核因子κB抑制蛋白家族的研究进展

核因子κB抑制蛋白(inhibitor ofNF-κB,IκB)是核因子κB(nuclear factor-kappa B,NF-κB)的抑制因子。IκB蛋白家族与Rel/NF-κB蛋白家族一样,也是一个大家族,它们都来源于相同祖先NF-κB/IκB超家族的Rel同源区-锚蛋白重复序列区(Rel homology domain-ankyrin repeat domain,RHD-ARD)结构[1]。IκB蛋白一方面可以在胞浆中与NF-κB二聚体结合形成复合物,抑制NF-κB进入细胞核发挥转录激活作用;另一方面也可以直接在细胞核内抑制NF-κB与DNA的结合。IκB蛋白家族的成员主要包括:IκBα、IκBβ、IκBε、Bcl-3、IκB…     

腺病毒介导核因子κBp65特异性小干涉RNA抑制骨关节炎

背景：前期系列研究表明,核因子κBp65特异性小干涉RNA重组腺病毒可以在SD大鼠膝关节软骨和滑膜内抑制核因子κBp65的表达,降低核因子κB的转录活性,抑制关节内白细胞介素1β和肿瘤坏死因子α的表达。 目的：在大鼠关节内观察腺病毒介导的核因子κBp65特异性小干涉RNA对骨关节炎的抑制作用。 方法：将3月龄雄性近交系SD大鼠分为4组,正常关节组只切开皮肤及关节腔后立即缝合切口。其余大鼠切断膝关节内侧副韧带,切除部分内侧半月板,诱导大鼠关节的骨关节炎。骨关节炎组造模后未作任何处理;空病毒注射组造模后两侧膝关节关节腔注射腺病毒空载体0.2 mL;特异性小干涉RNA注射组造模后两侧膝关节关节腔注射携带核因子κBp65特异性小干涉RNA重组腺病毒0.2 mL。 结果与结论：与正常关节软骨相比,骨关节炎组和空病毒注射组关节软骨组织评分及滑膜组织评分显著增高(P < 0.01),特异性小干涉RNA注射组组织评分明显减低(P < 0.01),但未到正常关节软骨和滑膜水平(P < 0.01)。提示腺病毒介导的核因子κBp65特异性小干涉RNA可以抑制骨关节炎的发展。     

烧伤血清对单核细胞抑制性κB降解和核因子-κB活化的影响

目的:了解烧伤血清对单核细胞抑制性κB(IκBα)降解、核因子-κB(NF-κB)活化的影响,进一步探讨烧伤血清诱导单核细胞活化分泌细胞因子的机理。方法:采用体外培养的人外周血单核细胞(PBMC),分别用正常人血清(对照组)、烧伤患者血清、烧伤患者血清+吡咯烷二硫代氨基甲酸盐(PDTC)刺激单核细胞,采用Western印迹法检测血清刺激30、60、90、120min后单核细胞IκBα蛋白降解情况,电泳迁移率分析检测血清刺激30、60、120、240min后NF-κB活性的变化。结果:烧伤血清刺激单核细胞后30min,IκBα发生明显降解,刺激60min达高峰,2h后表达逐渐升高。而烧伤血清刺激单核细胞后30min,NF-κB活性迅速升高,30-60min达高峰,2h后接近基础状态。PDTC能有效抑制烧伤血清作用条件下单核细胞IκBα降解、NF-κB活化。结论:烧伤血清可诱导单核细胞IκBα降解,活化NF-κB,从而在机体烧伤后单核细胞分泌细胞因子过程中起重要作用。     

姜黄素保护关节软骨抑制骨关节炎的作用和机制

背景：研究证实姜黄素具有抗炎抗氧化抗凋亡作用。 目的：综合阐述姜黄素保护关节软骨及抑制骨关节炎等方面的机制和作用。 方法：作者检索 1973至2014 年 PubMed、Embase、Elseveir数据库文献。检索词为“Osteoarthritis,curcumin,chondrocyte,articular cartilage”,按照事先制定的标准逐一评价纳入研究的文献,提取有效资料进行综合分析。 结果与结论：姜黄素可通过抑制氧化作用酶,清除自由基,完成抗氧化作用,从而防止骨关节炎的发生和进展。姜黄素抑制基质金属蛋白酶的对软骨基质的消耗,增加Ⅱ型胶原的产生。姜黄素通过抑制胞浆型磷脂酶A2,环氧化酶2,脂氧合酶5,从而完成抗炎反应。姜黄素抑制白细胞介素1β导致的线粒体肿胀和凋亡,完成抗凋亡作用。临床研究提示,姜黄素或者其衍生物骨关节炎患者膝关节功能、关节疼痛等均有改善。高浓度姜黄素对体外培养的细胞和组织有毒性作用。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程全文链接：     

