基于BDNF通路探讨化痰通络汤治疗卒中后认知功能障碍的研究

目的 观察化痰通络汤治疗卒中后认知功能障碍(PSCI)痰瘀阻络证的临床疗效及其对患者血清脑源性神经营养因子(BDNF)通路相关因子的影响。方法 收集符合纳入标准的患者60例,随机分为对照组和治疗组各30例。对照组予基础治疗和尼莫地平治疗,治疗组在对照组治疗基础上加服化痰通络汤,2组疗程均为4周。治疗前后比较2组简易精神状态评价量表(MMSE)、中医证候积分及血清BDNF、核转录因子κB(NF-κB)、B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)水平变化,治疗后评估2组患者中医临床疗效,治疗期间观察2组患者不良反应发生情况。结果 治疗后,2组患者MMSE评分显著增加,中医证候积分总积分明显降低(P<0.01),治疗组优于对照组(P<0.01);2组患者血清BDNF、NF-κB、Bcl-2水平明显升高(P<0.05,P<0.01),Bax水平明显下降(P<0.01),治疗组优于对照组(P<0.01)。结论 化痰通络汤能够改善痰瘀阻络型PSCI患者临床症状,安全有效,其疗效机制可能与调控BDNF通路,抑制神经细胞凋亡有关。

Helicobacter pylori tumor necrosis factor-α inducing protein promotes cytokine expression via nuclear factor-κB

AIM:To study the effects of Helicobacter pylori(H. pylori)tumor necrosis factor-α(TNF)inducing protein (Tip-α)on cytokine expression and its mechanism. METHODS:We cloned Tip-αfrom the H.pylori strain 26695,transformed Escherichia coli with an expression plasmid,and then confirmed the expression product by Western blotting.Using different concentrations of Tip-αthat affected SGC7901 and GES-1 cells at different times,we assessed cytokine levels using enzyme-linked immunosorbent assay.We blocked SGC7901 cells with pyrrolidine dithiocarbamate(PDTC),a specific inhibitor of nuclear factorκB(NF-κB).We then detected interleukin(IL)-1βand TNF-αlevels in SGC7901 cells. RESULTS:Western blot analysis using an anti-Tip-α antibody revealed a 23-kDa protein,which indicated that recombinant Tip-αprotein was recombined successfully.The levels of IL-1β,IL-8 and TNF-αwere sig-nificantly higher following Tip-αinterference,whether GES-1 cells or SGC-7901 cells were used(P<0.05).However,the levels of cytokines(including IL-1β,IL-8 and TNF-α)secreted by SGC-7901 cells were greater than those secreted by GES-1 cells following treatment with Tip-αat the same concentration and for the same duration(P<0.05).After blocking NF-κB with PDTC, the cells(GES-1 cells and SGC-7901 cells)underwent interference with Tip-α.We found that IL-1βand TNF-αlevels were significantly decreased compared to cells that only underwent Tip-αinterference(P<0.05). CONCLUSION:Tip-αplays an important role in cyto-kine expression through NF-κB.

Effects of Ginsenoside Rg1 on nuclear factor-kappa B activity in beta amyloid protein-treated neural cells

