Long-term oral Asperosaponin VI attenuates cardiac dysfunction,myocardial fibrosis in a rat model of chronic myocardial infarction

The aim of the study was to determine the effects of Asperosaponin VI (ASA VI), a triterpene saponin isolated from Dipsacus asper Wall, on chronic myocardial infarction (MI) and possible mechanisms in rats. MI was induced by permanent ligation of the left coronary artery. Twenty-four hours after MI, the rats were administered the extract by gavage (once a day). Six weeks after MI/sham surgery, cardiac dysfunction, infarct size (IS), cardiac fibrosis, hydroxyproline concentration, the oxidative stress parameter and inflammation mediators were examined. The results indicated that ASA VI improved left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), ±dP/dt, heart weight/body weight, right ventricular weight/body weight and lung weight/body weight (P < 0.01, P < 0.05). These were accompanied by the attenuation of cardiac fibrosis, IS and hydroxyproline concentration (P < 0.01, P < 0.05). ASA VI could decrease the levels of tumor necrosis factor-α (TNF-α) and interleukin 6 (IL-6), but increase IL-10 content (P < 0.01, P < 0.05). Furthermore, it also could raise the activities of catalase, superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), but reduce malonyldialdehyde (MDA) level (P < 0.01, P < 0.05). The results indicated that ASA VI improved cardiac function and myocardial fibrosis from myocardial ischemia injury, and this cardioprotection might be attributed to reduce oxidative stress and regulate inflammation mediators.     

Inhibition of inflammatory mediators by polyphenolic plant extracts in human intestinal Caco-2 cells

The mitogen-activated protein kinases (MAPK) and nuclear factor κB (NF-κB) are involved in transduction cascades that play a key role in inflammatory response. We tested the ability of preselected natural polyphenolic extracts (grape seed, cocoa, sugar cane, oak, mangosteen and pomegranate) to modulate intestinal inflammation using human intestinal Caco-2 cells treated for 4 h with these extracts and then stimulated by cytokines for 24 or 48 h. The effect of polyphenolic extracts, at 50 μmol of gallic acid equivalent/l, was investigated on inflammation-related cellular events: (i) NF-κB activity (cells transfected with a NF-κB-luciferase construct), (ii) activation of Erk1/2 and JNK (western blotting), (iii) secretion of interleukin 8 (IL-8) (ELISA), (iv) secretion of prostaglandin (PG) E₂ (ELISA), (v) production of NO (Griess method). Results show that: (i) sugar cane, oak and pomegranate extracts inhibited NF-κB activity (from 1.6 to 1.9-fold) (P < 0.001); (ii) pomegranate slightly inhibited Erk1/2 activation (1.3-fold) (P = 0.008); (iii) oak and pomegranate decreased NO synthesis by 1.5-fold (P < 0.001) and that of IL-8 by 10.3 and 6.7-fold respectively; (iv) pomegranate and cocoa decreased PGE₂ synthesis by 4.6 (P < 0.0001) and 2.2-fold (P = 0.001), respectively. We suggest that pomegranate extract could be particularly promising in dietary prevention of intestinal inflammation.     

Inhibiting NF-κB activation and ROS production are involved in the mechanism of silibinin's protection against D-galactose-induced senescence

Aging is featured by intelligence decline, behavioral disorders and cognitive disability. Autophagy is related to senescent development. In this study, we investigated the roles of NF-κB and autophagy in hippocampal neurons of d-galactose-induced senescent mice, and examined the protective roles of silibinin. Senescence was induced in 6-month-old mice by subcutaneous injection of d-galactose (150 mg/kg/d, for 6 weeks). Silibinin (50 mg/kg/d, intramuscular injection, for 6 weeks) or inhibitors (PDTC, 3-MA or rapamycin, 50 mg/kg/d, subcutaneous injection, for 6 weeks) were given 1 h before d-galactose exposure. Senescent control animals received vehicle for the same time. Ethological analysis, immunofluorescence staining, flow cytometric analysis, western blot and enzyme activity assays were used. Compared with senescent controls, silibinin, PDTC or rapamycin-treated mice showed upregulations of spatial recognition memory (P < 0.05), cellular oxidoreductase activities (P < 0.05) and autophagy (P < 0.05) as well as downregulations of MDA (P < 0.05) and ROS (P < 0.05) levels. We propose in d-galactose-induced murine senescence, autophagy is inhibited by NF-κB, inducing the deactivations of celluar oxidoreductases and upregulation of ROS level. The protection by autophagy and the promotion of cellular oxidoreductase activities via inhibiting NF-κB activation and ROS production are involved in the mechanism of silibinin's protection against d-galactose-induced senescence.     

