The Role of Toll-Like Receptors and Nuclear Factor κB p65 Protein in the Pathogenesis of Otitis Media

The role of Toll-like receptor 4 (TLR4) and nuclear factor κB p65 (NF-κB p65) proteins in the pathogenesis of otitis media is explored. In recent years, the incidence of otitis media has been rising globally, becoming a significant threat to human health. More and more studies have found that Toll-like receptor 4 (TLR4), as a member of the Toll-like receptor family, can promote the generation of inflammatory factors and is closely related to the body’s immune response and inflammatory response. Nuclear factor-κB p65 (NF-κB p65) is a nuclear transcription factor that can interact with various cytokines, growth factors, and apoptotic factors, participating in processes such as oxidative stress, apoptosis, and inflammation in the body [1]. This article elaborates on the structure, function, and signaling pathways of TLR4 and NF-κB p65 proteins in the pathogenesis of otitis media, aiming to provide more precise targets and better therapeutic efficacy for the diagnosis and treatment of otitis media. The role of inflammation in disease.

褪黑素调控GPX4/NF-κB p65信号通路干预铁死亡促进小鼠皮肤创面修复

目的:探讨褪黑素对人皮肤成纤维细胞(HSF)增殖和铁死亡的作用,并探究其促进小鼠皮肤创面修复的分子机制。方法:采用CCK-8法检测HSF细胞增殖能力,选取细胞增殖率最高的褪黑素浓度进行后续实验。将HSF细胞随机分为对照组、谷氨酸组、谷氨酸+褪黑素组。谷氨酸+褪黑素组用100μmol/L褪黑素预处理细胞60 min,随后除对照组外,另两组加入谷氨酸(10 mmol/L)诱导铁死亡。采用相应试剂盒检测HSF细胞活性氧(ROS)、谷胱甘肽(GSH)、丙二醛(MDA)、Fe^(2+)水平。构建小鼠为全层皮肤损伤模型,并随机分为对照组和褪黑素组,每组10只。褪黑素组在皮肤损伤处外敷含5 mg褪黑素的凡士林乳膏,对照组外敷等量凡士林乳膏,每日清洁换药,连续9 d。计算术后第5、7、9天各组创面愈合率。采用HE和Masson染色观察第5、9天小鼠创面组织病理学变化及胶原纤维生成情况。采用免疫荧光染色法检测术后第9天创面组织中细胞增殖情况,Western blot检测术后第9天创面组织中谷胱甘肽过氧化物酶4(GPX4)、核因子(NF)-κB p65、p-NF-κB p65蛋白表达。结果:褪黑素浓度为100μmol/L时HSF细胞增殖率最高,故选取100μmol/L褪黑素进行后续实验。与对照组比较,谷氨酸组HSF细胞GSH水平降低,ROS、MDA、Fe^(2+)水平升高(均P<0.05)。与谷氨酸组比较,谷氨酸+褪黑素组HSF细胞GSH水平升高,ROS、MDA、Fe^(2+)水平降低(均P<0.05)。与对照组比较,褪黑素组小鼠术后第5、7、9天创面愈合率增加(均P<0.05)。与对照组比较,褪黑素组小鼠术后第5、9天创面组织再上皮化程度更高,胶原纤维生成比例增加(均P<0.05)。与对照组比较,褪黑素组小鼠术后第9天创面组织Ki-67免疫荧光强度增加(P<0.05)。与对照组比较,褪黑素组小鼠术后第9天创面组织GPX4蛋白表达水平升高,NF-κB p65、p-NF-κB p65蛋白表达水平降低(均P<0.05)。结论:褪黑素能够促进皮肤成纤维细胞增殖,抑制细胞铁死亡,促使小鼠创面组织再上皮化和胶原纤维形成,加快创面愈合,其机制可能与调控GPX4/NF-κB p65信号通路有关。

