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.

TLR4/NF-κB p65高表达对皮肤鳞状细胞癌增殖、侵袭和转移的影响

目的探讨与分析Toll样受体4(Toll-like recepTor 4,TLR4)/核因子KappaB(NF-κB)p65高表达对皮肤鳞状细胞癌增殖、侵袭和转移的影响。方法对数生长期的人皮肤鳞状细胞癌A431细胞株分为三组,空白组、对照组与实验组,分别转染DMEM培养基、pc3.0空载体、pc3.0-TLR4NF-κB p65载体,检测皮肤鳞状细胞癌增殖、侵袭和转移表达情况。结果转染24 h、48 h后,实验组的TLR4、NF-κB p65 RNA表达水平显著高于空白组与对照组,差异有统计学意义(P<0.05),空白组与对照组比较,差异无统计学意义(P>0.05)。实验组的细胞增殖指数显著低于空白组与对照组,差异有统计学意义(P<0.05),对照组与空白组比较,差异无统计学意义(P>0.05)。实验组的细胞侵袭指数与细胞转移指数都明显低于空白组与对照组,差异有统计学意义(P<0.05),对照组与空白组比较,差异无统计学意义(P>0.05)。实验组的Caspase-9/Myc蛋白表达水平显著高于空白组与对照组,差异有统计学意义(P<0.05),空白组与对照组比较,差异无统计学意义(P>0.05)。结论TLR4/NF-κB p65高表达能抑制皮肤鳞状细胞癌细胞的增殖、转移与侵袭,也可促进Caspase-9/Myc蛋白的表达,从而发挥促进肿瘤细胞凋亡的作用。

Homer1a reduces inflammatory response after retinal ischemia/reperfusion injury

Elevated intraocular pressure(IOP)is one of the causes of retinal ischemia/reperfusion injury,which results in NRP3 inflammasome activation and leads to visual damage.Homerla is repo rted to play a protective role in neuroinflammation in the cerebrum.However,the effects of Homerla on NLRP3inflammasomes in retinal ischemia/reperfusion injury caused by elevated IOP remain unknown.In our study,animal models we re constructed using C57BL/6J and Homer1^(flox/-)/Homerla^(+/-)/Nestin-Cre^(+/-)mice with elevated IOP-induced retinal ischemia/repe rfusion injury.For in vitro expe riments,the oxygen-glucose deprivation/repe rfusion injury model was constructed with M uller cells.We found that Homerla ove rexpression amelio rated the decreases in retinal thickness and Muller cell viability after ischemia/reperfusion injury.Furthermore,Homerla knockdown promoted NF-κB P65^(Ser536)activation via caspase-8,NF-κB P65 nuclear translocation,NLRP3 inflammasome formation,and the production and processing of interleukin-1βand inte rleukin-18.The opposite results we re observed with Homerla ove rexpression.Finally,the combined administration of Homerla protein and JSH-23 significantly inhibited the reduction in retinal thickness in Homer1^(flox/-)Homer1a^(+/-)/Nestin-Cre^(+/-)mice and apoptosis in M uller cells after ischemia/reperfusion injury.Taken together,these studies demonstrate that Homer1a exerts protective effects on retinal tissue and M uller cells via the caspase-8/NF-KB P65/NLRP3 pathway after I/R injury.

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.

中药对湿热型溃疡性结肠炎小鼠NFκB p65及免疫功能的影响

目的:探讨清热利湿方对湿热型溃疡性结肠炎(ulcerative colitis,UC)小鼠不同时段结肠黏膜核转录因子κB(NFκB)p65蛋白表达及相关免疫功能的影响。方法:选用80只SPF级雄性Balb/c小鼠,随机分为对照组、造模组、西药组、中药组,每组20只,除对照组外,其余3组采用2,4,6-三硝基苯磺酸(TNBS)/乙醇溶液灌肠法配合饮食加环境复合法建立小鼠湿热型UC模型,术后西药组予以美沙拉嗪灌肠,中药组予以清热利湿方灌肠;第3 d、7 d每组各取10只小鼠观察血清肿瘤坏死因子-α(TNF-α)、IgA、黏膜sIgA水平,结肠黏膜NFκB p65蛋白表达情况。结果:造模组TNF-α、IgA、黏膜sIgA水平升高,NFκB p65蛋白表达增加,与对照组有显著差异(P<0.01);与造模组相比,中药组、西药组TNF-α、IgA、黏膜sIgA、NFκB p65蛋白表达下降(P<0.01);中药组与西药组相比,给药3 d,上述各指标无明显差异(P>0.05)。给药7 d,中药组TNF-α为(240.16±16.24)pg/mL、IgA为(161.46±28.75)ng/mL、黏膜sIgA为(18.25±2.20)μg/mL、NFκB p65蛋白表达阳性率为33.3%,较西药组TNF-α(290.58±18.33)pg/mL、IgA(184.90±33.23)ng/mL、黏膜sIgA(20.86±1.84)μg/mL、NFκB p65蛋白表达阳性率(55.6%)明显降低(P<0.05)。结论:湿热型UC动物模型造模成功,清热利湿方治疗湿热型UC疗效肯定,作用机制可能与抑制NFκB p65蛋白表达、降低TNF-α、IgA、黏膜sIgA水平等相关免疫指标有关,随疗程增加,中药疗效优于西药。