BACKGROUND： Modern pharmacological studies have shown that Ginsenoside Rgl is one of the active components of ginseng that promote intelligence in the nervous system. Ginsenoside Rgl can improve memory and learning in mouse models of β-amyloid protein （Aβ）-induced dementia. OBJECTIVE： To investigate whether effects of Ginsenoside Rgl against Aβ are associated with activity of nuclear factor-kappa B （NF-κB）. DESIGN, TIME AND SETTING： The randomized performed at the DME Center, Institute of Clinica controlled, cell biological experiment was Pharmacology, Guangzhou University of Chinese Medicine, China from July 2005 to May 2006. MATERIALS： Beta-amyloid fragment 25-35 （Aβ25-35） was supplied by the Neural Biochemical Laboratory, Xuanwu Hospital, Capital Medical University, China. Ginsenoside Rgl was obtained from National Institute for the Control of Pharmaceutical and Biological Products, China. Rabbit anti-rat NF-κB p65 antibody was purchased from Santa Cruz Biotechnology, USA. METHODS： Hippocampal neurons and cortical astrocytes of neonatal Sprague Dawley rats were harvested and treated with various concentrations （0, 5, 10, 20, and 40 μmol/L） of Aβ for 6, 12, and 24 hours to establish cellular models of Alzheimer＇s disease. Cellular models were pretreated with various concentrations of Ginsenoside Rgl （1,2, 4, 8, and 16 μmol/L）. According to cell morphology and activity, the following conditions were selected： 40 μmol/L Aβ for 24 hours, as well as 2, 4, and 8 μmol/L Ginsenoside Rg1. NF-κB activity was observed using immunofluorescence and cytochemical staining. MAIN OUTCOME MEASURES： Morphology and viability of hippocampal neurons and cortical astrocytes, and activities of NF-κB were measured. RESULTS： Hippocampal neuron activity was significantly greater in the normal and 2 and 4 μmol/L Ginsenoside Rgl groups compared with the model group （P 〈 0.05）. Astrocyte activity was significantly greater in the normal, 1,2, 4, 8, and 16 μmol/L Ginsenoside Rgl groups compared with the model group （P 〈 0.05）. NF-κB activity of hippocampal neurons was significantly greater in the normal, 2, 4, and 8 μmol/L Ginsenoside Rgl groups compared with the model group （P 〈 0.01）. NF-κB activity of astrocytes was significantly less in the normal, 2, 4, and 8 μmol/L Ginsenoside Rgl groups compared with the model group （P 〈 0.01 or P 〈 0.05）. No significant difference in NF-κB activity was determined between the 2 μmol/L Ginsenoside Rgl and normal groups （P 〉 0.05）. CONCLUSION： Ginsenoside Rgl protected neural cells by upregulating NF-κB activity in neurons and downregulating NF-κB activity in astrocytes. Ginsenoside Rgl （2 μmol/L） maintained cell activity and NF-κB activity at normal levels.

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.

An Experimental Study on the Role of Nuclear Factor-κB in the Signal Conduction of Protein Kinase C Regulating the Proliferation and Apoptosis of T Lymphocytes in Asthma

To explore the role of nuclear factor-κB(NF-κB) in the signal pathway of protein kinase C (PKC) regulating the proliferation and apoptosis of T lymphocytes in asthma. T lymphocytes were isolated from the asthmatic model of guinea pigs and the asthmatic patients. Either the T cells stimulated with PMA alone or those stimulated with PMA together with pyrrolidine dithiocarbamate (PDTC) were incubated for 1 and 24?h. The proliferation of and the presence of NF-κB in the cells incubated for 1?h were observed by MTT and immunohistochemical staining, respectively. And the cells incubated for 24?h were observed for the apoptosis by TUNEL. All the assays were paralleled with controls, and all the data were analyzedstatistically with the software SAS. The percentage of cells of nuclear positive staining of NF-κB and the proliferation of T lymphocytes from asthmatic guinea pigs and asthmatic patients stimulated with PMA were significantly higher than those of T lymphocytes from asthmatic guinea pigs and asthmatic patients stimulated without PMA respectively (P<0.01) and those of T lymphocytes from normal control guinea pigs and normal control persons stimulated with PMA respectively (P<0.01), and were significantly reduced by PDTC (P<0.01). The apoptosis index of T lymphocytes from asthmatic guinea pigs and asthmatic patients stimulated with PMA were significantly lower than those of T lymphocytes from asthmatic guinea pigs and asthmatic patients stimulated without PMA respectively (P<0.01) and those of T lymphocytes from normal control guinea pigs and normal control persons stimulated with PMA respectively (P<0.01), and were significantly induced by PDTC (P<0.01). There were good positive correlation between the percentage of cells of nuclear staining of NF-κB of T lymphocytes and the proliferation of T lymphocytes (r=0.51-0.72, P<0.001), and also good negative correlation between the percentage of cells of nuclear staining of NF-κB and the apoptosis index of T lymphocytes (r=-0.55-0.71, P<0.001, respectively). It concludes that the active PKC of asthmatic T lymphocytes promoting the proliferation and inhibiting the apoptosis of T lymphocytes may be mediated by activating NF-κB, the activation of PKC-NF-κB signal pathway of T lymphocytes NF-κB may play an important role in the pathogenesis of asthma.

