Apigenin ameliorates imiquimod-induced psoriasis in C57BL/6J mice by inactivating STAT3 and NF-κB

Psoriasis is a chronic autoimmune disease featured by patches on the skin.It is caused by malfunction of immune cells and keratinocytes with inflammation as one of its key features.Apigenin(API)is a natural flavonoid with anti-inflammatory and immunoregulatory properties.Therefore,we speculated that API can ameliorate psoriasis,and determined its effect on the development of psoriasis by using imiquimod(IMQ)-induced psoriasis mouse model.Our results showed that API attenuated IMQ-induced phenotypic changes,such as erythema,scaling and epidermal thickening,and improved splenic hyperplasia.Abnormal differentiation of immune cells was restored in API-treated mice.Mechanistically,we revealed that API is a key regulator of signal transducer activator of transcription 3(STAT3).API regulated immune responses by reducing interleukin-23(IL-23)/STAT3/IL-17A axis.Moreover,it suppressed IMQ-caused cell hyperproliferation by inactivating STAT3 through regulation of extracellular signal-regulated kinase 1/2 and nuclear factor-κB(NF-κB)pathway.Furthermore,API reduced expression of inflammatory cytokines through inactivation of NF-κB.Taken together,our study demonstrates that API can ameliorate psoriasis and may be considered as a strategy for psoriasis treatment.

Isoflavone Attenuates the Nuclear Transcription Factor Kappa B (NF-<i>κ</i>B) Activation on MPP⁺-Induced Apoptosis of PC12 Cells

Objective: To explore the underlying molecular mechanisms of cellular response to the challenge by 1-methyl-4-phenylpyridinium (MPP+)-induced apoptosis of PC12 cells, an in vitro cell model for Parkinson’s disease, and the effect of NF-κB activation on the protection of Parkinson’s disease by Isoflavone (I). Methods: PC12 cells were used to establish the cell model of Parkinson’s disease, and are divided into five groups: control group;MPP+ group;I (Isoflavone) + MPP+ group;I group;SN-50 + MPP+ group. The content of NF-κB in PC12 cells was determined by immunocytochemistry;The viability of PC12 cells after treated with cell-permeable NF-κB inhibitor SN-50 and cell viability were measured by MTT assay;the expression levels of NF-κB p65 in cytoplasm and nuclear fractions were evaluated by western blot analysis;the mRNA expression of NF-κB p65 was analyzed by in situ hybridization (ISH). Results: Compared with the control group, the protein of NF-κB p65 both in cytoplasm and in nuclei was significantly higher than in I + MPP+ and MPP+ groups;similarly, the mRNA expression level of NF-κB p65 gene was also significantly higher;moreover, the protein expression of NF-κB p65 was much lower in I group (P + group, the protein of NF-κB p65 was significantly lower in I + MPP+ group, the mRNA expression level of NF-κB p65 gene was also significantly lower, and the protein expression level of NF-κB p65 was much lower in I + MPP+ group (P + group (P > 0.05). Conclusion: NF-κB activation is essential to MPP+-induced apoptosis in PC12 cells;but Isoflavone can inhibit the cell damage to some extent to execute its protective function, which may be involved in nigral neurodegeneration in patients with Parkinson’s disease.

NF-κB和HMGB1在大鼠肠缺血再灌注肺损伤中的表达及白藜芦醇苷的保护作用

目的:探讨肠缺血再灌注(Intestinal ischemia-reperfusion,IIR)肺损伤核转录因子-κB(Nuclearfactor-kappaB,NF-κB)的活性和高迁移率族蛋白1(High mobility group box1,HMGB1)基因表达及白藜芦醇苷(Polydatin,PD)的保护作用。方法:54只Sprague-Dawley(SD)大鼠随机分成对照组、肠缺血45min继以再灌注组(再灌注0.5h、2h、6h)及肠缺血45min继以再灌注同时给予PD治疗组(再灌注0.5h、2h、6h)。采用比色法检测肺组织髓过氧化物酶(myeloperoxidase,MPO)水平、免疫组化法检测细胞间黏附分子-1(Intercellular adhesion molecule-1,ICAM-1)和NF-κB的表达、RT-PCR检测肺组织高迁移率族蛋白1(HMGB1)mRNA。结果:IIR后肺组织明显水肿、出血和炎性细胞侵润。与对照组相比,肺组织W/D、MPO水平明显升高,ICAM-1及NF-κB的蛋白表达,HMGB1mRNA(2h、6h)的表达增强(P<0.05);白藜芦醇苷干预后肺损伤程度减轻、各项指标明显改善(P<0.05)。结论:PD可以减轻肠缺血再灌注肺损伤,作用机制可能与其抑制NF-κB活化,降低肺组织HMGB1mRNA的表达上调密切相关。