膀胱移行细胞癌组织中NF-κB p65、uPA、Bcl-2的表达变化及其意义

目的观察膀胱移行细胞癌(下称膀胱癌)组织中NF-κB p65和尿激酶型纤溶酶原激活物(uPA)及Bcl-2的表达变化,并探讨其相关性。方法采用免疫组化SP法对40例膀胱癌组织和10例正常膀胱组织中NF-κBp65、uPA、Bcl-2进行测定,分析三者与膀胱癌分级、分期、侵袭转移及复发的关系及NF-κB p65表达与uPA、Bcl-2的相关性。结果 NF-κB p65、uPA、Bcl-2在膀胱癌组织中的表达均明显高于正常膀胱组织(P<0.05);NF-κB p65和uPA与膀胱癌病理分级、分期、淋巴结转移有关(P均<0.05),Bcl-2与膀胱癌病理分级、分期有关(P<0.05),与淋巴结转移无关(P>0.05);膀胱癌组织中,NF-κB p65表达与uPA、Bcl-2表达呈正相关(r分别为0.388、0.462,P均<0.05)。结论在膀胱癌组织中NF-κB p65、uPA、Bcl-2均呈高表达,并与膀胱癌的临床病理学特征有关;NF-κB p65可能通过调控uPA、Bcl-2的表达促进膀胱癌的进展;NF-κB p65、uPA、Bcl-2可以作为判断膀胱癌侵袭性、转移及预后的生物学标志物。

OPN、IKK-β、NF-κB p65和MMP-9在胃癌细胞中的表达及意义

目的:检测骨桥蛋白(osteopontin,OPN)、IκB激酶抑制剂-β(inhibitor of IκB kinase-beta,IKK-β)、核因子-κB p65(nuclear factor-κB p65,NF-κB p65)和基质金属蛋白酶-9(matrix metalloproteinases-9,MMP-9)在具有转移潜能和低分化的胃癌细胞系及永生化胃上皮细胞中的表达,为进一步研究OPN在胃癌侵袭转移中的可能作用机制及其临床应用提供实验基础。方法:免疫细胞化学SP法和Western blot检测OPN、IKK-β、NF-κB p65和MMP-9在具有转移潜能和低分化的胃癌细胞系MKN45、GC9811-C11、803、AGS、GC7901及永生化胃上皮细胞293中的表达。结果:(1)免疫细胞化学结果显示:胃癌细胞胞浆及胞周可见OPN和MMP-9弱阳性染色,尤以803明显。而在293细胞中,未见OPN和MMP-9阳性染色。IKK-β和NF-κB p65在胃癌细胞胞浆中均成阳性染色,胞核中还可见NF-κB p65弱阳性染色,而293细胞中IKK-β和NF-κB p65未见阳性染色。(2)Western blot结果显示:OPN、IKK-β和MMP-9在胃癌细胞MKN45、GC9811-C11、803、AGS、GC7901与293细胞中的表达有差异,细胞核中NF-κB p65的表达也有差异。结论:OPN、IKK-β、NF-κB p65和MMP-9与胃癌的侵袭转移密切相关,OPN参与胃癌侵袭转移的分子机制及其能否作为胃癌侵袭转移的生物学指标还需进一步的实验研究。

