Hemorrhagic transformation in patients with large-artery atherosclerotic stroke is associated with the gut microbiota and lipopolysaccharide

Hemorrhagic transformation is a major complication of large-artery atheroscle rotic stroke(a major ischemic stro ke subtype)that wo rsens outcomes and increases mortality.Disruption of the gut microbiota is an important feature of stroke,and some specific bacteria and bacterial metabolites may contribute to hemorrhagic transformation pathogenesis.We aimed to investigate the relationship between the gut microbiota and hemorrhagic transformation in largearte ry atheroscle rotic stro ke.An observational retrospective study was conducted.From May 2020 to September 2021,blood and fecal samples were obtained upon admission from 32 patients with first-ever acute ischemic stroke and not undergoing intravenous thrombolysis or endovascular thrombectomy,as well as 16 healthy controls.Patients with stro ke who developed hemorrhagic transfo rmation(n=15)were compared to those who did not develop hemorrhagic transformation(n=17)and with healthy controls.The gut microbiota was assessed through 16S ribosomal ribonucleic acid sequencing.We also examined key components of the lipopolysaccharide pathway:lipopolysaccharide,lipopolysaccharide-binding protein,and soluble CD14.We observed that bacterial diversity was decreased in both the hemorrhagic transformation and non-hemorrhagic transfo rmation group compared with the healthy controls.The patients with ischemic stro ke who developed hemorrhagic transfo rmation exhibited altered gut micro biota composition,in particular an increase in the relative abundance and dive rsity of members belonging to the Enterobacteriaceae family.Plasma lipopolysaccharide and lipopolysaccharide-binding protein levels were higher in the hemorrhagic transformation group compared with the non-hemorrhagic transfo rmation group.lipopolysaccharide,lipopolysaccharide-binding protein,and soluble CD14 concentrations were associated with increased abundance of Enterobacte riaceae.Next,the role of the gut microbiota in hemorrhagic transformation was evaluated using an experimental stroke rat model.In this model,transplantation of the gut microbiota from hemorrhagic transformation rats into the recipient rats triggered higher plasma levels of lipopolysaccharide,lipopolysaccharide-binding protein,and soluble CD14.Ta ken togethe r,our findings demonstrate a noticeable change in the gut microbiota and lipopolysaccharide-related inflammatory response in stroke patients with hemorrhagic transformation.This suggests that maintaining a balanced gut microbiota may be an important factor in preventing hemorrhagic transfo rmation after stro ke.

Quercetin Alleviates Lipopolysaccharide-Induced Cardiac Inflammation via Inhibiting Autophagy and Programmed Cell Death

Objective The aim of this study is to explore the potential modulatory role of quercetin against Endotoxin or lipopolysaccharide(LPS)induced septic cardiac dysfunction.Methods Specific pathogen-free chicken embryos(n=120)were allocated untreated control,phosphate buffer solution(PBS)vehicle,PBS with ethanol vehicle,LPS(500 ng/egg),LPS with quercetin treatment(10,20,or 40 nmol/egg,respectively),Quercetin groups(10,20,or 40 nmol/egg).Fifteenday-old embryonated eggs were inoculated with abovementioned solutions via the allantoic cavity.At embryonic day 19,the hearts of the embryos were collected for histopathological examination,RNA extraction,real-time polymerase chain reaction,immunohistochemical investigations,and Western blotting.Results They demonstrated that the heart presented inflammatory responses after LPS induction.The LPS-induced higher mRNA expressions of inflammation-related factors(TLR4,TNFα,MYD88,NF-κB1,IFNγ,IL-1β,IL-8,IL-6,IL-10,p38,MMP3,and MMP9)were blocked by quercetin with three dosages.Quercetin significantly decreased immunopositivity to TLR4 and MMP9 in the treatment group when compared with the LPS group.Quercetin significantly decreased protein expressions of TLR4,IFNγ,MMP3,and MMP9 when compared with the LPS group.Quercetin treatment prevented LPS-induced increase in the mRNA expression of Claudin 1 and ZO-1,and significantly decreased protein expression of claudin 1 when compared with the LPS group.Quercetin significantly downregulated autophagyrelated gene expressions(PPARα,SGLT1,APOA4,AMPKα1,AMPKα2,ATG5,ATG7,Beclin-1,and LC3B)and programmed cell death(Fas,Bcl-2,CASP1,CASP12,CASP3,and RIPK1)after LPS induction.Quercetin significantly decreased immunopositivity to APOA4,AMPKα2,and LC3-II/LC3-I in the treatment group when compared with the LPS group.Quercetin significantly decreased protein expressions of AMPKα1,LC3-I,and LC3-II.Quercetin significantly decreased the protein expression to CASP1 and CASP3 by immunohistochemical investigation or Western blotting in treatment group when compared with LPS group.Conclusion Quercetin alleviates cardiac inflammation induced by LPS through modulating autophagy,programmed cell death,and myocardiocytes permeability.

