从NF-κB/Bcl-2信号通路调控成纤维样滑膜细胞凋亡角度探讨中医药抑制类风湿关节炎滑膜炎症机制的研究进展

类风湿关节炎(rheumatoid arthritis,RA)是一种以关节滑膜炎症为主要特征的自身免疫性疾病,成纤维样滑膜细胞(fibroblast-like synoviocytes,FLS)的抗凋亡是导致该病病情发展的主要因素。如何促进FLS凋亡并抑制炎症反应是目前RA治疗和研究的重点。核因子κB(nuclear factor kappa-B,NF-κB)/B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)信号通路在RA发病过程中发挥了关键作用,其与FLS炎症倾向、抗凋亡等密切相关。研究并探讨NF-κB/Bcl-2信号通路调控FLS凋亡的机制及作用,介绍中医药靶向该通路促进FLS凋亡进而抑制RA滑膜炎症的研究现状,旨在为中医药治疗RA的研究提供一定基础。

红景天苷调控NF-κB、Bcl-2信号通路对烟雾暴露大鼠氧化应激及肺血管内皮细胞凋亡的影响

目的探究红景天苷(SDS)通过调控B淋巴细胞瘤-2(Bcl-2)、核转录因子-κB(NF-κB)信号通路对烟雾暴露大鼠氧化应激及肺血管内皮细胞凋亡的机制。方法选取60只大鼠分为对照组、模型组、SDS 10 mg/kg、SDS 40 mg/kg、SDS 70 mg/kg、醋酸泼尼松(PNS)组,每组10只,除对照组外,其余均建立烟雾致肺损伤模型,对照组与模型组大鼠每日采用等量生理盐水灌胃;SDS高、中、低剂量组分别按70、40、10 mg/kg剂量给予SDS混悬液2 mL灌胃;PNS组给予PNS混悬液3.6 mg/kg灌胃,均每日1次,连续21 d。结果与对照组比较,模型组大鼠血清中丙二醛(MDA)、Bcl-2相关X因子(Bax)、NF-κB、p-NF-κB及细胞凋亡升高,超氧化物歧化酶(SOD)、Bcl-2降低(P<0.05);与模型组比较,SDS各剂量组MDA、Bax、NF-κB、p-NF-κB及细胞凋亡均有所降低,SOD、Bcl-2也有所升高(P<0.05),且模型组与SDS 10 mg/kg组比较无明显差异(P>0.05),SDS 40 mg/kg组与PNS组比较无明显差异(P>0.05)。对照组大鼠肺组织良好;模型组肺损伤严重;药物干预后各组肺损伤均减轻。结论SDS可改善烟雾暴露大鼠肺损伤、氧化应激及血管内皮细胞凋亡,其机制可能与调控Bax/Bcl-2及NF-κB相关。

Kuicolong-yu enema decoction retains traditional Chinese medicine enema attenuates inflammatory response ulcerative colitis through TLR4/NF-κB signaling pathway

BACKGROUND Ulcer colitis(UC)is a chronic,nonspecific,and noninfectious inflammatory bowel disease.Recently,Toll-like receptors(TLRs)have been found to be closely associated with clinical inflammatory diseases.Achieving complete remission in patients with intermittent periods of activity followed by dormancy is challenging.Moreover,no study has explored the mechanism by which Kuicolong-yu enema decoction retains traditional Chinese medicine enemas to attenuate the inflammatory response in UC.AIM To explore the mechanism by which Kuicolong-yu enema decoction retains traditional Chinese medicine enemas to attenuate the inflammatory response in UC.METHODS This prospective clinical study included patients who met the exclusion criteria in 2020 and 2021.The patients with UC were divided into two groups(control and experimental).The peripheral blood of the experimental and control groups were collected under aseptic conditions.The expression of TLR4 protein,NF-κB,IL-6,and IL-17 was detected in the peripheral blood of patients in the experimental group and control group before and 1 month after taking the drug.Linear co rrelation analysis was used to analyze the relationship between the expression level of TLR4 protein and the expression levels of downstream signal NF-κB and inflammatory factors IL-6 and IL-17,and P<0.05 was considered statistically significant.RESULTS There were no significant differences in the patient characteristics between the control and experimental groups.The results showed that the expression levels of TLR4 and NF-κB in the experimental group were significantly lower than those in the control group(P<0.05).The levels of IL-6 and IL-17 in the experimental group were significantly lower than those in the control group(P<0.05).The TLR4 protein expression in the experimental group was positively correlated with the expression level of downstream signal NF-κB and was positively correlated with the levels of downstream inflammatory cytokines IL-6 and IL-17(r=0.823,P<0.05).CONCLUSION Kuicolong-yu enema decoction retains traditional Chinese medicine enema attenuates the inflammatory response of UC through the TLR4/NF-κB signaling pathway.