Hydrogen sulfide attenuates lipopolysaccharide-induced cognitive impairment: A pro-inflammatory pathway in rats

The present study investigated the effect of sodium hydrosulfide (NaHS), a H₂S donor, on cognitive impairment and neuroinflammatory changes induced by bilateral intracerebroventricular injections of LPS at a dose of 10 μg/rat. Rats received 5 mg/kg NaHS or volume-matched vehicle administration by intraperitoneal injection 3 days before LPS injection then for 9 days once daily. Morris water maze was used to detect the cognitive function. Compared to the sham-treated rats, LPS injection significantly prolonged the mean escape latency in the navigation test (P < 0.05) and shortened the adjusted escape latency by approximately 30% (P < 0.05). Meanwhile, LPS injection decreased H₂S level but increased pro-inflammatory mediators (i.e., TNF-α, TNFR1, degradation of IκB-α and thereafter activation of NF-κB) in hippocampus. However, these effects of LPS were significantly ameliorated with NaHS treatment (P < 0.05 vs vehicle-treated group). The present data suggest that H₂S attenuates LPS-induced cognitive impairment through reducing the overproduction of pro-inflammatory mediators via inhibition of NF-κB pathways in rats. This study sets the stage for exploring a novel H₂S releasing agent for preventing or retarding the development or progression of neurological disorders such as Alzheimer's disease.     

MG132 treatment attenuates cardiac remodeling and dysfunction following aortic banding in rats via the NF-κB/TGFβ1 pathway

Although MG132, a proteasome inhibitor, is suggested to impede secondary cardiac remodeling after hypertension, the mechanism and optimal duration of treatment remain unknown. This study was designed to investigate the effects and possible mechanism of MG132 on hypertension-induced cardiac remodeling. Male Sprague–Dawley rats subjected to abdominal aortic constriction (AAC) or sham operation received an intraperitoneal injection of MG132 (0.1 mg kg⁻¹ day⁻¹) or vehicle over a 2- or 8-week period. In the end, left ventricular (LV) function was evaluated with echocardiography and pressure tracing. Collagen deposition within the LV myocardium was assessed with Masson's trichrome staining. Ubiquitin-proteasome system (UPS), NF-κB, I-κB, TGFβ1 and Smad2 within the LV tissue were evaluated. In addition, angiotensin II within both plasma and LV tissue was also examined. Compared with the sham groups, the vehicle-treated AAC group exhibited a higher angiotensin II level, LV/body weight ratio, septal and posterior wall thicknesses, and a markedly reduced cardiac function (P < 0.05). Treatment with MG132 for 8 weeks attenuated these cardiac remodeling parameters and improved cardiac function (P < 0.01). 2- and 8-week hypertension led to activation of UPS, which was followed by activation of NF-κB and increased expression of TGFβ1 and Smad2 (P < 0.01). MG132 significantly inhibited NF-κB activity and down-regulate the levels of TGFβ1 and Smad2 expression by 2 and still at 8 weeks (P < 0.01). Short- and long-term treatment with MG132 significantly attenuated hypertension-induced cardiac remodeling and dysfunction, which may be mediated by the NF-κB/TGFβ1 signaling pathway.     