慢性肾脏病NF-κB p65和炎性因子与动脉硬化的关系及HDF治疗对其影响

目的 探讨慢性肾脏病(CKD)患者体内核因子(NF)-κB p65和相关炎性因子水平与并发动脉硬化之间的关系及进行血液滤过透析(HDF)治疗对其产生影响。方法 随机选取肾功能正常的CKD1期患者(N组)40例N组,血液净化中心维持性血液透析(HD)CKD5D期患者80例,分别测定血清NF-κB p65和人单核细胞趋化蛋白(MCP)-1水平、超敏C反应蛋白(hs-CRP)、白细胞介素(IL)-6、IL-8及肿瘤坏死因子(TNF)-α水平,同时B超测定患者颈动脉内-中膜厚度(IMT),以判断是否并发动脉硬化;之后CKD5D期患者随机分为在线(OL)HDF组及HD组,分别以OL HDF及HD进行治疗,治疗3个月后两组复测以上指标进行比较并再次进行颈动脉超声检查测定IMT。结果 与N组相比,HD组、HDF组治疗前血清IL-6、IL-8、TNF-α、hs-CRP、MCP-1、NF-κB p65水平均明显增高(P<0.01);NF-κB p65蛋白表达明显增高,发生动脉硬化比例也明显增高(P<0.05)。经3个月治疗后,HD组血清IL-6、IL-8、TNF-α、hs-CRP、MCP-1、NF-κB水平,NF-κB p65蛋白表达均无明显变化(P>0.05),仍明显高于N组,但HDF组上述指标较治疗前相比明显改善(P<0.05),NF-κB p65蛋白表达受抑制。3个月后HD组动脉硬化发生率明显增高(P<0.05),而HDF组减少,但无统计学差异(P>0.05)。结论 HDF治疗下调NF-κB p65蛋白表达,能明显减轻慢性肾衰患者的微炎症状态,减少动脉硬化的发生率,是维持性HD患者的必要治疗。

Nuclear factor-kB p65 (RelA) transcription factor is constitutively activated in human colorectal carcinoma tissue

AIM: Activation of transcription factor nuclear factor-κB(NF-κB) has been shown to play a role in cell proliferation, apoptosis, cytokine production, and oncogenesis. The purpose of this study was to determine whether NF-κB was constitutively activated in human colorectal tumor tissues and, if so, to determine the role of NF-κB in colorectal tumorigenesis, and furthermore, to determine the association of RelA expression with tumor cell apoptosis and the expression of Bcl-2 and Bcl-xL. METHODS: Paraffin sections of normal epithelial, adenomatous and adenocarcinoma tissues were analysed immunohistochemically for expression of RelA, Bcl-2 and Bcl-xL proteins. Electrophoretic mobility shift assay (EMSA) was used to confirm the increased nuclear translocation of RelA in colorectal tumor tissues. The mRNA expressions of Bcl-2 and Bcl-xL were determined by reverse transcription polymerase chain reaction (RT-PCR) analysis. Apoptotic cells were detected triphosphate fluorescence nick end labeling (TUNEL) method. RESULTS: The activity of NF-κB was significantly higher in adenocardnoma tissue in comparison with that in adenomatous and normal epithelial tissues. The apoptotic index (AI) significantly decreased in the transition from adenoma to adenocarcinoma. Meanwhile, the expressions of Bcl-2 and Bcl-xL protein and their mRNAs were significantly higher in adenocarcinoma tissues than that in adenomatous and normal epithelial tissues. CONCLUSION: NF-κB may inhibit apoptosis via enhancing the expression of the apoptosis genes Bcl-2 and Bcl-xL. And the increased expression of RelA/nuclear factor-κB plays an important role in the pathogenesis of colorectal carcinoma.

Mesalazine alleviated the symptoms of spontaneous colitis in interleukin-10 knockout mice by regulating the STAT3/NF-κB signaling pathway

