of diabetes or alteration in blood glucose levels is associated with insulin resistance that precedes diabetes for many years. Omethoate is a commonly used insecticide in most developing countries. The present study was designed to elucidate the potent role of omethoate in developing insulin resistance in rats. Male Wistar rats were exposed to omethoate at the concentration of 1.5 mg/kg body weight (1/40 LD₅₀), 3 mg/kg body weight (1/20 LD₅₀) and 6 mg/kg body weight (1/10 LD₅₀) through gastric injection for 60 days; control group rats received PBS through gastric injection. The results showed that the levels of MDA, TNF-α and IL-6 were increased and the activities of SOD and GSH-Px were decreased in the right thigh muscles of rats exposed to omethoate. However, JNK, p38 MAPK and NF-κB in right thigh muscles of rats exposed to omethoate were activated. This study suggested that omethoate had a potential to cause insulin resistance.     

丹参注射液预防大鼠急性胰腺炎及其机制研究

目的观察丹参注射液预防大鼠急性胰腺炎及其对血清肿瘤坏死因子(TNF-α)、白细胞介素(IL-6)、胰腺组织核因子(NF-κB)及氧自由基的影响。方法将60只SD大鼠随机分为6组,分别为假手术组、模型组、乌司他丁组(20 000U/kg)以及丹参注射液低、中、高剂量(0.4、0.8、1.6 m L/kg)组。用5%牛磺胆酸钠(1.0 m L/kg)胰管内注入诱导大鼠急性胰腺炎模型,造模前30 min分别尾iv给予丹参注射液。各组大鼠均于造模6 h后行腹主动脉采血,处死大鼠取胰腺组织标本,检测SD大鼠急性胰腺炎模型血清淀粉酶(AMY)、血浆内毒素(ET)、胰腺病理评分和病死率。并用ELISA法检测血清IL-6、TNF-α浓度的变化,运用免疫组化法检测NF-κB表达水平,同时切取各时间点大鼠胰腺组织,检测丙二醛(MDA)、组织髓过氧化物酶(MPO)及超氧化物歧化酶(SOD)的水平。结果模型组大鼠的血清AMY、血浆ET、病理评分、死亡率、IL-6、TNF-α水平及胰腺组织中NF-κB活性、MDA水平和MPO活性均较假手术组升高(P0.01),而SOD活性则明显降低(P0.05);乌司他丁组和丹参注射液组上述各项指标均较模型组明显改善(P0.05、0.01)。结论丹参注射液可显著降低实验性急性胰腺炎大鼠的IL-6、TNF-α及NF-κB,清除氧自由基,减轻脂质过氧化反应,从而减轻急性胰腺炎病理损害程度,降低急性胰腺炎大鼠的死亡率。     

Suppression of endotoxin-induced proinflammatory responses by citrus pectin through blocking LPS signaling pathways

Pectin is composed of complex polysaccharides rich in galactoside residues, and it is most abundant in citrus fruits. Pectin and modified pectin have been found to exhibit anti-mutagenic activity and inhibit cancer metastasis and proliferation, with no evidence of toxicity or other serious side effects. In this study, we investigated the inhibitory effect of pectin at different degrees of esterification (DEs) on the expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-activated macrophages. Western blot and RT-PCR analyses demonstrated that 30% esterified pectin (DE30), DE60 pectin, and DE90 pectin significantly inhibited the protein and mRNA expressions of iNOS and COX-2 in LPS-activated macrophages, and DE90 pectin was the most-potent inhibitor. To clarify the mechanisms involved, DE90 pectin was found to inhibit the phosphorylation of MAPKs and IKK kinase activity. In addition, DE90 pectin inhibited the activation of NF-kappaB and AP-1 by electrophoretic mobility shift assay and transient transfection experiments. Finally, we found that DE90 pectin could bind with LPS, and might result in decreased binding of LPS to its receptor. These results suggest that modulation of iNOS and COX-2 expressions by dietary pectin may be important in cancer chemoprevention and anti-inflammation.     

基于HPLC-Q-Orbitrap-MS槐白皮水提物的化学成分测定及其对溃疡性结肠炎的治疗作用研究

目的 分析槐白皮水提物的化学成分,探讨其对溃疡性结肠炎的治疗作用及机制.方法 采用高效液相色谱-四级杆/静电场轨道阱高分辨质谱(HPLC-Q-Orbitrap-MS)技术对槐白皮水提物的化学成分进行定性鉴定,与数据库(mzCloud、mzVault、ChemSpider)检索比对.42只♂ BALB/c小鼠随机分为正常...