山姜素调控PI3K/Akt/NF-κB信号通路对冠心病大鼠心肌损伤的影响

目的:探讨山姜素调控磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/核转录因子-κB(NF-κB)信号通路对冠心病(CHD)大鼠心肌损伤的影响。方法:选取健康成年SPF级SD雄性大鼠60只,随机分为空白组、模型组、山姜素小剂量组、山姜素中剂量组、山姜素高剂量组各12只,采用Western blot法检测大鼠PI3K/Akt/NF-κB信号通路蛋白表达,采用ELISA法检测大鼠炎症因子指标[肿瘤坏死因子α(TNF-α)、白介素-1β(IL-1β)、白介素-18(IL-18)]和氧化应激指标[活性氧(ROS)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)],观察并记录大鼠心肌损伤情况。结果:与空白组比较,模型组的心肌缺血和心肌梗死面积更大(P<0.05),与模型组比较,山姜素小、中、高剂量组的心肌缺血和心肌梗死面积均缩小(P<0.05),且高剂量组的心肌缺血和心肌梗死面积小于山姜素中、小剂量组,中剂量组心肌缺血和心肌梗死面积小于山姜素小剂量组(P<0.05)。与空白组比较,模型组的PI3K、Akt、NF-κB更高(P<0.05),与模型组比较,山姜素小、中、高剂量组的PI3K、Akt、NF-κB均升高(P<0.05),且高剂量组的PI3K、Akt、NF-κB高于山姜素中、小剂量组,中剂量组PI3K、Akt、NF-κB高于山姜素小剂量组(P<0.05)。与空白组比较,模型组的TNF-α、IL-1β、IL-18更高(P<0.05),与模型组比较,山姜素小、中、高剂量组的TNF-α、IL-1β、IL-18均降低(P<0.05),且山姜素高剂量组的TNF-α、IL-1β、IL-18低于中、小剂量组,山姜素中剂量组的TNF-α、IL-1β、IL-18低于山姜素小剂量组(P<0.05)。与空白组比较,模型组的ROS、GSH-Px、MDA更高(P<0.05),与模型组比较,山姜素小、中、高剂量组的ROS、GSH-Px、MDA均降低(P<0.05),且山姜素高剂量组ROS、GSH-Px、MDA低于山姜素中、小剂量组,山姜素中剂量组的ROS、GSH-Px、MDA低于山姜素小剂量组(P<0.05)。结论:山姜素可通过调控PI3K/Akt/NF-κB信号通路改善CHD大鼠心肌炎症和氧化应激状况,进而发挥心肌保护作用。

Glycine Attenuates Myocardial Fibrosis in Myocardial Infarction in Rats Partly through Modulating Signal Transducer and Activator of Transcription 3/Nuclear Factor-κB/Transforming Growth Factor-β axis

Objective: Inflammation and fibrosis are strongly associated with each other. Glycine is present in various traditional Chinese medicines and exhibits anti-inflammatory activity. However, the effects of glycine on myocardial fibrosis(MF) in rats with myocardial infarction(MI) have not been reported. The purpose of this study is to investigate the effects of glycine therapy on MF and comprehend its underlying mechanisms. Materials and Methods: Left anterior descending artery ligation-induced MI in Sprague Dawley rats was leveraged to assess the therapeutic effects of Glycine. Rats received either normal saline or glycine(0.5 mg/g bodyweight) for 7 days. Results: Glycine upregulated cardiac ejection fraction and fractional shortening to improve cardiac function, as evaluated by echocardiography. Histological and immunohistochemical analyses demonstrated that glycine could decrease inflammatory cell infiltration and alleviate collagen deposition. Western blotting revealed that nuclear factor-κB(NF-κB)-mediated inflammatory signaling was also downregulated by glycine treatment. The expression of signal transducer and activator of transcription 3(STAT3), tumor necrosis factor-α, and transforming growth factor-β(TGF-β) was decreased significantly in the glycine-treated group compared to the model group. Thus, glycine plays a protective role against myocardial ischemia and subsequent MF. Conclusion: The protective effects of glycine were achieved partly through STAT3/NF-κB/TGF-β signaling pathway.