BACKGROUND Excessive endoplasmic reticulum(ER)stress in intestinal epithelial cells can lead to damage to the intestinal mucosal barrier,activate the signal transducer and activator of transcription 3(STAT3)/nuclear factor kappa B(NF-κB)signaling pathway,and exacerbate the inflammatory response,thus participating in the pathogenesis of ulcerative colitis(UC).Mesalazine is a commonly used drug in the clinical treatment of UC.However,further studies are needed to determine whether mesalazine regulates the ER stress of intestinal epithelial cells,downregulates the STAT3/NF-κB pathway to play a role in the treatment of UC.AIM To study the therapeutic effects of mesalazine on spontaneous colitis in interleukin-10(IL-10)-/-mice.METHODS The 24-week-old IL-10-/-mice with spontaneous colitis were divided into the model group and the 5-amino salicylic acid group.Littermates of wild-type mice of the same age group served as the control.There were eight mice in each group,four males and four females.The severity of symptoms of spontaneous colitis in IL-10-/-mice was assessed using disease activity index scores.On day 15,the mice were sacrificed.The colon length was measured,and the histopathological changes and ultrastructure of colonic epithelial cells were detected.The protein expressions of STAT3,p-STAT3,NF-κB,IκB,p-IκB,and glucoseregulated protein 78 were identified using Western blotting.The STAT3 and NF-κB mRNA expressions were identified using real-time polymerase chain reaction.The glucose-regulated protein 78 and C/EBP homologous protein expressions in colon sections were detected using immunofluorescence.RESULTS Mesalazine reduced the symptoms of spontaneous colitis in IL-10 knockout mice and the histopathological damage of colonic tissues,and alleviated the ER stress in epithelial cells of colitis mice.Western blotting and quantitative real-time polymerase chain reaction results showed that the STAT3/NF-κB pathway in the colon tissue of model mice was activated,suggesting that this pathway was involved in the pathogenesis of UC and might become a potential therapeutic target.Mesalazine could down-regulate the protein expressions of p-STAT3,NF-κB and p-IκB,and down-regulate the mRNA expression of STAT3 and NF-κB.CONCLUSION Mesalazine may play a protective role in UC by reducing ER stress by regulating the STAT3/NF-κB signaling pathway.

TPPU通过抑制p38 MAPK/NF-κB p65信号通路对阿尔茨海默病细胞模型的抗神经炎症作用

目的在Aβ25-35诱导的阿尔茨海默病(AD)小胶质细胞(BV2细胞)模型中,采用对BV2细胞进行干预,探讨1-三氟甲氧基苯基-3-(1-丙酰哌啶-4-基)脲(TPPU),TPPU是否具有抗神经炎症作用及其可能的抗炎机制。方法利用Aβ25-35作用BV2细胞构建AD细胞模型,CCK8法分别检测不同浓度Aβ25-35和TPPU处理对BV2细胞活性的影响,最终选择20μM Aβ25-35诱导BV2细胞48 h作为造模组,0.1μM TPPU预处理BV2细胞3h,20μM Aβ25-35诱导BV2细胞48 h作为治疗组进行后续实验。利用ELISA法检测丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性,倒置荧光显微镜检测活性氧(ROS)产生,real-time PCR检测小胶质细胞M1表型促炎因子肿瘤坏死因子(TNF)、白细胞介素-1β(IL-1β)基因mRNA表达水平和检测M2表型抗炎因子IL-10及标志物Arg1基因mRNA表达水平。通过Western blot法检测细胞中炎症因子TNF-α蛋白及p38 MAPK/NF-κB p65信号通路相关蛋白表达水平。结果与对照组相比,模型组BV2细胞活力降低(P<0.05),ROS显著升高(P<0.05),MDA含量显著增多(P<0.05),SOD活性显著下降(P<0.05),TNF mRNA、IL-1βmRNA表达水平明显上调(P<0.05),IL-10 mRNA、Arg1 mRNA表达水平明显下调(P<0.05),TNF-α蛋白及p-p38 MAPK、p-NF-κB p65蛋白表达明显上调(P<0.05)。与模型组相比,经TPPU预处理后,BV2细胞活力明显升高(P<0.05),ROS显著降低(P<0.05),MDA含量显著下降(P<0.05),SOD活性显著提高(P<0.05),TNF mRNA、IL-1βmRNA表达水平明显下调(P<0.05),IL-10 mRNA、Arg1 mRNA表达水平明显上调(P<0.05),TNF-α蛋白及p-p38 MAPK、p-NF-κB p65蛋白表达明显下调(P<0.05)。结论TPPU抑制Aβ25-35诱导的BV2细胞炎症反应,促进BV2细胞由M1表型向M2表型极化,其机制可能与抑制p38MAPK/NF-κB p65信号通路激活有关。