siRNA阻断NF-κB信号通路抑制胃癌SGC-7901细胞的增殖及侵袭

目的采用RNA干扰(RNA interference,RNAi)技术下调胃癌细胞株SGC-7901中NF-κB p65基因的表达,探讨其对肿瘤细胞增殖活性和侵袭能力的影响。方法利用阳离子脂质体LipofectamineTM2000将化学合成的人NF-κB p65的小干扰RNA(small interference RNA,siRNA)转染入胃癌细胞株SGC-7901中。采用RT-PCR法测定细胞内NF-κB p65mRNA的表达;酶联免疫吸附测定法(ELISA)检测NF-κB亚单位p65的DNA结合活性的改变;采用MTT法测定细胞增殖活性的变化;利用Transwell侵袭实验检测细胞体外侵袭能力的变化。结果与对照组比较,化学合成的人NF-κB p65siRNA能有效地抑制SGC-7901细胞中NF-κB p65mRNA的表达(P<0.05);RelAsiRNA组的p65亚单位与DNA结合活性明显低于对照组(P<0.05),并且RelA siRNA组中SGC-7901细胞的体外侵袭力减弱,增殖活性降低。结论特异的siRNA可以有效阻断NF-κB信号通路,影响人胃癌细胞的增殖活性和体外侵袭能力。

火针对带状疱疹后遗神经痛大鼠疼痛阈值及NF-κB表达的影响

目的观察火针对带状疱疹后遗神经痛大鼠机械性痛阈值、脊髓核转录因子-κB(nuclear factor-κB,NF-κB)表达的影响,探讨火针治疗带状疱疹后遗神经痛的可能机制。方法将30只雄性SD大鼠,随机分为3组,即空白对照组、模型组、火针组,每组大鼠均为10只。通过VZV接种建立带状疱疹后神经痛(postherpetic neuralgia,PHN)动物模型。于接种前1 d,接种后1 d、4 d、7 d、14d和21 d分别进行机械性痛阈(paw withdrawal threshold,PWT)测定。接种后第7天开始,火针组进行火针治疗,隔日治疗1次,7次1个疗程。第21天,取各组大鼠L4、5段脊髓,用免疫组化法测定脊髓NF-κBp65蛋白表达情况。结果与空白对照组比较,模型组从造模后7 d开始出现PWT值下降(P<0.01),造模后第14天、21天时仍处于较低水平(P<0.01)。与模型组比较,火针组第7天治疗后开始出现PWT值上升(P<0.01),第14天、21天时PWT值上升明显(P<0.01)。与空白对照组比较,模型组脊髓组织中NF-κBp65表达明显增多,差异有统计学意义(P<0.01),与模型组比较,火针组脊髓组织中NF-κBp65表达明显减低,差异有统计学意义(P<0.01)。结论火针可缓解PHN大鼠PWT值,抑制NF-κB的表达,可能是火针治疗PHN的机制之一。