NF-κB/P65,p-IκBα,IκKα在皮肤SCC和BCC中的表达和意义

目的:探讨NF-κB信号转导通路中真核细胞转录因子κ(BNF-κB/P65)、磷酸化NF-κB抑制蛋白(p-IκBα)以及真核细胞转录因子抑制蛋白激酶(IκKα)在皮肤SCC和BCC中的表达和三者之间关系,为临床治疗提供新思路。方法:采用免疫组化SP法检测53例皮肤癌组织(其中SCC27例,BCC26例)和20例正常皮肤组织中NF-κB/P65,p-IκBα和IκKα蛋白的表达。结果:与正常皮肤组织相比,NF-κB/P65,p-IκBα和IκKα三者在SCC,BCC中均高表达(P<0.05);三者在SCC和BCC中的表达具有统计学意义(P<0.05);IκK蛋白的表达与SCC的分化程度无明显相关性(P>0.05)。在BCC和SCC中,IκKα的表达与p-IκBα的表达呈正相关关系(r=0.536,P<0.05),NF-κB/P65和p-IκBα的表达呈正相关关系(r=0.435,P<0.05)。结论:NF-κB信号传导通路可能在BCC和SCC的发生中起重要作用。

Role of nuclear factor kappa B in central nervous system regeneration

Activation of nuclear factor kappa B （NF-κB） is a hallmark of various central nervous system （CNS） pathologies. Neuron-specific inhibition of its transcriptional activator subunit RelA, also referred to as p65, promotes neuronal survival under a range of conditions, i.e., for ischemic or excitotoxic insults. In macro- and microglial cells, post-lesional activation of NF-κB triggers a growth-permissive program which contributes to neural tissue inflammation, scar formation, and the expression of axonal growth inhibitors. Intriguingly, inhibition of such inducible NF-~B in the neuro-glial compartment, i.e., by genetic ablation of RelA or overexpression of a trans- dominant negative mutant of its upstream regulator IκBa, significantly enhances functional recovery and promotes axonal regeneration in the mature CNS. By contrast, depletion of the NF-κB subunit p50, which lacks transcriptional activator function and acts as a transcriptional repressor on its own, causes precocious neuronal loss and exacerbates axonal degeneration in the lesioned brain. Collectively, the data imply that NF-κB orchestrates a multicellular pro- gram in which κB-dependent gene expression establishes a growth-repulsive terrain within the post-lesioned brain that limits structural regeneration of neuronal circuits. Considering these subunit-specific functions, interference with the NF-κB pathway might hold clinical potentials to improve functional restoration following traumatic CNS injury.

Oxymatrine Improves TNBS-induced Colitis in Rats by Inhibiting the Expression of NF-κB p65

Inflammatory bowel disease is thought to be regulated by the balance between Th1 and Th2 cytokines secreted by T cells, and NF-κB p65 also plays a predominant role in the intestinal inflammation. We evaluated the potency of oxymatrine, one of active components of Sophora Root, in inhibiting the immune responses and inflammation in 2,4,6-trinitrobenzene sulfonic acid （TNBS）-induced colitis. The inflammation was markedly ameliorated in the oxymatrine-treated rats. The level of IL-2 was increased and that of IL-10 was decreased in colon tissue in the rat model, which was reversed by the treatment of oxymatrine. Moreover, the elevated expression of NF-κB p65 in colon tissue in the model was also improved by oxymatrine treatment. Our results suggest that oxymatrine might be beneficial for the abnormal immune responses and inflammation by regulating the unbalance of Th1 and Th2 cytokines secretion and inhibiting the expression of NF-κB p65 in colon tissue.

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.

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.

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.

Suppressing high mobility group box-1 release alleviates morphine tolerance via the adenosine5'-monophosphate-activated protein kinase/heme oxygenase-1 pathway

Opioids,such as morphine,are the most potent drugs used to treat pain.Long-term use results in high tolerance to morphine.High mobility group box-1(HMGB1) has been shown to participate in neuropathic or inflammatory pain,but its role in morphine tolerance is unclear.In this study,we established rat and mouse models of morphine tolerance by intrathecal injection of morphine for 7 consecutive days.We found that morphine induced rat spinal cord neurons to release a large amount of HMGB1.HMGB1 regulated nuclear factor κB p65 phosphorylation and interleukin-1β production by increasing Toll-like receptor 4receptor expression in microglia,thereby inducing morphine tolerance.Glycyrrhizin,an HMGB1 inhibito r,markedly attenuated chronic morphine tole rance in the mouse model.Finally,compound C(adenosine 5’-monophosphate-activated protein kinase inhibitor) and zinc protoporphyrin(heme oxygenase-1 inhibitor)alleviated the morphine-induced release of HMGB1 and reduced nuclear factor κB p65 phosphorylation and interleukin-1β production in a mouse model of morphine tolerance and an SH-SY5Y cell model of morphine tole rance,and alleviated morphine tolerance in the mouse model.These findings suggest that morphine induces HMGB1 release via the adenosine 5’-monophosphate-activated protein kinase/heme oxygenase-1 signaling pathway,and that inhibiting this signaling pathway can effectively reduce morphine tole rance.