腹主动脉瘤患者血清转胶蛋白、基质金属蛋白酶9和核因子-κB p65水平检测及意义

目的:检测腹主动脉瘤(AAA)患者血清转胶蛋白(TAGLN)、基质金属蛋白酶9(MMP-9)和核因子-κB(NF-κB)p65水平变化,探讨其临床意义。方法:通过酶联免疫吸附法检测肾下AAA患者80例(AAA组)及体检健康者50例(健康组)血清TAGLN、MMP-9和NF-κB p65水平并进行比较。分析三者间的相关性,分析三者对AAA的诊断价值。结果:健康组血清TAGLN水平高于AAA组,MMP-9和NF-κB p65水平低于AAA组(均P<0.05)。血清TAGLN与MMP-9和NF-κB p65呈负相关(r=-0.196、-0.364,均P<0.05),MMP-9与NF-κB p65呈正相关(r=0.213,P=0.015)。TAGLN、MMP-9和NF-κB p65联合诊断AAA的曲线下面积(AUC)高于任意单独指标,而敏感度和特异度不低于任意单独指标(均P<0.05)。结论:AAA患者血清TAGLN水平升高,MMP-9和NF-κB p65水平降低,三者联合检测对AAA的诊断具有较高价值。

超早期小骨窗微创术治疗高血压脑出血疗效及对核因子-κB p65通路的影响

目的:分析超早期小骨窗微创术治疗高血压脑出血的疗效及对核因子-κB p65(NF-κB p65)通路的影响。方法:选取高血压脑出血86例,根据手术时间不同分为观察组(46例,发病6 h内进行小骨窗微创颅内血肿清除术)和对照组(40例,发病6~24 h内进行小骨窗微创颅内血肿清除术)。比较两组临床疗效、围手术期指标、脑血流灌注指标、脑组织创伤应激指标、NF-κB p65通路相关因子以及并发症发生情况。结果:与对照组比较,观察组总有效率更高(P<0.05)。与对照组比较,观察组术中出血量更高,但意识恢复时间、下床时间、住院时间更短(均P<0.05)。与术前比较,术后7 d两组大脑中动脉舒张压末期血流速度(EDV)、收缩期峰值流速(PSV)水平以及血清脑源性神经营养因子(BDNF)水平升高,且观察组更高;两组大脑中动脉阻力指数(RI)、患侧颈总动脉外周阻力(R)降低,血清血清神经肽Y(NPY)、星形胶质源性蛋白(S100β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、NF-κB p65水平降低,且观察组更低(均P<0.05)。观察组并发症总发生率低于对照组(P<0.05)。结论:相对于发病6~24 h进行手术,发病6 h内的超早期小骨窗微创术疗效更好,不但可以改善患者围手术期指标,降低创伤应激,改善脑血流灌注,还可以调控NF-κB p65通路,安全性更高。

Targeting GPR65 alleviates hepatic inflammation and fibrosis by suppressing the JNK and NF-κB pathways

Background:G-protein coupled receptors(GPCRs)are recognized as attractive targets for drug therapy.However,it remains poorly understood how GPCRs,except for a few chemokine receptors,regulate the progression of liver fibrosis.Here,we aimed to reveal the role of GPR65,a proton-sensing receptor,in liver fibrosis and to elucidate the underlying mechanism.Methods:The expression level of GPR65 was evaluated in both human and mouse fibrotic livers.Furthermore,Gpr65-deficient mice were treated with either bile duct ligation(BDL)for 21 d or carbon tetrachloride(CCl4)for 8 weeks to investigate the role of GPR65 in liver fibrosis.A combination of experimental approaches,including Western blotting,quantitative real-time reverse transcription-polymerase chain reaction(qRT-PCR),and enzyme-linked immunosorbent assay(ELISA),confocal microscopy and rescue studies,were used to explore the underlying mechanisms of GPR65’s action in liver fibrosis.Additionally,the therapeutic potential of GPR65 inhibitor in the development of liver fibrosis was investigated.Results:We found that hepatic macrophage(HM)-enriched GPR65 was upregulated in both human and mouse fibrotic livers.Moreover,knockout of Gpr65 significantly alleviated BDL-and CCl4-induced liver inflammation,injury and fibrosis in vivo,and mouse bone marrow transplantation(BMT)experiments further demonstrated that the protective effect of Gpr65knockout is primarily mediated by bone marrow-derived macrophages(BMMs).Additionally,in vitro data demonstrated that Gpr65 silencing and GPR65 antagonist inhibited,while GPR65 overexpression and application of GPR65 endogenous and exogenous agonists enhanced the expression and release of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and transforming growth factor-β(TGF-β),all of which subsequently promoted the activation of hepatic stellate cells(HSCs)and the damage of hepatocytes(HCs).Mechanistically,GPR65 overexpression,the acidic pH and GPR65 exogenous agonist induced up-regulation of TNF-αand IL-6 via the Gαq-Ca^(2+)-JNK/NF-κB pathways,while promoted the expression of TGF-βthrough the Gαq-Ca^(2+)-MLK3-MKK7-JNK pathway.Notably,pharmacological GPR65 inhibition retarded the development of inflammation,HCs injury and fibrosis invivo.Conclusions:GPR65 is a major regulator that modulates the progression of liver fibrosis.Thus,targeting GPR65 could be an effective therapeutic strategy for the prevention of liver fibrosis.