Calcitriol attenuates liver fibrosis through hepatitis C virus nonstructural protein 3-transactivated protein 1-mediated TGF β1/Smad3 and NF-κB signaling pathways

BACKGROUND Hepatic fibrosis is a serious condition,and the development of hepatic fibrosis can lead to a series of complications.However,the pathogenesis of hepatic fibrosis remains unclear,and effective therapy options are still lacking.Our group identified hepatitis C virus nonstructural protein 3-transactivated protein 1(NS3TP1) by suppressive subtractive hybridization and bioinformatics analysis,but its role in diseases including hepatic fibrosis remains undefined.Therefore,additional studies on the function of NS3TP1 in hepatic fibrosis are urgently needed to provide new targets for treatment.AIM To elucidate the mechanism of NS3TP1 in hepatic fibrosis and the regulatory effects of calcitriol on NS3TP1.METHODS Twenty-four male C57BL/6 mice were randomized and separated into three groups,comprising the normal,fibrosis,and calcitriol treatment groups,and liver fibrosis was modeled by carbon tetrachloride(CCl4).To evaluate the level of hepatic fibrosis in every group,serological and pathological examinations of the liver were conducted.TGF-β1 was administered to boost the in vitro cultivation of LX-2 cells.NS3TP1,α-smooth muscle actin(α-SMA),collagen I,and collagen Ⅲ in every group were examined using a Western blot and real-time quantitative polymerase chain reaction.The activity of the transforming growth factor beta 1(TGFβ1)/Smad3 and NF-κB signaling pathways in each group of cells transfected with pcDNA-NS3TP1 or siRNA-NS3TP1 was detected.The statistical analysis of the data was performed using the Student’s t test.RESULTS NS3TP1 promoted the activation,proliferation,and differentiation of hepatic stellate cells(HSCs)and enhanced hepatic fibrosis via the TGFβ1/Smad3 and NF-κB signaling pathways,as evidenced by the presence of α-SMA,collagen I,collagen Ⅲ,p-smad3,and p-p65 in LX-2 cells,which were upregulated after NS3TP1 overexpression and downregulated after NS3TP1 interference.The proliferation of HSCs was lowered after NS3TP1 interference and elevated after NS3TP1 overexpression,as shown by the luciferase assay.NS3TP1 inhibited the apoptosis of HSCs.Moreover,both Smad3 and p65 could bind to NS3TP1,and p65 increased the promoter activity of NS3TP1,while NS3TP1 increased the promoter activity of TGFβ1 receptor I,as indicated by coimmunoprecipitation and luciferase assay results.Both in vivo and in vitro,treatment with calcitriol dramatically reduced the expression of NS3TP1.Calcitriol therapy-controlled HSCs activation,proliferation,and differentiation and substantially suppressed CCl4-induced hepatic fibrosis in mice.Furthermore,calcitriol modulated the activities of the above signaling pathways via downregulation of NS3TP1.CONCLUSION Our results suggest that calcitriol may be employed as an adjuvant therapy for hepatic fibrosis and that NS3TP1 is a unique,prospective therapeutic target in hepatic fibrosis.

Acupuncture at Back-Shu point improves insomnia by reducing inflammation and inhibiting the ERK/NF-κB signaling pathway