硝普钠对核因子κB活性影响的初步研究

目的:探讨不同浓度的硝普钠(SNP)对人外周血T淋巴细胞核因子kappa B(NF-κB)活性的影响及机制。方法: 应用人外周血T淋巴细胞培养、Western blot和RT-PCR等技术,观察不同浓度的一氧化氮(NO)供体SNP对人T细胞受植物血凝素(PHA-P)刺激30 min、120 min后IκB_α mRNA和蛋白质表达的影响。结果:中高浓度的SNP能显著减少PHA-P刺激30 min时IκB_α蛋白的降解,并明显增加刺激120 min时IκB_α mRNA的再表达。结论: 中高浓度的SNP抑制NF-κB活性的机制与其通过NO减少IκB_α降解、促进IκB_α再合成有关,而低浓度SNP对NF-κB活性影响的机制可能不是通过IκB_α。     

基质细胞衍生因子1对骨关节炎软骨Ⅱ型胶原及分泌基质金属蛋白酶的影响*

背景：基质细胞衍生因子1/趋化因子受体4信号通路在骨关节炎的发病中起关键作用。 目的：观察外源性基质细胞衍生因子1对骨关节炎软骨Ⅱ型胶原及分泌基质金属蛋白酶的影响。 方法：分别取因骨关节炎行膝关节置换患者软骨组织块40块,作为实验组;取因创伤性截肢患者膝关节的正常软骨块40块,作为对照组;置入DMEM培养液中,加入79,392 μg/L外源性基质细胞衍生因子1,分别培养48 h和96 h。 结果与结论：实验组与对照组培养48,96 h后,各时间段Ⅱ型胶原免疫组织化学染色皆呈阳性,但高浓度基质细胞衍生因子1培养液条件下时间越长Ⅱ型胶原降解越严重;实验组与对照组同一时间段同浓度基质细胞衍生因子1培养液条件下基质细胞衍生因子1水平无明显差异(P > 0.05);高浓度基质细胞衍生因子1培养液条件下同一时间段实验组比对照组中促使软骨释放基质金属蛋白酶多(P < 0.05)。提示基质细胞衍生因子1可通过基质细胞衍生因子1/趋化因子受体4信号通路调节骨关节炎软骨基质金属蛋白酶表达并致软骨Ⅱ型胶原降解。     

骨髓间充质干细胞经核转录因子κB途径干预心肌纤维化

背景：骨髓间充质干细胞移植能否直接干预心肌纤维化及其可能的机制尚不完全清楚。 目的：观察骨髓间充质干细胞移植干预心肌纤维化的效果并分析其机制。 方法：分离培养雄性小鼠骨髓间充质干细胞,经尾静脉输入异丙肾性心肌纤维化雌性小鼠为治疗组,另设未治疗组和正常对照组。5周后处死小鼠,实时荧光定量PCR检测心肌y染色体鉴别基因(SRY)、基质金属蛋白酶9、基质金属蛋白酶组织抑制剂1的表达;天狼猩红染色对比心脏胶原纤维含量;免疫组织化学染色法观察心脏核转录因子κB表达。 结果与结论：骨髓间充质干细胞能归巢于纤维化心肌。与未治疗组相比,治疗组的心肌基质金属蛋白酶9、基质金属蛋白酶组织抑制剂1和核转录因子κB表达下调(P < 0.05),胶原纤维含量下降(P < 0.05)。结果表明,骨髓间充质干细胞移植干预心肌纤维化,其机制可能与抑制核转录因子κB过度活化有关。     