甘草酸对慢性肾脏病大鼠心肌HMGB1/TLR4/NF-κB/HIF-1α信号通路的影响

目的 探讨应用甘草酸（Gly）治疗时对慢性肾脏病（CKD）大鼠心室肌高迁移率族蛋白B1/Toll样受体4/核因子κB/缺氧诱导因子1α(HMGB1/TLR4/NF-κB/HIF-1α)信号通路的影响。方法 将38只Wistar大鼠按随机数字表法分为4组：假手术（Sham）组、Sham+Gly组、CKD组、CKD+Gly组，5/6肾切除制备CKD模型，Gly腹腔注射给药（80 mg/kg）。4周后行血流动力学及心脏彩超观察各组大鼠心脏功能；心室取血检测生化指标；心肌组织取材检测HMGB1、TLR4、NF-κB、HIF-1α蛋白表达的变化。结果 与Sham组相比，CKD组肌酐、尿酸、尿素氮和血清镁水平增高（P<0.05），血压升高，舒张期室间隔厚度、收缩期左心室内径增加，左心室射血分数降低，E/A比值降低，肺动脉血流加速时间延长（P<0.05）;HE染色可见心肌细胞肥大，Masson染色见心肌纤维化程度增加（P<0.05），心肌组织HMGB1、NF-κB、HIF-1α蛋白表达水平上调（P<0.05）。与CKD组相比，CKD+Gly组收缩期室间隔厚度增加，左心室射血分...     

吡咯烷二硫代氨基甲酸盐下调基质金属蛋白酶9表达机制的研究

目的探讨TNF-α诱导的肺泡巨噬细胞(AM)性基质金属蛋白酶9(MMP-9)表达的信号通路以及吡咯烷二硫代氨基甲酸盐(PDTC)对MMP-9表达的影响机制。方法从慢性阻塞性肺疾病患者支气管肺泡灌洗液中分离与培养AM,以PDTC预处理AM,以TNF-α或IL-1刺激AM。半定量逆转录-聚合酶链反应法检测MMP-9 mRNA的表达;Western blot检测MMP-9蛋白的表达及TNF-α或IL-1诱导的IκBα磷酸化水平。凝胶阻滞分析实验检测NF-κB活性。结果TNF-α上调AM源性MMP-9 mRNA和蛋白的表达(P<0.05);PDTC抑制TNF-α诱导的MMP-9的表达(P<0.05)。PDTC对TNF-α或IL-1诱导的IκBα的磷酸化均无抑制作用(P>0.05)。PDTC对TNF-α或IL-1诱导的NF-κB的活化均有抑制作用(P<0.05);且PDTC不能在体外直接抑制NF-κB的DNA结合活性(P>0.05)。结论NF-κB在TNF-α诱导的AM源性MMP-9的表达中起着重要作用;PDTC可能通过抑制泛素化-蛋白酶小体途径来下调TNF-α诱导的AM源性MMP-9的表达。     

Cytokine expression in trichloroethylene-induced hypersensitivity dermatitis: An in vivo and in vitro study

The purpose of this study was to address the association between cytokine expression and the hypersensitivity dermatitis induced by trichloroethylene (TCE) exposure. 28 TCE-induced hypersensitivity dermatitis patients, 22 TCE exposed workers and 22 non-exposed controls were enrolled in the study. The serum levels of interleukin (IL)-1β, IL-6, IL-8 and tumor necrosis factor (TNF)-α were analyzed using a magnetic colorbead-based multiplex assay. The patients showed significantly higher levels of serum IL-1β (p = 0.033 and p = 0.015), IL-6 (p < 0.001), IL-8 (p < 0.001 and p = 0.002) and TNF-α (p = 0.009 and p = 0.005) than the TCE exposed workers and non-exposed controls. There was a significantly positive correlation among these cytokine concentrations, but no significant correlation was found between these cytokine concentrations and the disease duration in patient group. We further compared the effects of trichloroethanol (TCOH) and trichloroacetic acid (TCA), two major metabolites of TCE, on cytokine expression in keratinocyte cell line (HaCaT). IL-1α, IL-6, IL-8 and TNF-α concentrations were tested using enzyme-linked immunosorbent assay (ELISA) after HaCaT cells were treated with different concentrations of TCOH or TCA for 24 h. We found that TCOH, but not TCA, increased the levels of IL-1α and IL-6 in a dose-dependent manner. We also found that TCOH activated the nuclear factor kappa B (NF-κB) pathway. Bay 11-7082 (NF-κB inhibitor) significantly attenuated the TCOH-induced production of IL-6 in HaCaT cells, but IL-1α production was not affected. In conclusions, it is suggested that IL-1β, IL-6, IL-8 and TNF-α were associated with TCE-induced hypersensitivity dermatitis. TCOH induced IL-6 expression through activation of the NF-κB pathway in HaCaT cells and may play an integral role in TCE-induced skin hypersensitivity.     