BACKGROUND Insomnia is a disease where individuals cannot maintain a steady and stable sleep state or fail to fall asleep.Western medicine mainly uses sedatives and hypnotic drugs to treat insomnia,and long-term use is prone to drug resistance and other adverse reactions.Acupuncture has a good curative effect and unique advantages in the treatment of insomnia.AIM To explore the molecular mechanism of acupuncture at Back-Shu point for the treatment of insomnia.METHODS We first prepared a rat model of insomnia,and then carried out acupuncture for 7 consecutive days.After treatment,the sleep time and general behavior of the rats were determined.The Morris water maze test was used to assess the learning ability and spatial memory ability of the rats.The expression levels of inflammatory cytokines in serum and the hippocampus were detected by ELISA.qRTPCR was used to detect the mRNA expression changes in the ERK/NF-κB signaling pathway.Western blot and immunohistochemistry were carried out to evaluate the protein expression levels of RAF-1,MEK-2,ERK1/2 and NF-κB.RESULTS Acupuncture can prolong sleep duration,and improve mental state,activity,diet volume,learning ability and spatial memory.In addition,acupuncture increased the release of 1L-1β,1L-6 and TNF-αin serum and the hippocampus and inhibited the mRNA and protein expression of the ERK/NF-κB signaling pathway.CONCLUSION These findings suggest that acupuncture at Back-Shu point can inhibit the ERK/NF-κB signaling pathway and treat insomnia by increasing the release of inflammatory cytokines in the hippocampus.

Involvement of ayu NOD2 in NF-κB and MAPK signaling pathways: Insights into functional conservation of NOD2 in antibacterial innate immunity

Nucleotide oligomerization domain 2(NOD2) is a major cytoplasmic sensor for pathogens and is critical for the clearance of cytosolic bacteria in mammals.However, studies regarding NOD2, especially the initiated signaling pathways, are scarce in teleost species. In this study, we identified a NOD2 molecule(PaNOD2) from ayu(Plecoglossus altivelis).Bioinformatics analysis showed the structure of NOD2 to be highly conserved during vertebrate evolution. Dual-luciferase reporter assays examined the activation of NF-κB signaling and Western blotting analysis detected the phosphorylation of three MAP kinases(p-38, Erk1/2, and JNK1/2).Functional study revealed that, like its mammalian counterparts, PaNOD2 was the receptor of the bacterial cell wall component muramyl dipeptide(MDP), and the leucine-rich repeat motif was responsible for the recognition and binding of Pa NOD2 with the ligand. Overexpression of PaNOD2 activated the NF-κB signaling pathway, leading to the upregulation of inflammatory cytokines, including TNF-α and IL-1β in HEK293 T cells and ayu head kidney-derived monocytes/macrophages(MO/MΦ).Particularly, we found that PaNOD2 activated the MAPK signaling pathways, as indicated by the increased phosphorylation of p-38, Erk1/2, and JNK1/2, which have not been characterized in any teleost species previously. Our findings proved that the NOD2 molecule and initiated pathways are conserved between mammals and ayu. Therefore, ayu could be used as an animal model to investigate NOD2-based diseases and therapeutic applications.

β-arrestin 2 attenuates lipopolysaccharide-induced liver injury via inhibition of TLR4/NF-κB signaling pathwaymediated inflammation in mice

AIM To study the role and the possible mechanism of β-arrestin 2 in lipopolysaccharide(LPS)-induced liver injury in vivo and in vitro.METHODS Male β-arrestin 2^(+/+) and β-arrestin 2^(-/-)C57 BL/6 J mice were used for in vivo experiments, and the mouse macrophage cell line RAW264.7 was used for in vitro experiments. The animal model was established via intraperitoneal injection of LPS or physiological sodium chloride solution. Blood samples and liver tissues were collected to analyze liver injury and levels of pro-inflammatory cytokines. Cultured cell extracts were collected to analyze the production of pro-inflammatory cytokines and expression of key molecules involved in the TLR4/NF-κB signaling pathway.RESULTS Compared with wild-type mice, the β-arrestin 2 knockout mice displayed more severe LPS-induced liver injury and significantly higher levels of proinflammatory cytokines, including interleukin(IL)-1β, IL-6, tumor necrosis factor(TNF)-α, and IL-10. Compared with the control group, pro-inflammatory cytokines(including IL-1β, IL-6, TNF-α, and IL-10) produced by RAW264.7 cells in the β-arrestin 2 si RNA group were significantly increased at 6 h after treatment with LPS. Further, key molecules involved in the TLR4/NF-κB signaling pathway, including phosphoIκBα and phosho-p65, were upregulated.CONCLUSION β-arrestin 2 can protect liver tissue from LPS-induced injury via inhibition of TLR4/NF-κB signaling pathwaymediated inflammation.