Inhibition of p38 mitogen-activated protein kinase attenuates experimental autoimmune hepatitis: Involvement of nuclear factor kappa B

To investigate the role of p38 mitogen-activated protein kinase （p38MAPK） in murine experimental autoimmune hepatitis （EAH）.METHODS： To induce EAH, the syngeneic S-100 antigen emulsified in complete Freud＇s adjuvant was injected intraperitoneally into adult male C57BI/6 mice. Liver injury was assessed by serum ALT and liver histology. The expression and activity of p38 MAPK were measured by Western blot and kinase activity assays. In addition, DNA binding activities of nuclear factor kappa B （NF-KB） were analyzed by electrophoretic mobility shift assay. The effects of SB203580, a specific p38 MAPK inhibitor, on liver injuries and expression of proinflammatory cytokines （interferon-y, IL-12, IL-1β and TNF-α） were observed.RESULTS： The activity of p38 MAPK and NF-~：B was increased and reached its peak 14 or 21 d after the first syngeneic S-100 administration. Inhibition of p38 MAPK activation by SB203580 decreased the activation of NF-~：B and the expression of proinflammatory cytokines. Moreover, hepatic injuries were improved significantly after SB203580 administration.

Effects of ω-3 fatty acids on toll-like receptor 4 and nuclear factor-κB p56 in lungs of rats with severe acute pancreatitis

AIM To determine the effects of ω-3 fatty acids(ω-3FA) on the toll-like receptor 4(TLR4)/nuclear factor κB p56(NF-κBp56) signal pathway in the lungs of rats with severe acute pancreatitis(SAP).METHODS A total of 56 Sprague-Dawley rats were randomly divided into 4 groups: control group, SAP-saline group, SAP-soybean oil group and SAP-ω-3FA group. SAP was induced by the retrograde infusion of sodium taurocholate into the pancreatic duct. The expression of TLR4 and NF-κBp56 in the lungs was evaluated by immunohistochemistry and Western blot analysis. The levels of inflammatory cytokines interleukin-6 and tumor necrosis factor-alpha in the lungs were measured by enzyme-linked immunosorbent assay. RESULTS The expression of TLR4 and NF-κBp56 in lungs and of inflammatory cytokines in serum significantly increased in the SAP group compared with the control group(P < 0.05), but was significantly decreased in the ω-3FA group compared with the soybean oil group at 12 and 24 h(P < 0.05).CONCLUSION During the initial stage of SAP, ω-3FA can efficiently lower the inflammatory response and reduce lung injury by triggering the TLR4/NF-κBp56 signal pathway.

Clinical significance of SQSTM1/P62 and nuclear factor-κB expression in pancreatic carcinoma

BACKGROUND Overexpression of SQSTM1(sequestosome 1,P62)and nuclear factor-κB(NF-κB)plays an important role in the invasion and metastasis of a variety of malignant tumors.AIM To explore the expression of P62 and NF-κB in pancreatic cancer and their relationship with clinicopathological features.METHODS The expression levels of P62 and NF-κB were analyzed by immunohistochemistry with a tissue chip containing 40 cases of human pancreatic carcinoma.Then we analyzed the correlation among P62 expression,phospho-P65 expression,and clinicopathological features of pancreatic carcinoma samples.RESULTS P62 expression was mainly observed in the cytoplasm of pancreatic carcinoma cells.Phosphorylated P65(phospho-P65)was mainly expressed in the nucleus and cytoplasm of pancreatic carcinoma cells.There was a significant difference in P62 expression among T stages.And a significant difference in phosphor-P65 expression among pathology types was noted.In the cases with strongly positive P62 expression,significant differences were found in age.And there were significant differences in T stage and tumor-node-metastasis stage in the cases with strongly positive phosphor-P65 expression.CONCLUSION In pancreatic carcinoma,P62 expression is significantly correlated with T stage.It may be a valuable malignant indicator for human pancreatic carcinoma.