Foeniculum vulgare Mill.inhibits lipopolysaccharide-induced microglia activation and ameliorates neuroinflammation-mediated behavioral deficits in mice

Objective:To investigate the effect of Foeniculum vulgare extract against lipopolysaccharide(LPS)-induced microglial activation in vitro as well as cognitive behavioral deficits in mice.Methods:LPS-activated BV-2 cell viability was measured using MTT assay and reactive oxygen species(ROS)was studied using DCF-DA assay.The antioxidative enzymes and pro-inflammatory mediators were analyzed using respective ELISA kits and Western blotting.For in vivo testing,LPS(1 mg/kg,i.p.)was given daily for five days in male Swiss albino mice to produce chronic neuroinflammation.Cognitive and behavioral tests were performed using open-field,passive avoidance,and rotarod experiments in LPS-induced mice.Results:Foeniculum vulgare extract(25,50 and 100μg/mL)significantly attenuated the LPS-activated increase in nitric oxide(NO),ROS,cyclooxygenase-2,inducible NO synthase,IL-6,and TNF-alpha(P<0.05).Moreover,LPS-induced oxidative stress and reduced antioxidative enzyme levels were significantly improved by Foeniculum vulgare extract(P<0.05).The extract also regulated the NF-κB/MAPK signaling in BV-2 cells.In an in vivo study,Foeniculum vulgare extract(50,100,and 200 mg/kg)markedly mitigated the LPS-induced cognitive and locomotor impairments in mice.The fingerprinting analysis showed distinctive peaks with rutin,kaempferol-3-O-glucoside,and anethole as identifiable compounds.Conclusions:Foeniculum vulgare extract can ameliorate LPS-stimulated neuroinflammatory responses in BV-2 microglial cells and improve cognitive and locomotor performance in LPS-administered mice.

Antidepressant effects of Peiyuan Jieyu formula in a mouse model of chronic stress in conjunction with lipopolysaccharide-induced depression

Objective:To explore the mechanism of the Peiyuan Jieyu formula in treating depression by assessing its impact on a lipopolysaccharide-induced(LPS-induced)depression mouse model.Methods:We created a mouse model of depression by exposing mice that had previously received chronic stress to intraperitoneal LPS injections.The mice were divided into the following groups:control,model,fluoxetine,Tiansi Yin,Sini powder,and low-,medium-,and high-dose Peiyuan Jieyu formula groups.Forced swim and tail suspension tests were used to assess the efficacy of the depression(despair)model,and weight gain rates were also measured.Furthermore,serum levels of various depression and inflammation-associated molecules,including tumor necrosis factor-a(TNF-a),interferon-γ(IFN-γ),tryptophan,5-hydroxytryptamine,kynurenine(KYN),and kynurenic acid(KA)were assessed.Furthermore,the expression levels of ionic calcium-binding adaptor molecule-1(IBA-1)and indoleamine 2,3-dioxygenase(IDO)mRNA in hippocampal microglia were measured.Results:The model group displayed greater despair-associated immobility,which was shortened in response to various doses of Peiyuan Jieyu formula.Furthermore,formula administration significantly reduced serum TNF-a levels and hippocampal IDO mRNA expression.The high formula dose also reduced IFN-γand IBA-1 levels,the latter was also decreased in response to the medium formula dose.However,the low formula dose reduced serum KYN level and KYN/tryptophan(TRP)and KYN/KA ratios.Conclusion:The Peiyuan Jieyu formula holds immense potential in treating depression in a mouse model,potentially inhibiting inflammation and improving TRP-KYN metabolic disorders.