CARD11与NF-_κB及Bcl-2在弥漫性大B细胞淋巴瘤中的表达及意义

目的探讨CARD11与NF-κB及Bcl-2在弥漫性大B细胞淋巴瘤(DLBCL)不同免疫分型中的表达情况和相互作用关系,了解3种分子标记物对DLBCL的发展过程和临床特征的影响。方法收集临床资料,选取DL-BCL标本70例,反应性增生的淋巴结20例。应用免疫组织化学染色法和组织芯片技术对DLBCL进行免疫分型并检测分型中CARD11、NF-κB和Bcl-2的表达情况。对收集到的数据资料进行整理和统计学分析。结果 70例DLBCL组织标本中,GCB30例,non-GCB40例;Maxvision免疫组化结果显示在DLBCL组织标本70例中,CARD11阳性42例,阳性率60.0%;反应增生性淋巴结中CARD11阳性5例,阳性率25.0%。结论在DLBCL亚型的分类中CARD11、NF-κB和Bcl-2具有重要的作用,为临床治疗和预防提供了重要参考。

Phycocyanin attenuates X-ray-induced pulmonary inflammation via the TLR2-MyD88-NF-κB signaling pathway

Phycocyanin (PC), a natural algal protein, is reported for having anti-oxidant and antiinfl ammatory properties. We investigated its ability to attenuate lung infl ammation in mice subjected to X-ray radiation. Male C57BL/6 mice were assigned to the control, total body irradiation, PC pretreatment, and PC treatment groups. Mice in the PC pretreatment group were gavaged with 200 mg/kg PC for 7 consecutive days before irradiation, and those in the PC treatment group were gavaged with 200 mg/kg PC for 7 consecutive days after irradiation. Lungs were collected on Day 7 after irradiation exposure. Hematoxylin and eosin staining of mouse lung sections showed considerable infl ammation damage 7 days after irradiation compared with the control lung but a reduction in pathological injury in the PC treatment group. Pretreatment or treatment with PC signifi cantly decreased levels of interleukin-6 and tumor necrosis factor-α in the lung, and also increased the relative mRNA expression of superoxide dismutase and glutathione. In vivo, PC signifi cantly reduced the expression of Toll-like receptor TLR2, myeloid diff erentiation primary response Myd88, and nuclear factor NF-κB, at both the transcriptional and translation level. Taken together, these data indicated that PC attenuated lung infl ammatory damage induced by radiation by blocking the TLR2- MyD88-NF-κB signaling pathway. Therefore, PC could be a protective agent against radiation-induced infl ammatory damage in normal tissues.

Immunoregulatory polysaccharides from Apocynum venetum L.flowers stimulate phagocytosis and cytokine expression via activating the NF-κB/MAPK signaling pathways in RAW264.7 cells

Two immunomodulatory polysaccharides(Vp2a-Ⅱ and Vp3) were isolated and identified from Apocynum venetum L. flowers, and their innate immune-stimulating functions and working mechanisms were evaluated in RAW264.7 cells. Both the level of released nitric oxide(NO) and expression of inducible nitric oxide synthase(iNOS) m RNA were significantly enhanced in the RAW264.7 macrophages cells treated by Vp2a-Ⅱ and Vp3. Vp2a-Ⅱ(100–800 μg/m L) and Vp3(400 μg/mL) could significantly increase the phagocytic activity of RAW264.7 cells and the secretion and m RNA expression of TNF-α and IL-6 in a concentrationdependent manner through affecting mitogen-activated protein kinase(MAPK) activity and nuclear factor κB(NF-κB) nuclear translocation. Vp2a-Ⅱ might activate the MAPK signaling pathways and induce the nuclear translocation of NF-κB p65, whilst Vp3 likely activated the NF-κB and MAPK signaling pathways without influencing the p38 MAPK route.

NF-κB、bcl-2、bax在胃不典型增生、胃癌中的表达及其意义

目的:探讨核因子κB(nuclear factor kappa B,NFκ-B)、bcl-2及bax在胃不典型增生及胃癌中的表达及在胃癌发生中的作用。方法:采用免疫组化Power Vision两步法检测84例胃癌和54例胃不典型增生组织中NFκ-B、bcl-2及bax的表达,TNNEL法染色检测凋亡指标的改变。结果:NFκ-B、bcl-2、bax在84例胃癌组织中的阳性表达率分别为59.52%、52.38%、16.67%,在33例胃轻度不典型增生组织中阳性表达率分别为24.24%、21.21%、33.33%,三者在胃癌与轻度不典型增生组之间差异有统计学意义(P<0.05)。NFκ-B、bcl-2表达与肿瘤的大小相关(P<0.05),NFκ-B与bcl-2之间呈正相关(P<0.05)。在轻度不典型增生、重度不典型增生与胃癌组织中AI分别为(1.29±0.50)%、(0.96±0.36)%、(0.70±0.43)%,轻度不典型增生与重度不典型增生、胃癌之间差异均有统计学意义(P<0.05)。结论:在胃癌中NFκ-B可通过上调bcl-2的表达、改变bcl-2与bax的比值而抑制肿瘤细胞凋亡,进而影响肿瘤的生长、浸润、转移。