CBP基因沉默对大鼠颈动脉球囊损伤后内膜增生的影响

目的观察小干扰RNA(siRNA)重组慢病毒介导的CREB结合蛋白(CBP)基因沉默对大鼠颈动脉球囊损伤后新生内膜增生的影响,并探讨其作用机制。方法雄性SD大鼠48只,随机分为4组:假手术组、PBS对照组、慢病毒介导CBP基因siRNA转染组(CBP-siRNA-Lenti组)及慢病毒介导非CBP同源序列siRNA转染组(NC-siRNA-Lenti组),建立大鼠颈动脉球囊损伤模型,术后28天处死动物。分别用实时定量PCR、Western Blot检测大鼠颈动脉CBP和乙酰化核因子κB p65(NF-κB p65)的表达水平;病理组织学观察血管内膜增生情况;免疫组织化学染色对损伤血管壁增殖细胞核抗原(PCNA)的表达进行评估。结果术后28天,与PBS对照组和NC-siRNA-Lenti组比较,CBP-siRNA-Lenti组CBP mRNA和蛋白的表达显著下调(P均<0.05),CBP沉默能明显抑制新生内膜面积(0.108±0.008 mm2比0.238±0.022 mm2、0.252±0.016 mm2,P<0.05)、内膜与中膜面积比(0.706±0.062比1.483±0.136、1.497±0.137,P<0.05)的增加,及下调血管壁乙酰化NF-κB p65和PCNA的表达水平(P均<0.05)。结论慢病毒介导的CBP基因沉默能有效地抑制颈动脉球囊损伤后新生内膜的形成,其机制可能与抑制NF-κB p65的过度乙酰化有关。

Losartan reduced connexin43 expression in left ventricular myocardium of spontaneously hypertensive rats

Objective:To assess the effect of angiotensin II type 1(AT1)receptor antagonist losartan on myocardium con- nexin43(Cx43)gap junction(GJ)expression in spontaneously hypertensive rats(SHRs)and investigate possible mechanisms. Methods:Sixteen 9-week-old male SHRs and 8 age-matched male Wistar-Kyoto(WKY)rats were included in this study.SHRs were randomly divided into two groups to receive losartan at 30 mg/(kg·d)by oral gavage once daily for 8 weeks(SHR-L)or vehicle(0.9%saline)to act as controls(SHR-V);WKY rats receiving vehicle for 8 weeks served as normotensive controls.At the end of the experiment,rats were sacrificed and the hearts were removed.Expressions of Cx43 and nuclear factor-kappaB p65 (NF-κB p65)proteins in all three groups were observed and further investigations on the effect of angiotensin II type 1 receptor antagonist losartan(30 mg/(kg·d),8 weeks)on Cx43 expression were conducted with Western blot and immunohistochemistry. NF-κB p65 protein in nuclear extracts was determined by Western blot.Results:Left ventricular(LV)hypertrophy was prominent in SHRs,Cx43 and NF-κB p65 protein expressions were obviously upregulated and Cx43 distribution was dispersed over the cell surface.Treatment with losarton reduced the over-expressions of Cx43 and NF-κB p65 in LV myocardium.The distribution of Cx43 gap junction also became much regular and confined to intercalated disk after losartan treatment.Conclu- sion:Cx43 level was upregulated in LV myocardium of SHR during early stage of hypertrophy.Angiotensin II type 1 receptor antagonist losartan prevented Cx43 gap junction remodeling in hypertrophied left ventricles,possibly through the NF-κB pathway.