关节腔内注射透明质酸钠可以延缓创伤性骨关节炎的软骨退变

背景：透明质酸钠是一种治疗骨关节炎的有效手段,但其机制尚不清楚。有研究表明基质金属蛋白酶1,3,9和基质金属蛋白酶组织抑制剂1,2表达失调对骨关节炎有重要影响。目的：观察关节腔内注射透明质酸钠对兔创伤性骨关节炎模型软骨中基质金属蛋白酶1,3,9和基质金属蛋白酶组织抑制剂1,2表达的影响。方法：采用单侧前交叉韧带切断法构建创伤性骨关节炎模型兔,造模后4周关节腔注射体积分数1%透明质酸钠0.3 mL,每周1次,连续5周,设为透明质酸钠组,同时设模型组和正常对照组作对比。术后11周麻醉处死动物,获取软骨并提取总RNA,用实时聚合酶链反应分析各组软骨内基质金属蛋白酶1,3,9和基质金属蛋白酶组织抑制剂1,2 mRNA表达。结果与结论：与模型组相比,透明质酸钠组经关节腔内注射透明质酸钠软骨损伤范围和程度均减轻(P < 0.01),软骨组织学评分明显降低(P < 0.05)。模型组软骨内基质金属蛋白酶1,3,9 mRNA表达增强,基质金属蛋白酶组织抑制剂1,2 mRNA表达下调,而透明质酸钠组软骨中基质金属蛋白酶1,3,9,基质金属蛋白酶组织抑制剂1,2 mRNA的表达并无显著变化。结果说明,基质金属蛋白酶1,3,9和基质金属蛋白酶组织抑制剂1,2参与了创伤性骨关节炎软骨退变过程,虽然关节腔内注射透明质酸钠可以延缓创伤性骨关节炎软骨退变,但透明质酸钠并不是通过调节上述因子的表达来发挥修复作用的,具体机制有待进一步研究证实。中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

实验性根尖周炎模型大鼠核因子?B的表达

BACKGROUND:mRNA expression level of nuclear factor-kappa B (NF-?B) in periapical periodontitis has been shown to be significantly higher than that in normal periapical tissue. OBJECTIVE:To observe the expression of NF-?B in rats with periapical periodontitis. METHODS:Twenty male Sprague-Dawley rats aged 6 weeks were enrolled to establish apical periodontitis model at the right mandibular first motar (experimental group), and the left mandibular first motar as normal control group. Five rats were randomly selected at postoperative 1, 2, 3, and 4 weeks to make frozen sections of the mandibular tissue. The morphology of the periapical tissue was observed through hematoxylin-eosin staining, and the expression level and distribution of NF-?B were detected by immunohistochemical staining. RESULTS AND CONCLUSION: (1) Hematoxylin-eosin staining showed that the apical periodontal tissue of the mandibular first molar at the right side had local inflammatory response and spread to peripheral tissues; while there was no pathological changes in the control group. (2) Immunohistochemistry results revealed that NF-?B expression increased from the 1st week to 2nd week after surgery, and then went down at the 3rd week and reached the lowest point at the 4th week; moreover, the expression level at each time exhibited significant differences (P < 0.05). The amount of NF-?B-positive cells in the experimental group was significantly higher than that in the control group (P < 0.05). (3) These findings suggest that the expression level of NF-?B holds close correlation with the process of periapical periodontitis indicating that NF-?B may participate in the inflammatory reaction of periapical periodontitis.  中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程     

动脉粥样硬化模型大鼠主动脉Rho激酶及相关细胞因子表达与益气活血方的干预

BACKGROUND: Representative formulae of Qi-Benefiting and Blood-Activating Chinese medicinal herbs Bu Yang Huan Wu Tang (BYHWT) can improve microcirculation and hypoxia, decrease fibrinogen, resist platelet aggregation, reduce blood lipid, dilate blood vessel, resist thrombosis, and reduce neonatal plaque rupture, and has a multi-target, multi-channel anti-atherosclerotic effect.      

富血小板血浆修复膝关节骨关节炎

BACKGROUND: Platelet-rich plasma (PRP) containing high levels of platelet-derived growth factor for knee osteoarthritis has achieved good clinical results; however, the effects of RPR on the repair of damaged articular cartilage are still controversial. OBJECTIVE: To study the effects of RPR on the repair of damaged articular cartilage in a rabbit model of knee osteoarthritis. METHODS:The model of osteoarthritis in the rabbit right knee was established by Hulth’s method. Autologous PRP (0.5 mL) (PRP group), sodium hyaluronate (0.5 mL) (sodium hyaluronate group), and normal saline (model group) were injected into the right knee joint cavity, respectively. The morphology of articular surface and nitric oxide contents in knee joint fluid were observed and determined at 8 weeks after surgery. RESULTS AND CONCLUSION: The morphology of articular cartilage in the PRP group was better than that in the other three groups. Mankin scores of articular cartilage and nitric oxide contents of knee joint fluid in the PRP group were significantly decreased compared with the model and sodium hyaluronate groups that in (P < 0.05), while increased compared with the sham-operation group (P < 0.05). Our results suggest that repair effects of PRP on the damaged articular cartilage are superior to sodium hyaluronate treatment. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