环烯醚萜苷对大鼠脑梗死后NF-κB与凋亡调节因子Bcl-2/Bax表达变化的影响

目的观察光化学法诱导大鼠脑梗死形成过程中核转录因子(Nuc lear factor-kappa B,NF-κB)与凋亡调节因子Bax和Bc l-2表达的变化,并探讨山茱萸环烯醚萜苷(iridoid gly-coside,IG)对这一过程的影响。方法大鼠预先灌胃给药7d后,制作光化学致脑梗死模型,采用免疫组织化学法检测脑组织中NF-κB表达的变化,W estern b lot技术检测凋亡调节因子Bax和Bc l-2表达的变化。结果模型组与对照组相比,脑皮层内NF-κB和Bax蛋白表达增多,Bc l-2蛋白表达减少。与模型组相比,IG能够明显的减少NF-κB和Bax蛋白表达,增高Bc l-2蛋白的表达。结论IG可通过影响脑内NF-κB和凋亡调节基因的表达而发挥对脑梗死的治疗作用。

青蒿琥酯对佐剂性关节炎大鼠踝关节滑膜的NF-κB、bcl-2表达的影响

目的:观察青蒿琥酯对佐剂性关节炎大鼠踝关节滑膜的病理形态和滑膜细胞的NF-κB、bcl-2表达的影响。方法:42只♂性Wistar大鼠随机分为5组,空白组(8只)、模型对照组(8只)、青蒿琥酯Ⅰ组(8只,40mg.kg-1)、青蒿琥酯Ⅱ组(8只,20mg.kg-1)、青蒿琥酯加甲氨蝶呤组(10只,青蒿琥酯20mg.kg-1、甲氨蝶呤每3d0.5mg.kg-1)。采用大鼠右后足跖皮内注射完全性福氏佐剂0.1mL,致炎形成佐剂性关节炎(AA)模型,于造模12d后开始给药。给药12d后处死动物,组织切片HE染色观察大鼠踝关节滑膜的组织形态学变化,免疫组化ABC法检测滑膜组织中核转录因子NF-κB、凋亡相关蛋白bcl-2的蛋白表达水平。结果:青蒿琥酯能明显减轻滑膜细胞的增生,减少滑膜层和滑膜下层淋巴细胞浸润。青蒿琥酯Ⅰ组、青蒿琥酯Ⅱ组及青蒿琥酯加甲氨蝶呤组关节滑膜细胞的bcl-2和NF-κB表达明显低于模型组,差异有显著性(P<0.05)。结论:青蒿琥酯可显著改善佐剂性关节炎大鼠的滑膜病变,降低滑膜细胞的bcl-2和NF-κB的蛋白表达。

胃肠道间质瘤中PTEN、蛋白激酶B与转录因子家族NF-κB、bcl-2关系的研究

目的研究磷酸化抑癌基因(PTEN)、蛋白激酶B(Akt)基因与转录因子家族NF-κB、B细胞淋巴瘤/白血病2(bcl-2)在胃肠道间质瘤(GIST)中的表达及意义。方法应用RT-PCR及Westernblot检测124例GIST中Akt、PTEN、NF-κB、bcl-2基因mRNA和蛋白表达情况。结果在GIST中,随着NIH分级增高,Akt的mRNA及蛋白表达水平上调,而抑癌基因PTEN的mRNA及蛋白表达水平下调,二者阳性表达率与肿瘤侵袭转移和黏膜受侵关系密切,PTEN与pAkt蛋白阳性表达之间呈显著负相关(r=-0.386,P=0.024)。NF-κB、bcl-2的mRNA的表达变化与NIH分级以及肿瘤侵袭转移、黏膜受侵密切相关,而与年龄、性别、组织学类型无关;NF-κB和bcl-2mRNA表达水平呈正相关;Akt和NF-κB、bcl-2的mRNA表达水平呈正相关。结论 GIST中PTEN基因失活可以活化蛋白激酶Akt,进一步激活NF-κB转录因子,上调凋亡相关因子bcl-2mRNA水平的表达。