TLR4/NF-κB/NLRP3通路抑制炎症反应作用的研究

Toll样受体(TLRs)是参与非特异性免疫的一类重要的蛋白质,是表达与固有免疫细胞最重要的模式识别受体之一,可识别来源于微生物的具有保守结构的分子,即病原体相关分子模式(PAMAs),从而激活机体产生免疫应答。核因子κB(NF-κB)在几乎所有类型的细胞和组织中都有表达,在调节对外部刺激的显著多样性的反应中起着关键作用,因此是多个生理和病理过程中的关键因素。而核苷酸结合寡聚化结构域样受体蛋白3(NLRP3),即NLRP3炎性小体通路在脑缺血再灌注和焦亡中具有重要意义,如NLRP3在脑缺血再灌注损伤时在小胶质细胞中被激活,随后在神经元和微血管内皮细胞中表达。由TLRs激活、NF-κB传递、NLRP3启动形成的TLR/NF-κB/NLRP3信号通路在机体各器官炎症反应中起到关键性作用,本文探讨了各类药物通过抑制该通路炎症反应,起到保护身体各大重要器官的作用。

IKBKE、YAP1和TEAD2在结直肠癌中的表达及临床意义

目的探讨核因子κb激酶亚基ε的抑制剂(IKBKE)、Yes相关蛋白1(YAP1)和转录增强结构域转录因子2(TEAD2)在结直肠癌(CRC)组织中的表达及其临床意义。方法收集2016年1月至2017年12月在蚌埠医科大学第一附属医院手术切除的142例CRC组织及对应癌旁组织,采用免疫组化法检测标本中IKBKE、YAP1和TEAD2的表达情况。分析3种蛋白在CRC组织中表达的相关性,分析蛋白阳性率与患者临床病理参数及预后的关系;绘制Kaplan-Meier生存曲线,比较这些蛋白不同表达情况患者的生存差异。采用Cox回归分析影响患者预后的危险因素。结果CRC组织中IKBKE、YAP1和TEAD2的阳性率均显著高于癌旁组织(65.5%比9.9%,73.9%比14.1%,66.9%比8.5%,均P<0.05)。IKBKE的表达与肿瘤的分化程度、浸润深度、淋巴结转移、肿瘤-淋巴结-远处转移(TNM)分期有关,YAP1和TEAD2的表达均与肿瘤的分化程度、浸润深度、淋巴结转移、远处转移及TNM分期有关。Spearman秩相关分析显示CRC组织中IKBKE与YAP1、TEAD2表达均呈正相关(均P<0.01)。Kaplan-Meier生存分析显示IKBKE、YAP1和TEAD2阳性表达组的总生存率降低。Cox回归分析显示IKBKE、YAP1和TEAD2阳性、肿瘤分化程度高、TNM分期高是CRC患者预后的独立危险因素。结论CRC中IKBKE、YAP1和TEAD2阳性表达与肿瘤的分化程度、TNM分期、转移等因素有关,可能成为CRC治疗的潜在靶点;检测这3个蛋白的表达有助于评估预后。

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.

Hepatitis B virus X protein up-regulates tumor necrosis factor-α expression in cultured mesangial cells via ERKs and NF-κB pathways

Objective:To investigate the effects of hepatitis B virus(HBV)X protein(HBx)on the expression of tumor necrosis factor-α(TNF-α)in glomerular mesangial cells(GMCs)and the underlying intracellular signal pathways.Methods:The plasmid pCI-neo-X that carries the X gene of hepatitis B virus was transfected into cultured GMCs.HBx expression in the transfected GMCs was assessed by Western-blot.TNF-αprotein and mRNA were assessed by ELISA and semi-quantitative RT-PCR,respectively.Three kinase inhibitors-U0126,an inhibitor of extracellular signal-regulated kinases(ERKs);lactacvstin,an inhibitor of nuclear factor-κB(NF-κB);and SB203580,a selective inhibitor of p38 MAP kinase(p38 MAPK)were used to determine which intracellular signal pathways may underlie the action of HBx on TNF-αexpression in transfected GMCs.Results:A significant increase in HBx expression in pCI-neo-X transfected GMCs was detected at 36 h and 48 h,which was not affected by any of those kinase inhibitors mentioned above.A similar increase in the expression of both TNF-αprotein and mRNA was also observed at 36 h and 48 h,which was significantly decreased in the presence of U0126 or lactacytin,but not SB203580.Conclusions:HBx upregulates TNF-αexpression in cultured GMCs,possibly through ERKs and NF-κB pathway,but not p38 MAPK pathway.