Effects of IκBα and its mutants on NF-κB and p53 signaling pathways

AIM： To study the effects of IκBα and its mutants （IκBαM, IκBα3N, IκBαM44C） on NF-κB, p53 and their downstream target genes. The relationship of NF-κB, p53, and IκBα was further discussed. METHODS： pECFP-IκBα, pECFP-IκBαM （amino acides 1-317, Ser32, 36A）, pECFP-IκBα243N （amino acides 1-243）, pECFP-IκBα244C （amino acides 24±317）, pEYFP-p65 and pp53-DsRed were constructed and transfected to ASTC-α-1 cells. Cells were transfected with pECFP-Cl as a control. 30 h after the transfection, location patterns of NF-κB, p53 and IκBα（IκBαM, IκBα243N, IκBα224C） were observed by a laser scanning microscope （LSM510/ConfoCor2, Zeiss）. RNA extraction and reverse transcription were performed in cells transfected or co-transfected with different plasmids. Effects of IκBα and its mutants on the transprition level of NF-κB, NF-κB downstream target gene TNF-α, p53 and p53 downstream target gene Bax were observed by real time QT-PCR. In all experiments β-actin was reference. Results are expressed as the target/reference ratio of the sample divided by the target/reference ratio of the control. Different transfected cells were incubated with CCK-8 for 2 h in the incubator. Then the absorbance at 450 nm was measured by using a microplate reader. RESULTS： Cells that were transfected with p53- DsRed revealed a predominant nuclear localization. YFP-p65 mainly existed in the cytoplasm. Cells were transfected with CFP-IκBα, CFP-IκBαM, and CFP-IκBα243N respectively and revealed a predominant cytosolic localization. However, cells transfected of CFP-IκBα244C revealed a predominant nuclear localization. The rnRNA levels of p65, TNF-α, p53 and Bax in CFP-IκBα transfected cells did not change significantly, while in YFP-p65/CFP-IκBα co-transfected cells, IκBα decreased the transcription of p65 downstream gene TNF-α （2.24 ± 0.503） compared with the YFP-p65/ CFP-C1 co-transfected cells （5.08 ± 0.891） （P 〈 0.05）. Phosphorylation defective IκBα （IκBαM） decreased the transcription levels of all the four genes compared with the control （P 〈 0.05）. The N terminus of IκBα（IκBα243N） increased the transcription of NF-κB （1.84 ± 0.176） and TNF-α （1.51 ± 0.203） a little bit. However, the C terminus of IκBα（IκBα244C） increased the transcription of NF-κB, TNF-α, p53 and Bax significantly （8.29 ± 1.662, 14.16 ± 2.121, 10.2 ± 0.621, 3.72 ± 0.346） （P 〈 0.05）. The CCK-8 experiment also showed that IκBα244C and p53 synergistically mediate apoptosis. CONCLUSIONS： IκBα and its mutants （IκBαM, IκBα243N, IκBαM244C） have different effects on NF- KB and p53 signaling pathways, according to their different structures. IκBαbounds with NF-KB and p53 in cytoplasm steadily, and inhibits both of the two signaling pathways, p53 and IκBα244C may be co-factor in inducing apoptosis. The C terminal of IκBαnhanced cell death, which suggests that it may be a pro-apoptotic protein existed in cells.

A Nonradioactive Method for Detecting DNA-binding Activity of Nuclear Transcription Factors

To determine the feasibility of a nonradioactive electrophoresis mobility shift assay for detecting nuclear transcription factor, double-stranded oligonucleotides encoding the consensus target sequence of NF-κB were labled with DIG by terminal transferase After nuclear protein stimulated with phorbol 12-myristate 13-acetate (PMA) or PMA and pyrrolidine dithiocarbamate (PDTC) electrophoresed on 8 % nondenaturing poliacrylamide gel together with oligeonucleotide probe, they were electro-blotted nylon membrane positively charged Anti-DIG-AP antibody catalyzed chemiluminescent substrate CSPD to image on X-film The results showed that nuclear proteins binded specifically to the NF-κB consensus sequence in the EMSA by chemiluminescent technique method and the activity of NF-κB in PMA group was more than that in PMA+PDTC group It is suggested that detection of NF-κB by EMSA with chemiluminescent technique is feasible and simple, which can be performed in ordinary laboratories

Promising Effects of Zerumbone on the Regulation of Tumor-promoting Cytokines Induced by TNF-α-activated Fibroblasts

Inflammation plays an important role in the development of several cancers.Inflammatory cytokines,including tumor necrosis factor-α(TNF-α),are associated with the induction of inflammation.Chronic inflammation contributes to the progression of cancer through several mechanisms,including increased cytokine production and activation of transcription factors,such as nuclear factor-κB(NF-κB).Zerumbone(ZER),a component of subtropical ginger(Zingiber zerumbet Smith),seems to have anti-inflammatory,anti-cancer,and antioxidant activities.In this study,we aimed to explore the protective function and mechanisms of ZER against TNF-α-induced cancer-promoting cytokines.We found that the viability of stimulated human fibroblast cell lines was reduced after treatment with ZER(IC50=18µmol/L),compared to un-stimulated fibroblasts(IC50=40µmol/L).Besides,ZER inhibited mRNA expression and protein secretion of transforming growth factor-β(TGF-β),interleukin-33(IL-33),monocyte chemoattractant protein-1(MCP-1),and stromal cell-derived factor 1(SDF-1),which were produced by TNF-α-induced fibroblasts,as measured by quantitative real time-PCR(qRT-PCR)and ELISA assays.The mRNA expression levels of TGF-β,IL-33,SDF-1,and MCP-1 showed 8,5,2.5,and 4-fold reductions,respectively.Moreover,secretion of TGF-β,IL-33,SDF-1,and MCP-1 was reduced to 3.65±0.34 ng/mL,6.3±0.26,1703.6±295.2,and 5.02±0.18 pg/mL,respectively,compared to the untreated group.In addition,the conditioned media(CM)of TNF-α-stimulated fibroblasts increased the NF-κB expression in colorectal cancer cell lines(HCT-116 and Sw48),while in the vicinity of ZER,the expression of NF-κB was reversed.Considering the significant effects of ZER,this component can be used as an appropriate alternative herbal treatment for cancer-related chronic inflammation.