Curcumin suppresses gastric NF-κB activation and macromolecular leakage in Helicobacter pylori-infected rats

AIM:To investigate whether curcumin could attenuate nuclear factor(NF)-κB p65 expression and macromolecular leakage in the gastric mucosa of Helicobacter pylori(H.pylori)-infected rats.METHODS:Twenty-five male Sprague-Dawley rats were equally divided into five groups:control rats(Control),control rats supplemented with 600 mg/kg curcumin,H.pylori-infected rats(Hp),H.pylori-infected rats supplemented with 200 mg/kg curcumin(Hp + curIn H.pylori-infected groups,rats were inoculated with H.pylori suspension twice a day at an interval of 4 h for 3 d.Two weeks later,200 or 600 mg/kg curcumin was given once daily to curcuminsupplemented groups for 7 d.On the day of the experiment,macromolecular leakage in gastric mucosa was examined by intravital fluorescence microscopy.The stomach tissue was removed to examine NF-κB p65 expression in gastric epithelial cells by immunohistochemistry.RESULTS:The expression of NF-κB p65 in gastric epithelial cells and the macromolecular leakage from gastric mucosal microcirculation significantly increased in the Hp group compared with the Control group.The percentages of NF-κB p65 immunoreactive cells in Control and Hp groups were 10.72% ± 2.10% vs 16.02% ± 2.98%,P = 0.004,respectively.The percentages of macromolecular leakage in Control and Hp groups were 10.69% ± 1.43% vs 15.41% ± 2.83%,P = 0.001,respectively.Curcumin supplementation in Hp + cur-CONCLUSION:H.pylori-induced gastric inflammation in rats is associated with increased NF-κB activation and macromolecular leakage which can be reduced by curcumin supplementation.

Role of moxibustion in inflammatory responses during treatment of rat ulcerative colitis

AIM: To investigate the efficacy of moxibustion in ulcerative colitis (UC) rats from morphological, immunological and molecular biological perspectives. METHODS: Thirty-two Sprague-Dawley rats were randomly assigned to a blank control group (normal rats, n = 6) and a model replication (MR) group (UC rats, n = 26). A UC model was established by 2,4,6-trinitrobenzenesulfonic acid/dextran sulfate sodium enema. Rats in the MR group were further randomly assigned to a 9-min moxibustion (9M) group (9 moxa-cone, n = 6), 6-min moxibustion (6M) group (6 moxa-cone, n = 6), 3-min moxibustion (3M) group (3 moxa-cone, n = 6), and a waiting list control (WLC) group (no moxibustion treatment, n = 6). Rats in the moxibustion treatment group were treated in 14 sessions over 28 d. Disease activity, local tissue morphology, serum level of interleukin (IL)-8 and IL-10, and expression of Toll-like receptor (TLR)9 as well as nuclear factor (NF)-kappa B p65 in colonic tissue were determined by disease activity index (DAI), hematoxylin and eosin staining, electron microscopy, enzyme-linked immunosorbent assay and Western blotting, respectively. RESULTS: DAI was lowest in the 9M group and highest in the WLC group. The differences in DAI between the moxibustion treatment (3M, 6M, 9M) and no treatment groups were significant for all one-to-one comparisons (0.60 +/- 0.54 vs 1.20 +/- 0.44, 0.60 +/- 0.54 vs 1.80 +/- 0.45, 0.60 +/- 0.54 vs 3.0 +/- 0.45, respectively, P < 0.05). Light and electron microscopy showed that the neatness of the glandular arrangement in colonic mucosal epithelia gradually increased in the WLC, 3M, 6M to 9M groups. IL-8 level successively decreased while IL-10 level increased from the WLC to 3M, 6M and 9M groups. The differences among these groups were significant for all comparisons (105.46 +/- 8.75 vs 76.61 +/- 3.58, 105.46 +/- 8.75 vs 69.78 +/- 1.87, 105.46 +/- 8.75 vs 67.41 +/- 1.84, respectively, P < 0.01 for IL-8; and 30.83 +/- 1.29 vs 75.64 +/- 1.90, 30.83 +/- 1.29 vs 80.90 +/- 3.16, 30.83 +/- 1.29 vs 83.46 +/- 2.37, respectively, P < 0.01 for IL-10), except comparison of 6M vs 9M. Expression of TLR9 and NF-kappa B p65 decreased in order: highest in the WLC group and lowest in the 9M group. In addition, the differences among the WLC, 3M, 6M and 9M groups were significant for all comparisons (0.492 +/- 0.026 vs 0.380 +/- 0.022, 0.492 +/- 0.026 vs 0.355 +/- 0.005, 0.492 +/- 0.026 vs 0.327 +/- 0.015, respectively, P < 0.05 for TLR9; and 0.436 +/- 0.041 vs 0.326 +/- 0.022, 0.436 +/- 0.041 vs 0.293 +/- 0.006, 0.436 +/- 0.041 vs 0.265 +/- 0.017, respectively, P < 0.05 for NF-kappa B p65). CONCLUSION: Moxibustion repairs damaged colonic mucosa, suppresses serum IL-8, activates serum IL-10 level, and decreases expression of TLR-9 and NF-kappa B p65 in UC rats. (C) 2014 Baishideng Publishing Group Inc. All rights reserved.