Breviscapine attenuates acute pancreatitis by inhibiting expression of PKCα and NF-κB in pancreas

AIM:To study the effect of breviscapine (Bre) on activity of protein kinase Cα (PKCα) and nuclear factor (NF)-κB in pancreas,and the mechanism of Bre attenuating acute pancreatitis (AP). METHODS:One hundred and eight rats were randomly divided into acute necrotizing pancreatitis (ANP) group,Bre group (ANP + Bre group) and sham operation (SO) group,36 rats in each group. ANP model was induced by a retrograde injection of 4% sodium deoxycholate into the bilio-pancreatic duct. Fifteen minutes after the ANP model was induced,the rats in Bre group were intraperitoneally injected with Bre (0.4 mg/100 g body weight or 0.1 mL/100 g body weight). Survival time and mortality of rats were calculated. Serum amylase and malondialdehyde levels were measured,volume of ascites was recorded and morphology of pancreas and lung was evaluated at 1,5 and 10 h,after the ANP model was induced,respectively. Expressions of PKCα and subunit p65 of NF-κB in pancreas were detected by immunohistochemistry and Western blotting. RESULTS:The life span of rats was longer and the mortality was lower in Bre group than in ANP group 13.51 ± 5.46 vs 25.36 ± 8.11 (P < 0.05). The amylase and MDA levels as well as the volume of ascites were lower and the pathological changes in pancreas and lung were less in Bre group than ANP group (P < 0.05),indicating that the pancreatitis is less severe in Bre group than ANP group. The activation of PKCα and NF-κB p65 in pancreas was induced rapidly and reached their peak at 1 h or 5 h after ANP,but their activity in Bre group was significantly inhibited. CONCLUSION:Bre exerts its therapeutic effect on AP by inhibiting the activation of PKCα and NF-κB p65 in pancreas.

Clinicopathologic significance of expression of nuclear factor-kB RelA and its target gene products in gastric cancer patients

AIM:To assess the prognostic significance of nuclear factor-kB (NF-kB) and its target genes in gastric cancer. METHODS:The tumor tissues of 115 patients with gastric cancer were immunohistochemically evaluated using monoclonal antibodies against NF-kB RelA. Preoperative serum levels of vascular endothelial growth factor (VEGF), interleukin-6 (IL-6) were assessed via enzyme-linked immuno-sorbent assay. C-reactive protein (CRP) and serum amyloid A (SAA) were measured via immunotrubidimetry. RESULTS:Positive rate of NF-kB RelA was 42.6%. NF-kB RelA expression in tumor tissues was also related to serum levels of IL-6 (P = 0.044) and CRP (P = 0.010). IL-6, SAA, CRP were related to depth of invasion, VEGF and SAA were correlated with lymph node metastasis. IL-6, VEGF, SAA and CRP were related to the stage. Univariate analysis demonstrated that immunostaining of NF-kB RelA, levels of IL-6, VEGF, SAA were significantly related with both disease free survival and over-all survival (OS). Multivariate analysis verified that NF-kB RelA [hazard ratio (HR): 3.40, P = 0.024] and SAA (HR: 3.39, P = 0.045) were independently associated with OS. CONCLUSION: Increased expression of NF-kB RelA and high levels of serum SAA were associated with poor OS in gastric cancer patients.