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.

Effect of NF-κB p65 antisense oligodeoxynucleotide on transdifferentiation of normal human lens epithelial cells induced by transforming growth factor-β2

AIM：To study the inhibition of nuclear factor kappa-B p65（NF-κB p65）antisense oligodeoxynucleotide（ASODN）on transdifferentiation of normal human lens epithelial cells induced by transforming growth factor-β2（TGF-β2）.·M ETHODS：NF-κBp65ASODNand NF-κBp65missense oligodeoxynucleotide（MSODN）were designed and synthesized.Human lens epithelial cell line（HLE B-3）cells were prepared for study and divided into 7 groups.Control group was HLE B-3 cells cultured in dulbecco’s modified eagle medium（DMEM）.T1,T2,and T3 group were HLE B-3 cells cultured in DMEM with 10 ng/m L TGF-β2 for 6h,12h,24h respectively.A＋T group was HLE B-3 cells cultured with 10 ng/m L TGF-β2for 24h after transfected by NF-κB p65 ASODN for 24h.M＋T group was HLE B-3 cells cultured with 10 ng/m L TGF-β2 for 24h after transfected by NF-κB p65 MSODN for 24h.The negative control group was HLE B-3 cells cultured with 10 ng/m L TGF-β2 for 24h after cultured with transfer agent（Hi Per Fect）for 24h.Cell morphology was observed at different time points using an inverted microscope.The expression of NF-κB p65 m RNA was detected with reverse transcription-polymerase chain reaction（RT-PCR）,and the expression ofα-smooth muscle actin（α-SMA）protein was assayed with ELISA.·RESULTS：With the TGF-β2 stimulation prolongation,the expression of NF-κB p65 m RNA and a-SMA protein increased in T1,T2,T3 groups compared with the control group,and the difference was statistically significant（〈0.05）.NF-κB p65 ASODN lowered the expression of NF-κB p65 m RNA andα-SMA protein induced by TGF-β2.NF-κB p65 MSODN and Hi Per Fect did not lower the expression of NF-κB p65 m RNA andα-SMA protein induced by TGF-β2.The difference between control group and A＋T group was not statistically significant（〉0.05）,but the difference among A＋T group and other groups was statistically significant（〈0.05）.·CONCLUSION：NF-κB p65 ASODN could lower the expression of NF-κB p65 m RNA andα-SMA protein induced by TGF-β2,and antagonized TGF-β2-induced transdifferentiation of HLE B-3.NF-κB p65ASODN could be used as a new biological therapeutic target of posterior capsular opacification.

参芪补肺汤对COPD肺气虚证大鼠气道平滑肌HDAC2与NF-κB p65表达的影响

目的观察参芪补肺汤对慢性阻塞性肺疾病(COPD)肺气虚证大鼠支气管平滑肌(ASM)增殖中组蛋白去乙酰化酶-2(HDAC2)和核因子κB(NF-κB)p65表达的影响。方法将40只SD雄性大鼠随机分为正常对照组、模型对照组、参芪补肺汤组、氨茶碱组,每组各10只。采用气管内滴注脂多糖加烟熏28 d的方法建立COPD肺气虚证模型。光镜下观察肺组织病理形态学变化,图像分析法测量小气道管壁和ASM厚度,采用免疫组化、实时荧光定量PCR和Wester blot方法检测大鼠ASM中HDAC2和NF-κB p65表达。结果与正常对照组比较,模型对照组大鼠气道管壁和ASM明显增厚(P<0.05),NF-κB p65 mRNA和蛋白表达明显增高(P<0.05),HDAC2 mRNA和蛋白表达明显降低(P<0.05)。与模型对照组比较,参芪补肺汤组和氨茶碱组气道管壁和ASM厚度明显降低(P<0.05),NF-κB p65 mRNA和蛋白表达明显降低(P<0.05),HDAC2mRNA和蛋白表达均明显升高(P<0.05),参芪补肺汤组与氨茶碱组比较,均差异无统计学意义(P>0.05)。结论参芪补肺汤可抑制COPD肺气虚证模型大鼠ASM增殖,其机制与其提高HDAC2表达、减少NF-κB p65表达有关。