慢病毒介导shRNA关节腔注射可加重骨关节炎的软骨破坏

BACKGROUND: Emerging evidence demonstrates that low density lipoprotein receptor-related protein 1 (LRP1) is involved in lipid metabolism and regulation of inflammatory reaction. OBJECTIVE:To explore the effect of lentivirus-induced knockdown of low density lipoprotein receptor-related protein 1 on cartilage damage and matrix metalloproteinase 13 in a rat model of osteoarthritis, so as to assess the role of low density lipoprotein receptor-related protein 1 in the pathogenesis of osteoarthritis. METHODS: Sixty-four Sprague-Dawley rats were included and ramdomly divided into four groups (n=16 for each): negative control group, no surgery; sham-surgery group, only the articular cavity of the knee was exposed; osteoarthritis plus shLRP1 group, rat osteoarthritis models were established by cutting anterior cruciate ligament and removing the medial meniscus partly followed by an intra-articular injection of lentivirus-mediated siRNA at 2 days after surgery, once a week for 2 consecutive weeks; osteoarthritis group, an intra-articular injection of the negative control lentivirus was performed after surgery. Rats in the four groups started running on the self-made electric treadmill from 5 days after modeling, 30 minutes per day, totally 500 meters. Cartilage damage and matrix metalloproteinase 13 expression in cartilage tissues were determined at 2, 4, 6 weeks after surgery. RESULTS AND CONCLUSION: Gross and pathological observations showed that lentivirus-induced knockdown of low density lipoprotein receptor-related protein 1 aggravated cartilage damage in the rat model of osteoarthritis. At 6 weeks after surgery, Mankin’s score and matrix metalloproteinase 13 expression in the cartilage tissues in osteoarthritis plus shLRP1 group were significantly increased compared with other three groups (P < 0.05). These findings indicate that a simulation model of osteoarthritis is developed by cutting anterior cruciate ligament and removing the medial meniscus partly combined with running on the treadmill. Lentivirus-induced knockdown of LRP1 aggravates cartilage damage in a rat model of osteoarthritis 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

Culture Duration Modulates Collagen Hydrolysate-Induced Tissue Remodeling in Chondrocyte-Seeded Agarose Hydrogels

Media supplementation with collagen hydrolysate was hypothesized to increase the collagen content in engineered cartilage. By d28, hydrolysate at 0.5 mg/mL increased type II collagen content and 1 mg/mL increased mechanical properties, total collagen content, and type II collagen content over controls. By d42, however, controls possessed the highest GAG content and compressive Young’s modulus. Real-time PCR found that 1 mg/mL increased type II collagen gene expression in d0 constructs, but increased MMP expression with no effect on type II collagen on d28. A 10 mg/mL concentration produced the lowest tissue properties, the lowest type II collagen gene expression on d0, and the highest MMP gene expression on d28. These results indicate that the duration of culture modulates the response of chondrocytes to collagen hydrolysate in 3D culture, transforming the response from positive to negative. Therefore, collagen hydrolysate as a media supplement is not a viable long-term method to improve the collagen content of engineered cartilage tissue.     

腰突颗粒对人髓核细胞核因子κB信号通路的影响

BACKGROUND: Nuclear factor-κB (NF-κB) is highly expressed in the degenerated intervertebral disc. The Herbal Compound formula Yaotu Granules for lumbar disc herniation has achieved good clinical results; however, the underlying mechanisms remain poorly understood. OBJECTIVE: To explore mechanisms underlying the effects of the drug serum of Yaotu Granules on NF-κB signaling pathway. METHODS: Eight New Zealand white rabbits were randomly divided into normal saline group, low-, middle-, and high-dose drug groups. Normal saline, 10, 20, and 40 mL of water decoction of Yaotu Granules were administered intragastrically in the normal saline, low-, middle-, and high-dose drug groups for 7 days, respectively. The drug serum of Yaotu Granules was prepared using serum samples obtained from the common carotid artery at 2 hours after the last administration. Human nucleus pulposus cells were maintained in DMEM containing 10% (v/v) drug serum for 24 hours. RESULTS AND CONCLUSION: The expression levels of interleukin (IL)-8, tumor necrosis factor (TNF)-α, p-P65, P50, IκB-α (the NF-κB inhibitor), and IκB kinase β (IKK-β) mRNA in human nucleus pulposus cells in three drug serum groups were significantly decreased compared with the normal saline group; in addition, a similar change in the expression levels of p-P65, P50, IκB-α, and IKK-β protein between groups were detected. Noticeably, the expression levels of IL-8, TNF-α, p-P65, P50, IκB-α, and IKK-β mRNA, as well as the expression levels of p-P65, P50, IκB-α, and IKK-β protein were gradually decreased with the increased drug dose. All these results suggest that the drug serum of Yaotu Granules can protect nucleus pulposus cells and delay their degeneration through blocking NF-κB signaling pathway. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