Synergistic growth inhibition of mouse skin tumors by pomegranate fruit extract and diallyl sulfide: Evidence for inhibition of activated MAPKs/NF-κB and reduced cell proliferation

Limited outcomes from earlier chemopreventive studies have necessitated that some modifications be made to get better efficacy. It is proposed that cancer prevention is more feasible than treatment, and this could be achieved effortlessly with use of multiple agents competent of targeting multiple targets. This study was initiated to examine the chemopreventive efficacy of pomegranate fruit extract (PFE) and diallyl sulfide (DAS), alone and in combination, using 2-stage mouse skin tumorigenesis model. PFE and DAS alone delayed onset and tumor incidence by ∼55% and ∼45%, respectively, while their combination at low doses synergistically decreased tumor incidence more potentially (∼84%, p < 0.01). In addition, regression in tumor volume was seen with continuous combinatorial treatment (p < 0.01). Mechanistic studies revealed that this inhibition was associated with decreased expression of phosphorylated ERK1/2, JNK1 and activated NF-κB/p65, IKKα, IκBα phosphorylation and degradation in skin tissue/tumor. Histological and cell death analysis also confirmed that combined PFE and DAS inhibit cellular proliferation and markedly induce apoptosis than the single agents. Altogether, our results suggest that PFE and DAS in combination impart better suppressive activity than either of these agents alone and provide support that development of novel combination therapies/chemoprevention using dietary agents will be more beneficial against cancer.     

Preventive effect of Coptis chinensis and berberine on intestinal injury in rats challenged with lipopolysaccharides

Coptis chinensis has been used in traditional Chinese medicine to treat inflammatory symptoms. Berberine is the main alkaloid compound of C. chinensis. This study utilized a typical lipopolysaccharide (LPS) injured model to investigate the effects of C. chinensis aqueous extract (CCAE) and berberine (major active ingredient in CCAE) in the gut-derived sepsis. In rats, pretreatment with different doses of berberine (30 or 120 mg/kg bw, i.g.; BBR30 or BBR120) or CCAE (containing 9.9% berberine; 300 mg/kg bw, i.g.; CCAE300) prior to the administration of LPS (20 mg/kg bw, i.p.) significantly suppressed the increased tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and nitrite oxide (NO) in plasma as well as the activation of toll-like receptor 4 (TLR4) and nuclear factor-κB (NF-κB) in ileum. In addition, CCAE300 and BBR30 markedly elevated the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px); significantly prevented the increased malondialdehyde (MDA), NO and villi injury in ileum compared with the negative control. Collectively, CCAE300 and BBR30 reduced the LPS-induced intestinal damage by elevating the activities of SOD and GSH-Px and by suppressing the activation of TLR4 and NF-κB in ileum. These results indicate that CCAE and berberine are promising agents for preventing sepsis and its complications.     