EYA4通过抑制NF-κB依赖的RAP1反式激活抑制肝细胞癌的生长和侵袭

背景与目的我们之前研究表明,眼缺失蛋白同源物4(eyes absent homolog 4,EYA4)是果蝇眼部发育相关的眼缺乏蛋白家族成员之一,在肝细胞癌(hepatocellular carcinoma,HCC)标本中常发生甲基化和沉默,并与患者生存期短密切相关。本研究旨在探讨EYA4在HCC中作为肿瘤抑制因子的作用机制。方法转染EYA4表达质粒(pEYA4)构建稳定表达EYA4的人HCC细胞系Huh-7和PLC/PRF/5(PLC)。通过BALB/c裸鼠皮下注射稳定转染细胞建立异种移植肿瘤。组织标本来自75例病理诊断为HCC的患者。采用实时定量聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)、蛋白质免疫印迹和免疫组织化学的方法检测EYA4在细胞系、异种移植物和临床标本中的表达;研究了稳定转染细胞系的细胞增殖、克隆形成、侵袭性和肿瘤形成。利用基因表达芯片筛选EYA4调节的基因。通过共转染EYA4和带Flag标签的RAS相关蛋白1A(RAS-related protein 1A,RAP1A)基因的表达质粒(pEYA4和Flag-RAP1A)、功能研究、染色质免疫共沉淀、免疫荧光染色和细胞泛素化分析等方法,研究了EYA4对核因子-κB(nuclear factor-κB,NF-κB)/RAS相关蛋白1(RAS-related protein 1,RAP1)信号通路的影响。结果恢复HCC细胞系中EYA4的表达可抑制细胞的增殖、抑制克隆形成、降低细胞的侵袭性和抑制异种移植肿瘤生长,在体外实验中Flag-RAP1A可逆转pEYA4的抑制作用。用肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)激活NF-κB通路可增加p65与RAP1A基因启动子的结合并上调RAP1蛋白的表达。用BAY 11-7085和p65 siRNA抑制NF-κB通路,成功阻断了TNF-α诱导的RAP1上调。EYA4拮抗了TNF-α诱导的NF-κB抑制因子α(inhibitor of NF-κBα,IκBα)的磷酸化和泛素化以及p65的核易位和反式激活,进而抑制了NF-κB的活性和RAP1的表达。用calyculin A阻断EYA4的丝氨酸/苏氨酸磷酸酶活性可显著消除其对NF-κB活性的抑制作用。此外,EYA4的表达与HCC临床标本中IκBα/RAP1活性呈负相关。结论我们的研究结果为明确EYA4是真正的肿瘤抑制因子提供了功能和机制的理论基础,该肿瘤抑制因子可抑制NF-κB/RAP1信号通路的异常激活,从而抑制HCC生长和侵袭。