雾化吸入灭活草分枝杆菌降低哮喘小鼠核因子κB及细胞间黏附分子1的表达

 目的:研究雾化吸入灭活草分枝杆菌对支气管哮喘小鼠气道炎症,以及哮喘肺组织中核因子κB(NF-κB)、细胞间黏附分子1(ICAM-1)和血管细胞黏附分子1(VCAM-1)的影响,探讨雾化吸入灭活草分枝杆菌防治哮喘的机制。方法:将24只雄性BALB/c小鼠按随机数字表法分为3组,每组8只：正常对照组（A）、哮喘模型组（B）和治疗组（C）。以鸡卵清蛋白致敏制造小鼠支气管哮喘模型。C组在激发后给予雾化吸入灭活草分枝杆菌治疗5 d,每天1次。各组动物处死后提取肺组织和支气管肺泡灌洗液（BALF）。进行病理HE染色及AB-PAS染色观察气道炎症浸润及黏液分泌情况,并行病理半定量分析。对BALF中炎症细胞进行分类计数。实时荧光定量PCR检测肺组织NF-κB、ICAM-1和VCAM-1的mRNA表达水平。结果:治疗组嗜酸性粒细胞比例低于模型组(P<0.05),气道炎症病变及黏液分泌情况较模型组减轻(P<0.05, P<0.01)。哮喘模型组的肺组织中NF-κB mRNA含量与正常组相比显著升高(P<0.01),而治疗组肺组织中的NF-κB mRNA 含量明显低于模型组(P<0.05);模型组的ICAM-1 mRNA 水平比正常组高(P<0.05),但治疗后明显降低(P<0.01);VCAM-1的 mRNA水平在各组间无显著差异。相关性检验发现小鼠肺组织中VCAM-1的mRNA与ICAM-1的mRNA呈明显正相关(r=0.84,P<0.01),但NF-κB的mRNA与ICAM-1的mRNA、VCAM-1的mRNA无明显相关性(均P>0.05)。结论:雾化吸入草分枝杆菌对支气管哮喘小鼠气道炎症及黏液分泌有抑制作用;NF-κB参与哮喘发病过程,雾化吸入灭活草分枝杆菌降低哮喘小鼠的NF-κB水平。同时雾化吸入灭活草分枝杆菌可降低黏附分子尤其是ICAM-1的表达,是其控制炎症的另一个重要机制。     

脂肪间充质干细胞移植治疗1型糖尿病模型大鼠的作用

背景:干细胞移植是1型糖尿病比较有前景的治疗方法,脂肪间充质干细胞是继骨髓间充质干细胞后的又一研究热点。目的:观察脂肪间充质干细胞移植对1型糖尿病大鼠的治疗作用。方法:将45只SD大鼠随机分为正常组、模型组和细胞移植组,后2组采用链脲菌素腹腔注射建立1型糖尿病模型。造模后7 d,正常组腹腔静脉注射生理盐水,模型组腹腔静脉注射无血清的DMEM培养液,细胞移植组腹腔静脉注射脂肪间充质干细胞悬液。移植后2周,监测各组大鼠体质量、血糖水平,ELISA法检测各组大鼠胰岛素分泌量,RT-PCR检测胰腺组织PDX-1 m RNA的表达。结果与结论:(1)移植前细胞移植组与模型组的体质量均低于正常组,移植后2周细胞移植组大鼠体质量逐渐增加,而模型组的体质量持续下降;(2)与正常组比较,模型组大鼠空腹血糖维持在较高水平(P<0.05),与模型组相比,细胞移植组大鼠空腹血糖水平显著下降(P<0.05);(3)与正常组比较,模型组胰岛素水平明显降低(P<0.05),与模型组比较,细胞移植组胰岛素水平则显著增加(P<0.05);(4)正常组胰腺组织PDX-1 mR NA表达最高,模型组最低,各组间比较差异有显著性意义(P<0.05);(5)结果表明,脂肪间充质干细胞移植促进胰岛组织PDX-1表达,改善了大鼠的高血糖状态。