高三尖杉酯碱对K562细胞NF-κB和BCL-2蛋白表达的影响

本研究旨在探讨高三尖杉酯碱(HHT)对K562细胞增殖、凋亡及BCL-2和NF-κB蛋白表达的影响。不同浓度HHT作用K562细胞后用MTT法、流式细胞仪、Western blot等方法分别检测细胞增殖、凋亡、BCL-2及NF-κB蛋白表达水平。结果表明,HHT作用48 h,浓度依赖性抑制K562细胞增殖,IC50为43.89 ng/ml。HHT 10 ng/ml作用48 h,K562细胞凋亡率明显增高;细胞周期阻滞于G0/G1期;BCL-2和NF-κB蛋白表达水平明显低于对照组(P<0.05)。结论:HHT对K562细胞有显著的增殖抑制、细胞周期阻滞和凋亡诱导作用,抑制NF-κB和BCL-2表达可能是其抗CML的机制之一。

NF-κB活性变化对HL-60细胞凋亡及WT1、Bcl-2基因表达的影响

目的为探讨NF-κB变化对白血病细胞凋亡的影响及其机制。方法用PDTC抑制HL-60细胞NF-κB的活性,观察NF-κB活性抑制前后阿糖胞苷诱导的HL-60细胞凋亡的变化及WT1、Bcl-2表达水平的变化。结果NF-κB活性抑制能明显增强阿糖胞苷诱导的HL-60细胞凋亡作用,同时明显下调WT1、Bcl-2的表达。结论NF-κB活性抑制使白血病细胞凋亡增加,其作用可通过直接下调WT1的表达和间接下调Bcl-2的表达来实现.

Study on the Hepatoprotective Effect of Oxalis coriniculata L. and Related Mechanism by Regulating Oxidative Stress and TLR-2/NF-κB Signaling Pathway

[Objectives]This study aimed to explore the protective effect of Oxalis coriniculata L.on rats with acute liver injury induced by carbon tetrachloride(CCl4)and related mechanism by regulating oxidative stress and the TLR-2 TLR-2/NF-κB signaling pathway.[Methods]A total of 48 female rats were randomly and evenly divided into normal group,model group,silymarin group(0.12 g/kg),and high(16 g/kg),middle(8 g/kg)and low-dose(4 g/kg)O.coriniculata L.groups.The rats in the groups were intragastrically administered with 5 mL/kg of corresponding drugs(equal-volume distilled water for normal group and control group),respectively.The administration was conducted twice a day,for 10 consecutive days.After 2 h of the last administration,the rats in all the groups except the normal group were intraperitoneally injected with 12%carbon tetrachloride(CCl4)olive oil solution(5 mL/kg),respectively to establish liver injury rat models.After 16 h,the eyeball blood of the rats was collected,and their liver tissues were collected for preparation of HE sections.The biochemical indicators detected included aspartate aminotransferase(AST),alanine aminotransferase(ALT),total superoxide dismutase(T-SOD)and glutathione peroxidase(GSH-Px)activity and malondialdehyde(MDA)content in the serum.The contents of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6)in the serum were detected by ELISA.The expression of Toll-like receptor-2(TLR-2)and nuclear factor-κB(NF-κB)in liver tissue was detected using Western blotting.The pathological changes of liver were observed under light microscope.[Results]Compared with the normal group,the ALT,AST activity and MDA,IL-1β,IL-6,TNF-αlevels in rat serum significantly increased(P<0.01),the GSH-Px,T-SOD activity in rat serum significantly decreased(P<0.01),and the expression of TLR-2 and NF-κB in liver tissue was up-regulated(P<0.01)in the model group.Compared with the model group,the ALT,AST activity and MDA,IL-1β,IL-6 and TNF-αlevels in rat serum reduced(P<0.05,P<0.01),the GSH-Px and T-SOD activity in rat serum increased(P<0.05,P<0.01),and the expression of TLR-2 and NF-κB in liver tissue was down-regulated(P<0.05,P<0.01)in the O.coriniculata L.administration groups.Pathological sections show that O.coriniculata L.had an improving effect on rats with acute liver injury induced by CCl4.[Conclusions]O.coriniculata L.has a good protective effect on rats with acute liver injury induced by CCl4.Its mechanism may be related to inhibition of oxidative stress,inhibition of inflammatory response and regulation of the TLR-2/NF-κB signaling pathway.