Crotalaria ferruginea extract attenuates lipopolysaccharide-induced acute lung injury in mice by inhibiting MAPK/NF-κB signaling pathways

Objective:To evaluate the anti-inflammatory activity of Crotalaria ferruginea extract(CFE)and its mechanism.Methods:An intratracheal lipopolysaccharide(LPS)instillationinduced acute lung injury(ALI)model was used to study the antiinflammatory activity of CFE in vivo.The LPS-induced shock model was used to analyze the effect of CFE on survival.LPS-stimulated RAW264.7 cell model was used to investigate the anti-inflammatory activity of CFE in vitro and the effects on mitogen-activated protein kinase(MAPK)or nuclear factor-κB(NF-κB)signaling pathways.Results:CFE administration decreased the number of inflammatory cells,reduced the levels of tumor necrosis factor-α(TNF-α),monocyte chemotactic protein-1(MCP-1),interleukin-6(IL-6),and interferon-γ,and diminished protein content in the bronchoalveolar lavage fluid of mice.CFE also reduced lung wet-to-dry weight ratio,myeloperoxidase,and lung tissue pathological injury.CFE preadministration improved the survival rate of mice challenged with a lethal dose of LPS.CFE reduced LPS-activated RAW264.7 cells to produce nitric oxide,TNF-α,MCP-1,and IL-6.Furthermore,CFE inhibited nuclear translocation and phosphorylation of NF-κB P65,extracellular signal-regulated kinase,c-Jun N-terminal kinases,and P38 MAPKs.Conclusions:CFE exhibits potent anti-inflammatory activity in LPS-induced ALI mice,LPS-shock mice,and RAW264.7 cells,and its mechanism may be associated with the inhibition of NF-κB and MAPK signaling pathways.Crotalaria ferruginea may be a useful therapeutic drug for the treatment of ALI and other respiratory inflammations.

桥本甲状腺炎合并甲状腺乳头状癌患者中Toll样受体3和核转录因子-κB的表达及相关性分析

目的分析桥本甲状腺炎(Hashimoto thyroiditis,HT)合并甲状腺乳头状癌(PTC)患者Toll样受体3(toll-like receptor 3,TLR3)和核转录因子-κB(nuclear transcription factor-κB,NF-κB)表达及相关性。方法收取邯郸市中心医院2020年3月~2022年3月收治的130例行手术切除的甲状腺标本,其中正常甲状腺组织标本43例,HT标本47例,HT合并PTC标本40例,分析TLR3和NF-κB在正常甲状腺组织、HT组、HT合并PTC组中的表达,分析HT合并PTC组中TLR3和NF-κB表达与临床病理参数关系,Pearson相关性分析TLR3和NF-κB的关系。结果TLR3在正常甲状腺组织、HT组、HT合并PTC组中的阳性表达率分别为0(0/43)、80.85%(38/47)、90.00%(36/40);NF-κB在以上三组中的阳性表达率分别为0(0/43)、68.09%(32/47)、85.00%(34/40)。TLR3和NF-κB在HT组、HT合并PTC组中的阳性表达率均高于正常甲状腺组织(P<0.05),TLR3和NF-κB表达与性别、年龄、HT合并PTC病理学特征、病灶类型、淋巴结转移、甲状腺包膜侵犯差异比较均无统计学意义(P均>0.05)。TLR3和NF-κB呈显著正相关(r=0.589,P<0.05)。结论TLR3和NF-κB在HT合并PTC组织中的阳性率高于正常甲状腺组织,且二者表达呈正相关。