参芪补肺汤对慢性阻塞性肺疾病肺气虚证大鼠气道平滑肌中乙酰化组蛋白H4、HDAC2和NF-μB p65表达的影响

目的观察参芪补肺汤对慢性阻塞性肺疾病(COPD)肺气虚证大鼠支气管平滑肌(ASM)中乙酰化组蛋白H4、组蛋白去乙酰化酶-2(HDAC2)和核因子-κB p65(NF-κB p65)表达的影响。方法取SD大鼠40只,随机分为正常组、模型组、参芪补肺汤组、氨茶碱组,每组10只。运用气管内注射脂多糖加烟熏28 d的方法建立COPD肺气虚证大鼠模型。光镜下观察肺组织的病理形态学变化,运用图像分析法测量小气道管壁和ASM的厚度,采用免疫组化和Western blot方法检测大鼠ASM中乙酰化组蛋白H4、HDAC2和NF-κB p65的蛋白表达,实时荧光定量PCR方法检测大鼠ASM组织中HDAC2 m RNA和NF-κB p65 m RNA的表达。结果与正常组比较,模型组大鼠气道管壁和ASM厚度明显增高(P<0.05);与模型组比较,参芪补肺汤组和氨茶碱组气道管壁和ASM厚度明显降低(P<0.05);参芪补肺汤组与氨茶碱组比较差异无统计学意义(P>0.05)。与正常组比较,模型组乙酰化组蛋白H4的蛋白表达、NF-κB p65 m RNA和蛋白的表达明显增高(P<0.05);HDAC2m RNA和蛋白的表达均明显降低(P<0.05);与模型组比较,参芪补肺汤组和氨茶碱组乙酰化组蛋白H4的蛋白表达、NF-κB p65 m RNA和蛋白的表达明显增高(P<0.05);HDAC2 m RNA和蛋白的表达明显降低(P<0.05)。参芪补肺汤组与氨茶碱组比较差异均无统计学意义(P>0.05)。结论参芪补肺汤可抑制COPD肺气虚证模型大鼠ASM增殖,其机制与其提高HDAC2的表达,使组蛋白H4去乙酰化,从而抑制NF-κB p65的表达有关。

脉络膜新生血管模型大鼠视网膜核转录因子-κB p65、血管内皮生长因子和碱性成纤维细胞生长因子表达的变化

目的观察脉络膜新生血管(choroidal neovascularization,CNV)模型大鼠视网膜核转录因子-κB p65(nuclear transcription factor kappa-B p65,NF-κB p65)、血管内皮生长因子(vascular endothelial growth factor,VEGF)、碱性成纤维细胞生长因子(basic fibroblast growth factor,b FGF)表达的变化。方法取BN大鼠12只,随机分为正常组、模型组,每组各6只。正常组不做任何处理,模型组以100 g·L^(-1)水合氯醛腹腔注射麻醉,复方托吡卡胺滴眼液双眼散瞳,倍诺喜行眼表麻醉,在全视网膜镜下用532 nm激光于视盘周围做视网膜光凝建立CNV模型。每周行FFA、OCT检查眼底,观察CNV生成情况。5周后处死大鼠,取大鼠视网膜用免疫组织化学法检测视网膜内NF-κB p65、VEGF、b FGF阳性表达积分光密度。结果造模5周后FFA及OCT检查可见模型组CNV形成。免疫组织化学法检测结果:正常组大鼠视网膜上NF-κB p65阳性表达的积分光密度为9264.33±1479.49,模型组的积分光密度为34 815.83±3873.61,两组比较差异有统计学意义(t=15.095,P<0.01);正常组大鼠视网膜上VEGF阳性表达的积分光密度为1994.93±309.00,模型组为13 318.54±1958.11,两组比较差异有统计学意义(t=13.992,P<0.01);正常组大鼠视网膜上b FGF阳性表达的积分光密度为1608.74±235.55,模型组为15 963.14±2034.12,两组比较差异有统计学意义(t=17.171,P<0.01)。结论 532 nm激光光凝可诱导大鼠视网膜上NF-κB p65的表达,NF-κB p65在CNV发生发展中可能起重要作用。