Nuclear Factor kappa B p65 Expression in Mouse Cochlea

Nuclear factor kappa B(NF-κB) is one of the best-characterized transcription factors playing important roles in many cellular responses to a large variety of stimuli,including inflammatory cytokines, phorbol esters, growth factors, and bacterial and viral products. The aim of this study is to demonstrate NF-κB expression in the mouse cochlea and its enhancement in response to lipopolysaccharides(LPS) and kanamycin(KA) treatment. Methods KA treatment consisted of subcutaneous KA injections at 700 mg/kg twice a day with an eight-hour interval between the two injections for 3 or 7 days. For animals in the LPS treatment group, a single dose of 0.3 mg LPS dissolved in 0.2 ml sterile saline were injected into both bullae through the tympanic membrane and kept there for 3 hours. Animals in the control group received subcutaneous saline injection for 7 days. Following immmunohistochemichal processing with rabbit polyclonal anti-NF-κB p65 antibodies, cryosections of the cochlea were examined for expression of NF-κB p65 in various structures in the cochlea. Results NF-κB p65 expression, identified by presence of brown reaction products characteristic of DAB immunohistochemistry, was visible in the spiral ligament, spiral prominence, tectorial membrane(TM), spiral ganglion and nerve fibers. Relatively weak NF-κB p65 expression was also visualized in the organ of Corti. Within the organ of Corti, the inner hair cells(IHC), outer hair cells(OHC), inner pillar cells(IP), outer pillar cells (OP), Deiter’s cells(DC), and Boettcher’s cells exhibited stronger staining than the inner sulcus cells, Hensen’s cells(HC) and Claudius’cells. No NF-κB p65 expression was seen in the nucleus of the IHC and OHC. NF-κB p65 expression was increased in animals exposed to LPS or KA, demonstrating significant differences in the staining between control animals and LPS/KA-treated animals. NF-κB p65 expression was not significantly different between LPS treated and KA treated animals or between 3 and 7 days in KA-treated animals. Conclusion LPS and KA exposure increases expression of NF-κB p65 in the mouse cochlea.

Lipopolysaccharide enhances the inhibition of NF-κB expression in NNK-mediated peritoneal macrophages

Objective： The aim of the study was to investigate the effect of lipopolysaccharide （LPS） on the expression of nuclear factor kappa B （NF-κB） in 4-（methylitrosamino）-1-（3-pyridyl）-1-butanone （NNK）-mediated primary mouse peritoneal macrophages in vitro. Methods： The activity of peritoneal rnacrophages treated with different concentrations of LPS was detected by MTT assay in rider to find the optimal concentration. Peritoneal macrophages were also treated with NNK （100-500 μM）, with or without LPS for 9 h. The expression of NF-κB was demonstrated via immunocytochemistry （ICC） and Western- blot, respectively. Results： The concentration of LPS at 25 μg/mL was found to be the optimal concentration to improve the activity of peritoneal macrophages （P 〈 0.01）. Simultaneously, LPS （25 μg/mL） increased the expression of NF-κB in both the nucleus and cytoplasm and facilitated transfer of NF-κB to the nucleus. NNK treatment significantly inhibited the expression of NF-κB in a concentration-dependent manner, among the LPS-stimulated or unstimulated peritoneal macrophages, especially when cotreated with LPS （25 μg/mL, P 〈 0.01 ）. Furthermore, NNK treatment （500 μM） with LPS yielded a significant decrease in NF-κB translocation to nucleus and inhibited the expression of NF-κB （P 〈 0.005）. Conclusion： LPS enhances the suppression of NF-κB expression in NNK-mediated mouse peritoneal macrophages, which may provide a theoretical basis for the inhibition of cancer.

胃癌中细胞核因子-κB、环氧合酶-2与血管内皮生长因子的表达及与血管生成的关系

目的探讨细胞核因子-κB(NF-κB)、环氧合酶-2(COX-2)与血管内皮生长因子(VEGF)在胃癌组织中的表达,分析三者的相关性及其与胃癌血管生成的关系。方法收集南京市江宁医院2010年至2011年中晚期胃癌手术切除病理组织67例标本,其中男性43例,女性24例;年龄42～65岁,平均年龄50岁。采用免疫组织化学SP法检测67例胃癌标本中NF-κB、COX-2及VEGF的表达情况,以相应癌旁组织(55例,男性32例,女性23例;年龄43～68岁,平均年龄52.5岁)作为对照,并且采用抗CD34抗体标记微血管内皮细胞,计算微血管密度(MVD)。结果 NF-κB、COX-2及VEGF在胃癌中的阳性表达率分别为62.69%、64.18%及79.10%,且三者的表达均呈正相关,胃癌组织中NF-κB、COX-2及VEGF表达均阳性组,MVD值也最高(35.95±3.38),与三者表达均阴性组相比,MVD值的差异具有统计学意义(P<0.05)。结论在胃癌组织中,NF-κB、COX-2及VEGF均高表达,三者均可能参与了胃癌的血管生成,并对胃癌的血管生成起促进作用。