Expression and signifi cance of TLR4 and HIF-1α in pancreatic ductal adenocarcinoma

AIM:To investigate the expression of toll-like receptor(TLR) 4,nuclear factor-κB(NF-κB) p65 and hypoxiainducible transcription factor 1α(HIF-1α) in pancreatic ductal adenocarcinoma and their clinical significance.METHODS:The mRNA of TLR4 and HIF-1α were investigated by real-time polymerase chain reaction in 30 cases of pancreatic ductal adenocarcinoma and its adjacent tissues,and expression of TLR4,NF-κB p65 and HIF-1α protein were detected by immunohistochemistry in 65 cases of pancreatic ductal adenocarcinoma tissues and 38 cases of corresponding adjacent tissues.The relationship between TLR4 or HIF-1α and pathologic features,as well as the association between TLR4 and HIF-1α,were also analyzed.Kaplan-Meier method was used to assess the impact of expression of TLR4 and HIF-1α on survival of patients with pancreatic cancer.RESULTS:The relative quantif ication of TLR4 and HIF-1α mRNA in tumor tissues was 0.81±0.10 and 0.87±0.11,respectively,signif icantly higher than that in adjacent tissues(0.81±0.10 vs 0.70±0.16,P=0.002;0.87±0.11 vs 0.68±0.13,P=0.000).The protein expression of TLR4,NF-κB p65 and HIF-1α in tumor tissues was 69.20%,66.15% and 70.80%,respectively,being signif icantly higher than that in adjacent normal tissues(69.20% vs 39.50%,P=0.003;66.15% vs 31.58%,P=0.001;70.80% vs 36.80%,P=0.001).There was no signif icant correlation between TLR4 or HIF-1α expression and the age,gender,tumor location,the degree of tumor differentiation in the patients(P>0.05).However,there was signif icant correlation between the expression of TLR4 or HIF-1α and tumor size,lymph node metastasis,venous invasion and clinical staging(P<0.05).The expression of TLR4 and HIF-1α had a signif icant impact on survival of patients with pancreatic adenocarcinoma.CONCLUSION:TLR4,NF-κB p65 and HIF-1α are overexpressed in pancreatic adenocarcinoma,TLR4 may be partly involved in up-regulating HIF-1α,and both synergestically promote development of pancreatic adenocarcinoma.

Sodium butyrate protects against toxin-induced acute liver failure in rats

BACKGROUND: Acute liver failure(ALF) is a serious clinical syndrome with high mortality. Sodium butyrate has been shown to alleviate organ injury in a wide variety of preclinical models of critical diseases. The aim of this study was to investigate the protective effect of sodium butyrate on ALF in rats.METHODS: All rats were randomly divided into control,model and sodium butyrate treatment groups. Except the control group, the rats were induced ALF animal model by subcutaneous injection of human serum albumin+D- galactosamine+lipopolysaccharide. After induction of ALF,the rats in the treatment group received sodium butyrate(500mg/kg) at 12-hour or 24-hour time point. Fourty-eight hours after ALF induction, the animals were sacrificed and samples were harvested. Serum endotoxin, high mobility group box-1(HMGB1), liver function parameters, tumor necrosis factoralpha(TNF-α) and interferon-gamma(IFN-γ) were measured.The expression of HMGB1 and nuclear factor-kappa B(NF-κB)p65 protein in liver tissue was detected by Western blotting. The histological changes of liver and intestine were examined. The survival duration was also observed.RESULTS: Serum endotoxin, alanine aminotransferase, HMGB1,TNF-α and IFN-γ were significantly increased and the liver histology showed more severe histopathological injury in the model group compared with the control group(P<0.05).Compared to the model group, sodium butyrate treatment significantly improved the histopathological changes in the liver and intestine, reduced serum endotoxin and inflammatory cytokines, suppressed HMGB1 and NF-кB p65 proteins in liver tissue, and prolonged the survival duration regardless of treatment at 12 hours or 24 hours after induction of ALF(P<0.05).CONCLUSIONS: Sodium butyrate protected the liver from toxin-induced ALF in rats. The mechanisms may be due to direct hepatoprotection and decreased intestinal permeability.