Effect of baicalin on toll-like receptor 4-mediated ischemia/reperfusion inflammatory responses in alcoholic fatty liver condition

Alcoholic fatty liver is susceptible to secondary stresses such as ischemia/reperfusion (I/R). Baicalin is an active component extracted from Scutellaria baicalensis, which is widely used in herbal preparations for treatment of hepatic diseases and inflammatory disorders. This study evaluated the potential beneficial effect of baicalin on I/R injury in alcoholic fatty liver. Rats were fed an alcohol liquid diet or a control isocaloric diet for 5 weeks, and then subjected to 60 min of hepatic ischemia and 5 h of reperfusion. Baicalin (200 mg/kg) was intraperitoneally administered 24 and 1 h before ischemia. After reperfusion, baicalin attenuated the increases in serum alanine aminotransferase activity, tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) levels in alcoholic fatty liver. The increased levels of TNF-α and IL-6 mRNA expression and inducible nitric oxide synthase and cyclooxygenase-2 protein and mRNA expressions increased after reperfusion, which were higher in ethanol-fed animals, were attenuated by baicalin. In ethanol-fed animals, baicalin attenuated the increases in toll-like receptor 4 (TLR4) and myeloid differentiation factor 88 protein expressions and the nuclear translocation of NF-κB after reperfusion. In conclusion, our findings suggest that baicalin ameliorates I/R-induced hepatocellular damage by suppressing TLR4-mediated inflammatory responses in alcoholic fatty liver.     

齐墩果酸调节HMGB1/TLR4/NF-κB介导的炎症通路减轻蛛网膜下腔出血后的早期脑损伤

目的探讨齐墩果酸对大鼠蛛网膜下腔出血模型HMGB1/TLR4/NF-κB炎症通路的影响。方法将72只成年雄性SD大鼠随机分成3组(假手术组、SAH模型组、齐墩果酸20 mg/kg给药组)。采用颈内动脉穿刺法建立大鼠SAH模型,造模1 h后给予齐墩果酸,剂量为20 mg/kg。大鼠SAH模型建立24 h后进行神经功能学评分;检测脑水含量及脑伊文思蓝浸出率;并利用Western blot方法检测脑组织中HMGB1、TLR4、IκBα和p65蛋白的表达量。结果齐墩果酸(20 mg/kg)能够显著增加神经功能评分并且明显降低脑含水量和伊文思蓝渗透率(P<0.01)。齐墩果酸给药可有效降低HMGB1的表达和释放(P<0.01),抑制TLR4的表达、IκBα降解及p65入核(P<0.01)。结论齐墩果酸能够抑制HMGB1/TLR4/NF-κB炎症通路,减轻蛛网膜下腔出血后的炎症反应,改善脑损伤。     

Protective effect of tetrahydrocoptisine against ethanol-induced gastric ulcer in mice

Excessive alcohol consumption can lead to gastric ulcer and the present work was aimed to examine the protective effect of tetrahydrocoptisine (THC) in the model of ethanol-induced gastric ulcer in mice. Fasted mice treated with ethanol 75% (0.5 ml/100 g) were pre-treated with THC (10 or 20 mg/kg, ip), cimetidine (100 mg/kg, ip) or saline in different experimental sets for a period of 3 days, and animals were euthanized 4 h after ethanol ingestion. Gross and microscopic lesions, immunological and biochemical parameters were taken into consideration. The results showed that ethanol induced gastric damage, improving nitric oxide (NO) level, increased pro-inflammatory cytokine (TNF-α and IL-6) levels and myeloperoxidase (MPO) activity, as well as the expression of nuclear factor-κB (NF-κB) in the ethanol group. Pretreatment of THC at doses of 10 and 20 mg/kg bodyweight significantly attenuated the gastric lesions as compared to the ethanol group. These results suggest that the gastroprotective activity of THC is attributed to reducing NO production and adjusting the pro-inflammatory cytokine, inhibited neutrophil accumulation and NF-κB expression.     