阻断 NF -κB 信号途径防治急性肺损伤的研究进展

目的：探讨以核因子κB（NF-κB）为靶点防治急性肺损伤（ALI）的分子生物学机制。方法应用CNKI、Medline、ScienceDirect 等数据库，查阅近年来相关文献，总结NF-κB在ALI中的作用机制与干预手段。结果 NF-κB在ALI中的信号转导途径及作用机制逐渐被揭示，大量研究证实，通过干预NF-κB上游信号通路、NF-κB抑制蛋白（ IκB）、IκB激酶（ IKK）等途径，能在一定限度内阻断细胞因子和炎症介质的释放，缓解ALI的炎症反应。结论阻断靶细胞中NF-κB通路、特异性抑制目的蛋白和基因表达，或许能成为未来治疗ALI的研究方向；不恰当的药物干预会破坏机体的免疫平衡状态，虽然NF-κB阻断剂已进入临床试验阶段，但多数仅局限于动物实验和细胞水平，实现临床防治尚需进行大量实验研究。

dsODNs 靶向封闭肺泡巨噬细胞中 NF -κB炎症信号通路的实验研究

目的：在确定脂质体转染双链寡聚脱氧核苷酸（ dsODNs ）最佳转染浓度及时间的基础上，评估脂质体转染的双链寡聚脱氧核苷酸（ dsODNs/Lipofectamin2000）竞争性抑制对肺泡巨噬细胞中核因子κB（ NF-κB）/DNA结合活性的影响，验证dsODNs-decoy策略靶向封闭肺泡巨噬细胞中NF-κB信号通路的可行性。方法支气管肺泡灌洗分离提取家兔肺泡巨噬细胞（ AMs）后体外培养，以Lipofectamine2000为载体转染dsODNs后对巨噬细胞进行干预，测定巨噬细胞中dsODNs/Lipofectamine2000的转染活性、转染效率及转染后对AMs的毒性；检测dsODNs/Lipofectamine2000转染对炎症因子（ IL -1α、IL -6、TNF -α等） mRNA 的表达；观察 dsODNs/Lipofectamine2000转染后NF -κB p65亚基的核移位情况。结果转染肺泡巨噬细胞dsODNs/Lipofectamine2000的最适比例为1∶5，最佳时间为6 h，此时细胞毒性适中，转染效率、荧光强度及细胞状态最佳；与LPS组比较，dsODNs/Lipofectamine2000转染组各种炎症介质mRNA的表达均明显抑制（P＜0．01）；细胞核中NF-κB p65亚基的表达明显少于细胞浆中。结论 dsODNs/Lipofecetamine2000能在体外有效转染AMs并成功抑制AMs中NF-κB信号通路相关炎症靶基因的转录表达，证实了dsODNs-decoy策略影响炎症效应细胞的可行性。

Hydrogen sulfide protects against amyloid beta-peptide induced neuronal injury via attenuating inflammatory responses in a rat model

Neuroinflammation has been recognized to play a critical role in the pathogenesis of Alzheimer＇s disease （AD）, which is pathologically characterized by the accumulation of senile plaques containing activated microglia and amyloid β-peptides （Aβ）. In the present study, we examined the neuroprotective effects of hydrogen sulfide （H2S） on neuroinflammation in rats with Aβ1-40 hippocampal injection. We found that Aβ-induced rats exhibited a disorder of pyramidal cell layer arrangement, and a decrease of mean pyramidal cell number in the CA1 hippocampal region compared with those in sham operated rats. NaHS （a donor of H2S, 5.6 mg/kg/d, i.p.） treatment for 3 weeks rescued neuronal cell death significantly. Moreover, we found that H2S dramatically suppressed the release of TNF-α, IL-1β and IL-6 in the hippocampus. Consistently, both immunohistochemistry and Western blotting assays showed that H2S inhibited the upregulation of COX-2 and the activation of NF-κB in the hippocampus. In conclusion, our data indicate that H2S suppresses neuroinflammation via inhibition of the NF-κB activation pathway in the Aβ-induced rat model and has potential value for AD therapy.

Albumin resuscitation protects against traumatic/hemorrhagic shock-induced lung apoptosis in rats

Objective: To determine the effects of albumin administration on lung injury and apoptosis in traumatic/hemorrhagic shock (T/HS) rats. Methods: Studies were performed on an in vivo model of spontaneously breathing rats with induced T/HS; the rats were subjected to femur fracture, ischemia for 30 min, and reperfusion for 20 min with Ringer's lactate solution (RS) or 5% (w/v) albumin (ALB), and the left lower lobes of the lungs were resected. Results: Albumin administered during reperfusion markedly attenuated injury of the lung and decreased the concentration of lactic acid and the number of in situ TdT-mediated dUTP nick-end labelling (TUNEL)-positive cells. Moreover, immunohistochemistry performed 24 h after reperfusion revealed increases in the level of nuclear factor κB (NF-κB), and phosphorylated p38 mitogen-activated protein kinase (MAPK) in the albumin-untreated group was down-regulated by albumin treatment when compared with the sham rats. Conclusion: Resuscitation with albumin attenuates tissue injury and inhibits T/HS-induced apoptosis in the lung via the p38 MAPK signal transduction pathway that functions to stimulate the activation of NF-κB.