Sinapic acid ameliorates D-galactosamine/lipopolysaccharideinduced fulminant hepatitis in rats:Role of nuclear factor erythroidrelated factor 2/heme oxygenase-1 pathways

BACKGROUND Sinapic acid(SA)has been shown to have various pharmacological properties such as antioxidant,antifibrotic,anti-inflammatory,and anticancer activities.Its mechanism of action is dependent upon its ability to curb free radical production and protect against oxidative stress-induced tissue injuries.AIM To study the hepatoprotective effects of SA against lipopolysaccharide(LPS)/Dgalactosamine(D-GalN)-induced acute liver failure(ALF)in rats.METHODS Experimental ALF was induced with an intraperitoneal(i.p.)administration of 8μg LPS and 800 mg/kg D-GalN in normal saline.SA was administered orally once daily starting 7 d before LPS/D-GalN treatment.RESULTS Data showed that SA ameliorates acute liver dysfunction,decreases serum levels of alanine transaminase(ALT),and aspartate aminotransferase(AST),as well as malondialdehyde(MDA)and NO levels in ALF model rats.However,pretreatment with SA(20 mg/kg and 40 mg/kg)reduced nuclear factor kappalight-chain-enhancer of activated B cells(NF-κB)activation and levels of inflammatory cytokines(tumor necrosis factor-αand interleukin 6).Also,SA increased the activity of the nuclear factor erythroid-related factor 2/heme oxygenase-1(Nrf2/HO-1)signaling pathway.CONCLUSION In conclusion,SA offers significant protection against LPS/D-GalN-induced ALF in rats by upregulating Nrf2/HO-1 and downregulating NF-κB.

Adiponectin deficiency exacerbates lipopolysaccharide/ D-galactosamine-induced liver injury in mice

瞄准：在 Kupffer 房间的功能上检验 adiponectin 的效果，脂肪的多糖(LPS ) 的关键调节的人导致了肝损伤。方法：D-galactosamine (GalN ) 和 LPS intraperitoneally 被注入 adiponectin-/- 老鼠和野类型老鼠。Kupffer 房间，从 Sprague-Dawley 孤立老鼠，是 preincubated 与或没有 adiponectin，然后与 LPS 对待。结果：在猛烈老鼠， GalN/LPS 注射显著地降低了幸存率，显著地提起了丙氨酸转氨酶和肿瘤坏死 factor-alpha (TNF-alpha ) 的血浆层次并且显著地减少了与野类型老鼠相比的 IL-10 层次。在肝的 TNF-alpha 基因表示是哪个更高并且 IL-10 的那些比在野类型老鼠在猛烈老鼠是更低的。在有教养的 adiponectin-pre-treated Kupffer 房间， LPS 显著地降低了 TNF-alpha 层次并且在培养基和他们的各自的基因表示层次提起了 IL-10 层次，与没有 adiponectin-pre-treatment 的 Kupffer 房间相比。结论：Adiponectin supresses TNF-alpha 生产并且响应 LPS 刺激由 Kupffer 房间导致 IL-10 生产，并且 adiponectin 的缺乏提高导致 LPS 的肝损伤。

Inhibiting the expression of hepatocyte nuclear factor 4 alpha attenuates lipopolysaccharide/ D-galactosamine-induced fulminant hepatic failure in mice

BACKGROUND: Hepatocyte nuclear factor 4 alpha (HNF4α) plays an important role in regulating cytokine-induced inflammatory responses. This study aimed to investigate the role of HNF4α in the development of fulminant hepatic failure (FHF) induced by lipopolysaccharide/D-galactosamine (LPS/D-GalN). METHODS: The FHF model was induced by simultaneous intraperitoneal injection of LPS/D-GalN in mice. Three days prior to LPS/D-GalN administration, HNF4α short-hairpin interfering RNA expression plasmid or physiological saline was injected via the tail vein with a hydrodynamics-based procedure. The degree of hepatic damage and cumulative survival rate were subsequently assessed. RESULTS: The expression of HNF4α was increased in the early stage after LPS/D-GalN administration. Inhibiting the expression of HNF4α reduced serum levels of alanine aminotransferase and aspartate aminotransferase, alleviated histological injury, and improved the survival of mice with FHF. In addition, both serum and hepatic tumor necrosis factor alpha expression were suppressed when HNF4α expression was inhibited in mice with FHF. CONCLUSION: Inhibiting HNF4α expression protects mice from FHF induced by LPS/D-GalN, but the exact mechanism behind this needs further investigation.