阿托伐他汀对兔动脉粥样硬化中NF-κB/IκB和炎症因子表达的影响

目的探讨高脂饮食诱发兔动脉粥样硬化中核因子-κB(NF-κB)的活化与其抑制因子IκB的表达,以及阿托伐他汀对NF-κB/IκB信号途径、ICAM-1和P选择素的影响。方法24只新西兰♂家兔随机分为正常对照组、高胆固醇组和阿托伐他汀组,应用W estern b lot方法检测动脉中胞核NF-κB p65亚基和胞浆IκBα表达变化;免疫放射法检测血清P选择素水平,免疫组织化学检测血管壁ICAM-1的表达。结果高胆固醇组与对照组比较胞核NF-κB p65和血管壁ICAM-1表达明显增强、血P选择素明显升高(P<0.05),胞浆中IκBα表达明显减弱(P<0.05);阿托伐他汀组与高胆固醇组比较NF-κB p65和血管壁ICAM-1明显表达较弱、血P选择素明显减少(P<0.05),胞浆IκBα的表达增强(P<0.05)。结论NF-κB/IκB信号途径在动脉粥样硬化中起重要作用,阿托伐他汀可以减少P选择素、ICAM-1的表达。     

Multiplexed quantitative high content screening reveals that cigarette smoke condensate induces changes in cell structure and function through alterations in cell signaling pathways in human bronchial cells

Human bronchial cells are one of the first cell types exposed to environmental toxins. Toxins often activate nuclear factor-κB (NF-κB) and protein kinase C (PKC). We evaluated the hypothesis that cigarette smoke condensate (CSC), the particulate fraction of cigarette smoke, activates PKC-α and NF-κB, and concomitantly disrupts the F-actin cytoskeleton, induces apoptosis and alters cell function in BEAS-2B human bronchial epithelial cells. Compared to controls, exposure of BEAS-2B cells to doses of 30 μg/ml CSC significantly activated PKC-α, while CSC doses above 20 μg/ml CSC significantly activated NF-κB. As NF-κB was activated, cell number decreased. CSC treatment of BEAS-2B cells induced a decrease in cell size and an increase in cell surface extensions including filopodia and lamellipodia. CSC treatment of BEAS-2B cells induced F-actin rearrangement such that stress fibers were no longer prominent at the cell periphery and throughout the cells, but relocalized to perinuclear regions. Concurrently, CSC induced an increase in the focal adhesion protein vinculin at the cell periphery. CSC doses above 30 μg/ml induced a significant increase in apoptosis in BEAS-2B cells evidenced by an increase in activated caspase 3, an increase in mitochondrial mass and a decrease in mitochondrial membrane potential. As caspase 3 increased, cell number decreased. CSC doses above 30 μg/ml also induced significant concurrent changes in cell function including decreased cell spreading and motility. CSC initiates a signaling cascade in human bronchial epithelial cells involving PKC-α, NF-κB and caspase 3, and consequently decreases cell spreading and motility. These CSC-induced alterations in cell structure likely prevent cells from performing their normal function thereby contributing to smoke-induced diseases.     

Barrier protective effects of withaferin A in HMGB1-induced inflammatory responses in both cellular and animal models

Withaferin A (WFA), an active compound from Withania somnifera, is widely researched for its anti-inflammatory, cardioactive and central nervous system effects. In this study, we first investigated the possible barrier protective effects of WFA against pro-inflammatory responses in human umbilical vein endothelial cells (HUVECs) and in mice induced by high mobility group box 1 protein (HMGB1) and the associated signaling pathways. The barrier protective activities of WFA were determined by measuring permeability, leukocytes adhesion and migration, and activation of pro-inflammatory proteins in HMGB1-activated HUVECs. We found that WFA inhibited lipopolysaccharide (LPS)-induced HMGB1 release and HMGB1-mediated barrier disruption, expression of cell adhesion molecules (CAMs) and adhesion/transendothelial migration of leukocytes to human endothelial cells. WFA also suppressed acetic acid-induced hyperpermeability and carboxymethylcellulose-induced leukocytes migration in vivo. Further studies revealed that WFA suppressed the production of interleukin 6, tumor necrosis factor-α (TNF-α) and activation of nuclear factor-κB (NF-κB) by HMGB1. Collectively, these results suggest that WFA protects vascular barrier integrity by inhibiting hyperpermeability, expression of CAMs, adhesion and migration of leukocytes, thereby endorsing its usefulness as a therapy for vascular inflammatory diseases.