Influence of catgut implantation at acupoints on splenic lymphocyte nuclear factor (NF-κB p65) and correlated signaling molecules (β2AR) in rats with experimental colitis

Objective To investigate the mechanisms of catgut implantation at acupoints on ulcerative colitis. Methods Eighteen SD rats were randomly divided into a normal control group （NC）, a model group （MO） and a catgut implantation group （CI） with 6 rats in each group. Animals in group MO and group CI were treated by trinitro-benzene-sulfonic acid （TNBS） to establish model with colitis. No other treatment was given to the rats in group MO, but catgut was implanted at ＂Shàngjùxū＂ （上 巨虚 ST 37）, ＂Tiānshū＂ （天枢 ST 25） and ＂Dàchángshū＂ （大肠俞 BL 25） in the rats in group CI. The symptoms of diarrhea and bloody stool, and changes in histopathology were detected 15 days after the treatment. Expressions of splenic lymphocyte nuclear factor κB p65（NF-κB p65）and correlated signaling molecules（β2AR）were detected by the western blot method. Results Diarrhea and mucus bloody purulent stool were soon controlled, and mucous injures were obviously improved in group CI. The NF-κB p65 value of splenic lymphocytes was signifi cantly increased （P0.01） and expression of β2AR remarkably reduced in group MO （P0.01）, compared with group NC. But, the NF-κB p65 value was significantly decreased （P0.01） and expression of β2AR remarkably increased in group CI （P 0.01） , compared with group MO. Conclusion Catgut implantation at acupoints is obviously effective in treating experimental colitis. Modulation of NF-κB p65 and the correlated signaling molecules β2AR may be involved in the mechanisms.

Scutellarein Ameliorated Chondrocyte Inflammation and Osteoarthritis in Rats

Objective:Osteoarthritis(OA)is a degenerative joint disorder characterized by the gradual degradation of joint cartilage and local inflammation.This study aimed to investigate the anti-OA effect of scutellarein(SCU),a single-unit flavonoid compound obtained from Scutellaria barbata D.Don,in rats.Methods:The extracted rat chondrocytes were treated with SCU and IL-1β.The chondrocytes were divided into control group,IL-1βgroup,IL-1β+SCU 50µmol/L group,and IL-1β+SCU 100µmol/L group.Morphology of rat chondrocytes was observed by toluidine blue and safranin O staining.CCK-8 method was used to detect the cytotoxicity of SCU.ELISA,qRT-PCR,Western blotting,immunofluorescence,SAβ-gal staining,flow cytometry,and bioinformatics analysis were applied to evaluate the effect of SCU on rat chondrocytes under IL-1βintervention.Additionally,anterior cruciate ligament transection(ACL-T)was used to establish a rat OA model.Histological changes were detected by safranin O/fast green,hematoxylin-eosin(HE)staining,and immunohistochemistry.Results:SCU protected cartilage and exhibited anti-inflammatory effects via multiple mechanisms.Specifically,it could enhance the synthesis of extracellular matrix in cartilage cells and inhibit its degradation.In addition,SCU partially inhibited the nuclear factor kappa-B/mitogen-activated protein kinase(NF-κB/MAPK)pathway,thereby reducing inflammatory cytokine production in the joint cartilage.Furthermore,SCU significantly reduced IL-1β-induced apoptosis and senescence in rat chondrocytes,further highlighting its potential role in OA treatment.In vivo experiments revealed that SCU(at a dose of 50 mg/kg)administered for 2 months could significantly delay the progression of cartilage damage,which was reflected in a lower Osteoarthritis Research Society International(OARSI)score,and reduced expression of matrix metalloproteinase 13(MMP13)in cartilage.Conclusion:SCU is effective in the therapeutic management of OA and could serve as a potential candidate for future clinical drug therapy for OA.