苍附导痰汤对肥胖型PCOS-IR模型大鼠卵巢TLR4/NF-κB p65信号通路的影响

目的 探讨苍附导痰汤对肥胖型PCOS-IR (polycystic ovarian syndrome-insulin resistance, PCOS-IR)模型大鼠卵巢Toll受体4(TLR4)/核转录因子κB p65(NF-κB p65)信号通路的调控作用。方法 48只♀大鼠随机分为正常组8只和模型组40只。来曲唑(1 mg·kg^(-1))联合高脂饮食建立肥胖型PCOS-IR大鼠模型,快速革兰染色法观察动情周期,挑选24只模型大鼠随机分为:模型组、阳性药(二甲双胍135 mg·kg^(-1))组、苍附导痰汤高、低剂量(57.96、14.49 g·kg^(-1))组,每组各6只,药物干预21 d。观察动情周期、卵巢和子宫指数变化;血液生化仪测定空腹血糖(FBG)和血脂(甘油三酯TG和总胆固醇TC)变化;酶联免疫吸附(ELISA)法测定空腹胰岛素(FINS)水平;免疫组化法和荧光定量PCR法检测卵巢中TLR4和NF-κB p65蛋白及基因的表达。结果 与正常组比较,模型组大鼠动情周期紊乱,卵巢多囊性改变明显,FBG、TG、TC含量和FINS、HOMA-IR水平上调,卵巢中TLR4和NF-κB p65蛋白及mRNA表达均增加(P<0.05);与模型组比较,苍附导痰汤高剂量组大鼠排卵周期得到改善,卵巢多囊性改变减轻,上述指标出现明显逆转(P<0.05)。结论 苍附导痰汤能有效改善肥胖型PCOS-IR大鼠卵巢排卵和糖脂代谢功能,作用机制可能与调控卵巢TLR4/NF-κB p65信号通路有关。

游离脂肪酸刺激核因子NF-kBp65核转位诱导3T3-L1脂肪细胞胰岛素抵抗的分子机制

目的：研究游离脂肪酸对3T3-L1脂肪细胞核因子NF—κBp65表达及转位的影响，探讨游离脂肪酸诱导胰岛素抵抗的分子机制．方法：诱导成熟的3T3-L1脂肪细胞与0.3，0．5，1．0mmol／L的软脂酸（PA）培养6—24h，用葡萄糖氧化酶法检测培液中的葡萄糖消耗量，以2-脱氧-[^3H]-D-葡萄糖摄入法观察葡萄糖的转运率，用Western blot检测总NF—κBp65蛋白及核NF—κBp65蛋白的表达，用激光扫描共聚焦（CLSM）对NF—κBp65进行定位显示．结果：0．3—1．0mmol／L软脂酸作用6—24h后，3T3-L1脂肪细胞的葡萄糖消耗明显减少（3．03±0．34，2．71±0．36，2．64±0．25mmol／L），呈时间剂量依赖效应，其作用不需要胰岛素的存在：0．3—1．0mmol／L软脂酸作用6—24h显著减少3T3-L1脂肪细胞胰岛素刺激的葡萄糖转运率（64％，33％，32％），呈时间剂量依赖效应；核NF-κBp65蛋白表达明显增加，CLSM显示NF-κBp65核转位增加．但软脂酸对3T3-L1脂肪细胞总NF—κBp65蛋白的表达无明显影响．结论：游离脂肪酸可以诱导胰岛素抵抗，其分子机制可能与FFAs刺激NF—κB的活化转位调节相关基因的表达有关．

NF-κB p65对子宫内膜异位症腺上皮细胞OPN、MMP-9表达及细胞侵袭性的影响

目的:探讨RNAi抑制核因子NF-κB p65对子宫内膜异位症(EMs)在位内膜腺上皮细胞中OPN、MMP-9表达及细胞侵袭性的影响。方法:采用NF-κB p65 siRNA基因沉默转染10例EMs患者在位内膜原代腺上皮细胞,由Western blot、Real-time PCR、Transwell等分别检测干扰前后OPN、NF-κB p65、MMP-9蛋白和mRNA表达及细胞侵袭性的变化。结果:与未干预组比较,NF-κB p65 siRNA干预后EMs腺上皮细胞中OPN蛋白及mRNA表达无明显变化,差异无统计学意义(P均>0.05);NF-κB p65、MMP-9蛋白及mRNA表达降低,差异均有统计学意义(P均<0.05)。NF-κB p65 siRNA干预后,EMs腺上皮细胞的细胞侵袭性明显降低,差异有统计学意义(P<0.05)。结论:揭示NF-κB可能通过下调MMP-9的表达进而改变细胞侵袭性诱导EMs的发生。