HSPA1A受HOTAIR/miR-590-3p轴调控促进I/R诱导的心肌梗死大鼠体内NF-κB介导的炎症反应

目的:探讨热休克70 kDa蛋白1A(HSPA1A)受长链非编码RNA(lncRNA)高温转录物反义RNA(HOTAIR)/微小RNA(miR)-590-3p轴的调控,对心肌缺血/再灌注(I/R)诱导的急性心肌梗死大鼠模型的调控作用。方法:建立大鼠心肌I/R损伤模型,将SD大鼠随机分为6组,分别为假手术组、I/R组、I/R联合siRNA沉默的HSPA1A组(I/R+si-HSPA1A组)、I/R联合siRNA沉默的HOTAIR组(I/R+si-HOTAIR组)、I/R联合si-HSPA1A的阴性对照组(I/R+si-NC组)、I/R联合siRNA沉默的HOTAIR的阴性对照组(I/R+si-NC2组)。建立H9C2细胞缺氧/复氧(H/R)模型,将H9C2细胞随机分为Ctrl组、H/R组、H/R联合si-HOTAIR组(H/R+si-HOTAIR组)、H/R联合si-HOTAIR和过表达HSPA1A组(I/R+si-HOTAIR+pc-HSPA1A组)4组。将常规培养的H9C2细胞分为miR-590-3p的拟似物组(mimic组)、mimic阴性对照组(mimic-NC组)、si-NC2组和si-HOTAIR组4组。用苏木素-伊红(HE)染色进行心肌组织病理学检查。用酶联免疫吸附实验(ELISA)检测心肌组织中肌酸激酶MB(CK-MB)、心肌肌钙蛋白I(cTnI)的表达以及血清中白细胞介素(IL)-6、IL-1β、肿瘤坏死因子-α(TNF-α)的浓度。四甲基偶氮唑蓝(MTT)法检测H9C2细胞的活力。用Western blot分析HSPA1A、核因子-κB(NF-κB)P65、磷酸化的NF-κB P65(p-NF-κB P65)、IL-6、IL-1β、TNF-α的表达。用实时定量PCR(RT-qPCR)法分析HOTAIR和miR-590-3p的表达。利用双荧光素酶报告实验检测miR-590-3p与HSPA1A的3′非翻译区(UTR)以及HOTAIR与miR-590-3p的结合作用。结果:与假手术组比,I/R诱导后明显增加了心肌组织损伤和炎症(均P<0.05)。与I/R+si-NC组比,I/R+si-HSPA1A组的HSPA1A、NF-κB P65、p-NF-κB P65、IL-6、IL-1β、TNF-α表达水平都下调(均P<0.05),缓解了心肌损伤和炎症反应。与假手术组比,I/R诱导后明显增加了HOTAIR表达并下调了miR-590-3p的表达(均P<0.05)。与I/R+si-NC2组比,I/R+si-HOTAIR组的HSPA1A、NF-κB P65、p-NF-κB P65、IL-6、IL-1β、TNF-α表达水平都下调(均P<0.05),miR-590-3p的表达水平上调(P<0.05),缓解了心肌损伤和炎症反应。与H/R组比,H/R+si-HOTAIR组上调了H/R诱导下的H9C2细胞的增殖率(P<0.05),而H/R+si-HOTAIR+pc-HSPA1A组的细胞增殖率明显低于H/R+si-HOTAIR组(P<0.05)。与H/R组比,H/R+si-HOTAIR组H/R诱导的H9C2细胞中HOTAIR、HSPA1A、IL-1β、IL-6和TNF-α的表达显著降低(均P<0.05),而H/R+si-HOTAIR+pc-HSPA1A组IL-1β、IL-6和TNF-α的表达则显著增加(均P<0.05)。双荧光素酶报告实验检测发现HSPA1A的3′-UTR可直接结合miR-590-3p(P<0.05),且在H9C2细胞中,与mimic-NC组比,mimic组中的HSPA1A蛋白表达明显下调(P<0.05)。另外,HOTAIR也可直接结合miR-590-3p(P<0.05)。在H9C2细胞中,与si-NC2组相比,si-HOTAIR组的miR-590-3p表达水平上调(P<0.05)。结论:HSPA1A受HOTAIR/miR-590-3p轴调控促进I/R诱导的心肌梗死大鼠体内NF-κB介导的炎症反应。

Maraviroc promotes recovery from traumatic brain injury in mice by suppression of neuroinflammation and activation of neurotoxic reactive astrocytes

Neuroinflammation and the NACHT,LRR,and PYD domains-containing protein 3 inflammasome play crucial roles in secondary tissue damage following an initial insult in patients with traumatic brain injury(TBI).Maraviroc,a C-C chemokine receptor type 5 antagonist,has been viewed as a new therapeutic strategy for many neuroinflammatory diseases.We studied the effect of maraviroc on TBI-induced neuroinflammation.A moderate-TBI mouse model was subjected to a controlled cortical impact device.Maraviroc or vehicle was injected intraperitoneally 1 hour after TBI and then once per day for 3 consecutive days.Western blot,immunohistochemistry,and TUNEL(terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling)analyses were performed to evaluate the molecular mechanisms of maraviroc at 3 days post-TBI.Our results suggest that maraviroc administration reduced NACHT,LRR,and PYD domains-containing protein 3 inflammasome activation,modulated microglial polarization from M1 to M2,decreased neutrophil and macrophage infiltration,and inhibited the release of inflammatory factors after TBI.Moreover,maraviroc treatment decreased the activation of neurotoxic reactive astrocytes,which,in turn,exacerbated neuronal cell death.Additionally,we confirmed the neuroprotective effect of maraviroc using the modified neurological severity score,rotarod test,Morris water maze test,and lesion volume measurements.In summary,our findings indicate that maraviroc might be a desirable pharmacotherapeutic strategy for TBI,and C-C chemokine receptor type 5 might be a promising pharmacotherapeutic target to improve recovery after TBI.