蠲痹历节清方对改良痛风性关节模型大鼠滑膜的TLR4,NF-κB,PPARγ的影响

目的：观察蠲痹历节清方对改良痛风性关节炎大鼠滑膜组织中Toll样受体4（Toll-like receptors 4,TLR4）,核转录因子kappa B（nuclear factor-kappa B,NF-κB）,过氧化物酶体增殖物激活受体γ（peroxisome proliferator-activated receptorγ,PPARγ）的影响,探讨蠲痹历节清方治疗急性痛风性关节炎局部组织炎症的可能作用机制。方法：将42只SD雄性大鼠,按随机数字表法选取6只为正常组,30只为造模组,造模组采用改良痛风性关节炎造模后随机分为模型组,蠲痹历节清方高、中、低剂量组（4 400,2 200,1 100 mg·kg^-1）,依托考昔组（11 mg·kg^-1）,吡格列酮组（20 mg·kg^-1）,每组6只。正常组及模型组的大鼠以生理盐水灌胃20 mL·kg^-1。每组每只大鼠每日灌胃给药2次,连续2 d后处死大鼠,取受试右踝关节,分离出滑膜组织,分为两部分,一部分用于病理形态学观察,一部分用逆转录聚合酶链式反应（RT-PCR）检测TLR4,NF-κB p65,PPARγmRNA的表达水平,蛋白质免疫印迹法（Western blot）检测TLR4,NF-κB p65,PPARγ蛋白表达水平。结果：与正常组比较,模型组中TLR4和NF-κB mRNA和蛋白表达显著升高（P〈0.01）,PPARγmRNA和蛋白表达升高（P〈0.05）;与模型组比较,吡格列酮组及蠲痹历节清方高、中、低剂量组TLR4和NF-κB mRNA和蛋白的表达显著降低（P〈0.01）,PPARγmRNA和蛋白的表达显著升高（P〈0.05）。结论：蠲痹历节清方可能通过上调PPARγ表达,抑制TLR4,NF-κB表达从而抑制急性痛风性关节炎局部组织炎症。

有氧运动与饮食干预对肥胖小鼠睾丸氧化应激的影响

目的:通过对肥胖小鼠施加有氧运动与饮食干预,探索运动与饮食干预对肥胖小鼠睾丸氧化应激和p38MAPK-NF-κB通路中的作用。方法:随机将17只C57BL/6J小鼠分为正常饮食组(ND),37只分为高脂饮食组(HFD),高脂饮食脂肪占比40%,喂养12周后,HFD组剔除3只肥胖抵抗小鼠,其余34只肥胖造模成功;随后将ND组分为正常饮食对照组(NC,n=8),正常饮食运动组(NE,n=9),肥胖高脂饮食对照组(OC,n=8),肥胖高脂饮食运动组(OE,n=9),肥胖正常饮食组(ONC,n=8),肥胖正常饮食运动组(ONE,n=9),各组继续饲养8周,其中NE、OE和ONE组以速度20 m/min,60 min/d,6 d/week,进行8周跑台运动,末次运动后36~40 h取血和睾丸组织,ELISA检测血清睾酮和睾丸氧化应激(MDA、T-SOD、T-AOC)水平,RT-PCR和Western blot检测睾丸p38MAPK-NF-κB水平。结果:与NC组比较,OC组小鼠体脂参数、睾丸MDA和睾丸p38MAPK-NF-κB mRNA和蛋白水平明显升高(P<0.01),睾丸SOD、睾丸系数和血睾酮明显降低(P<0.01);NE组小鼠体脂参数明显降低(P<0.05),血清睾酮明显升高(P<0.01)。与OC组比较,OE组小鼠体脂参数、睾丸MDA和睾丸p38MAPK-NF-κB mRNA和蛋白水平明显降低(P<0.05或0.01),睾丸SOD和血睾酮水平明显升高(P<0.01);ONC组小鼠体脂参数、睾丸MDA和睾丸p38MAPK-NF-κB mRNA和蛋白水平明显降低(P<0.01),睾丸SOD水平和睾丸系数明显升高(P<0.05);ONE组小鼠体脂参数、睾丸MDA和睾丸p38MAPK-NF-κB mRNA和蛋白水平明显降低(P<0.01),睾丸SOD、睾丸系数和血睾酮水平明显升高(P<0.01)。结论:肥胖引起小鼠睾丸发生氧化应激,上调睾丸p38MAPK-NF-κB水平,并降低血睾酮水平;运动、饮食和运动×饮食干预均能通过降低体脂,改善睾丸氧化应激,下调睾丸p38MAPK-NF-κB水平。