BARF1基因对鼻咽癌细胞NF-κB信号通路及bcl-2基因表达的影响

目的:探讨EB病毒编码的BARF1基因对鼻咽癌细胞NF-κB信号通路活性及bcl-2基因表达的影响。方法:以pSG5空载体转染鼻咽癌细胞(CNE1-SG)为对照,分别采用荧光抗体染色法和蛋白质印迹法检测稳定表达BARF1的鼻咽癌细胞系CNE1(CNE1-BARF1)中NF-κB蛋白的核转位和Bcl-2蛋白的表达变化。结果:与CNE1-SG相比,CNE1-BARF1细胞中发生明显的NF-κB核转位,同时Bcl-2蛋白表达量显著增加(t=3.89,P<0.05)。采用NF-κB的特异性抑制剂Bay11-7082处理后,NF-κB蛋白的核转位明显减少,同时Bcl-2蛋白表达量显著下降(t=3.13,P<0.05)。结论:BARF1基因通过激活NF-κB信号通路上调鼻咽癌细胞中bcl-2基因的表达。

SAA蛋白与BCL-2、Caspase-3及NF-κB关系的研究

目的:探讨血清淀粉样蛋白A(SAA)的超表达和抑制表达对BCL-2、Caspase-3及NF-κB这3种信号分子蛋白表达的影响。方法:将课题组前期已构建成功的pc DNA3.1(+)-SAA超表达载体和p GPU6/GFP/Neo-SAA表达干扰载体采用脂质体法分别转染鼻咽癌CNE2细胞,建立SAA蛋白超表达的pc DNA3.1(+)-SAA-CNE2细胞系和干扰SAA蛋白表达的p GPU6/GFP/Neo-SAA-CNE2细胞系。pc DNA3.1(+)-SAACNE2细胞、CNE2细胞、p GPU6/GFP/Neo-SAA-CNE2细胞中SAA蛋白的表达量会逐渐减少,当SAA蛋白的表达量改变时,通过免疫印迹法检测BCL-2、Caspase-3及NF-κB在pc DNA3.1(+)-SAA-CNE2细胞、CNE2细胞、p GPU6/GFP/Neo-SAA-CNE2细胞中的表达。结果:SAA蛋白的表达在pc DNA3.1(+)-SAA-CNE2细胞系中明显增多(P<0.05),在p GPU6/GFP/Neo-SAA-CNE2细胞系中显著减少(P<0.05),由此可知,pc DNA3.1(+)-SAA超表达载体和p GPU6/GFP/Neo-SAA表达干扰载体成功转染CEN2细胞。免疫印迹检测结果示:随着SAA蛋白表达的增多,BCL-2的表达增多(P<0.05)、Caspase-3及NF-κB的表达减少(P<0.05)。结论:SAA蛋白在人鼻咽癌CNE2细胞中的表达与抑制凋亡的BCL-2蛋白表达有协同作用,对凋亡调节分子Caspase-3及NF-κB蛋白的表达有抑制作用,可推测SAA蛋白可能有促进肿瘤生长、转移的作用。

肝癌组织中Bcl-2和NF-κB的表达与AFP相关性的研究

目的:观察凋亡相关基因Bcl-2、核因子NF-κB在肝细胞性肝癌(HCC)组织中的表达,并探讨二者与AFP之间的关系。方法:采用免疫组织化学方法检测40例HCC组织(其中AFP阳性20例,AFP阴性20例)和10例良性肿瘤组织中Bcl-2、NF-κB的表达。结果:40例HCC患者Bcl-2阳性表达率为45.0%,NF-κB阳性表达率为57.5%;其中AFP阳性细胞Bcl-2表达率为55.0%,AFP阴性细胞为35.0%;AFP阳性细胞NF-κB表达率为75.0%,AFP阴性细胞为40.0%。HCC细胞NF-kB和Bcl-2的表达呈正相关(r=0.778,P<0.01)。结论:Bcl-2、NF-κB的异常表达同HCC的抗凋亡和耐药性有一定关系,AFP阳性HCC细胞的Bcl-2、NF-κB阳性表达率更高。Bcl-2和NF-κB可以考虑作为预后的判断指标之一。