Therapeutic proteasome inhibition in experimental acute pancreatitis

AIM： To establish the therapeutic potential of proteasome inhibition, we examined the therapeutic effects of MG132 （Z-Leu-Leu-Leu-aldehyde） in an experimental model of acute pancreatitis. METHODS： Pancreatitis was induced in rats by two hourly intraperitoneal （ip） injections of cholecystokinin octapeptide （CCK; 2×100μg/kg） and the proteasome inhibitor MG132 （10 mg/kg ip） was administered 30 min after the second CCK injection. Animals were sacrificed 4 h after the first injection of CCK. RESULTS： Administering the proteasome inhibitor MG132 （at a dose of 10 mg/kg, ip） 90 min after the onset of pancreatic inflammation induced the expression of cell-protective 72 kDa heat shock protein （HSP72） and decreased DNA-binding of nuclear factor-κB （NF-κB）. Furthermore MG132 treatment resulted in milder inflammatory response and cellular damage, as revealed by improved laboratory and histological parameters of pancreatitis and associated oxidative stress. CONCLUSION： Our findings suggest that proteasome inhibition might be beneficial not only for the prevention, but also for the therapy of acute pancreatitis.

TLR4 signaling and the inhibition of liver hepcidin expression by alcohol

AIM: To understand the role of toll-like receptor 4 (TLR4) signaling in the regulation of iron-regulatory hormone, hepcidin by chronic alcohol consumption.

L-and T-type Ca^(2+)channels dichotomously contribute to retinal ganglion cell injury in experimental glaucoma

Retinal ganglion cell apoptotic death is the main pathological characteristic of glaucoma,which is the leading cause of irreversible blindness.Disruption of Ca^(2+)homeostasis plays an important role in glaucoma.Voltage-gated Ca^(2+)channel blockers have been shown to improve vision in patients with glaucoma.However,whether and how voltage-gated Ca^(2+)channels are involved in retinal ganglion cell apoptotic death are largely unknown.In this study,we found that total Ca^(2+)current densities in retinal ganglion cells were reduced in a rat model of chronic ocular hypertension experimental glaucoma,as determined by whole-cell patch-clamp electrophysiological recordings.Further analysis showed that L-type Ca^(2+)currents were downregulated while T-type Ca^(2+)currents were upregulated at the later stage of glaucoma.Western blot assay and immunofluorescence experiments confirmed that expression of the Ca_(V)1.2 subunit of L-type Ca^(2+)channels was reduced and expression of the Ca_(V)3.3 subunit of T-type Ca^(2+)channels was increased in retinas of the chronic ocular hypertension model.Soluble tumor necrosis factor-α,an important inflammatory factor,inhibited the L-type Ca^(2+)current of isolated retinal ganglion cells from control rats and enhanced the T-type Ca^(2+)current.These changes were blocked by the tumor necrosis factor-αinhibitor XPro1595,indicating that both types of Ca^(2+)currents may be mediated by soluble tumor necrosis factor-α.The intracellular mitogen-activated protein kinase/extracellular signal-regulated kinase pathway and nuclear factor kappa-B signaling pathway mediate the effects of tumor necrosis factor-α.TUNEL assays revealed that mibefradil,a T-type calcium channel blocker,reduced the number of apoptotic retinal ganglion cells in the rat model of chronic ocular hypertension.These results suggest that T-type Ca^(2+)channels are involved in disrupted Ca^(2+)homeostasis and apoptosis of retinal ganglion cells in glaucoma,and application of T-type Ca^(2+)channel blockers,especially a specific CaV3.3 blocker,may be a potential strategy for the treatment of glaucoma.