Protective Activity of <i>Markhamia tomentosa</i>(Benth.) K. Schum. (Bignoniaceae) Methanol Leaves Extract against D-Galactosamine/Lipopolysaccharide-Induced Acute Liver Injury in Mice

Markhamia tomentosa (Benth.) K. Schum. (Mt) is a Cameroonian medicinal plant, traditionally used to treat painful and inflammatory illness. This study aimed to examine the effects of methanol leaves extract (MLE) of Mt in D-galactosamine (D-GaIN)/lipopolysaccharide (LPS)-induced liver injury. The MLE (100 and 200 mg/kg), Ascorbic acid (10 mg/kg) and distilled water were administered 12 h and 1 h before intraperitoneal injection of D-GaIN (10 mg/mouse)/LPS (0.1 μg/g). Animals were sacrificed 6 h after D-GalN/LPS challenge. Liver injury was assessed biochemically by determination of aspartate aminotransferase (ASAT), alanine aminotransferase (ALAT), alkaline phosphatase (ALP), superoxide dismutase (SOD) and catalase activities. Malondialdehyde (MDA), reduced glutathione (GSH), nitrites, total protein and bilirubin levels were explored. Histopathological examination of liver tissue was also performed. Liver enzymes (ALAT, ASAT, ALP) activity, nitrites, MDA and bilirubin levels were increased, while protein level, SOD and catalase activities were significantly reduced by D-GalN/LPS administration. MLE (100 or 200 mg/kg) protected mice against D-GalN/LPS-induced death. In addition, the plant extract significantly reduced ALAT and ALP activity, exhibiting 23.00% and 62.20% protection, respectively. SOD activity and total protein were significantly (p < 0.05) increased by the plant extract. Total bilirubin and MDA levels were reduced (p < 0.01) by 37.75% and 62.79%, respectively in animal treated with MLE. Histological analysis of liver sections showed that MLE (100 or 200 mg/kg) protected mice against D-GaIN/LPS-induced liver injury. The obtained results showed that MLE of Mt may possess hepatoprotective effects. Protection afforded by MLE against D-GalN/LPS-induced fulminant liver injury may result from reduction of oxidative stress.

Enhanced expressions and activations of leukotriene C4synthesis enzymes in D-galactosamine/lipopolysaccharide-induced rat fulminant hepatic failure model

AIM: To investigate the expression and activity of leukotriene C4 (LTC4) synthesis enzymes and their underlying relationship with cysteinyl leukotriene (cys-LT) generation in a rat fulminant hepatic failure (FHF) model induced by D-galactosamine/lipopolysaccharide (D-GaIN/ LPS). METHODS: Rats were treated with D-GaIN (300 mg/kg) plus LPS (0.1 mg/kg) for 1, 3, 6, and 12 h. Enzyme immunoassay was used to determine the hepatic cys-LT content. Reverse transcription-polymerase chain reaction (RT-PCR), Western blot or immunohistochemical assay were employed to assess the expression or location of LTC4 synthesis enzymes, which belong to membrane associated proteins in eicosanoid and glutathione (MAPEG) metabolism superfamily. Activity of LTC4 synthesis enzymes was evaluated by determination of the products of LTA4 after incubation with liver microsomes using high performance liquid chromatography (HPLC). RESULTS: Livers were injured after treatment with D-GalN/LPS, accompanied by cys-LT accumulation at the prophase of liver injury. Both LTC4 synthase (LTC4S) and microsomal glutathione-S-transferase (mGST) 2 were expressed in the rat liver, while the latter was specifically located in hepatocytes. Their mRNA and protein expressions were up-regulated at an earlier phase after treatment with D-GalN/LPS. Meantime, a higher activity of LTC4 synthesis enzymes was detected, although the activity of LTC4S played the main role in this case. CONCLUSION: The expression and activity of both LTC4S and mGST2 are up regulated in a rat FHF model, which are, at least, partly responsible for cys-LT hepatic accumulation.

Yeast hydrolysate attenuates lipopolysaccharide-induced inflammatory responses and intestinal barrier damage in weaned piglets

Background Intestinal inflammation is the main risk factor causing intestinal barrier dysfunction and lipopolysaccharide(LPS)can trigger inflammatory responses in various eukaryotic species.Yeast hydrolysate(YH)possesses multibiological effects and is received remarkable attention as a functional ingredient for improving growth performance and promoting health in animals.However,there is still inconclusive on the protective effects of dietary YH supplementation on intestinal barrier of piglets.This study was conducted to investigate the attenuate effects of YH supplementation on inflammatory responses and intestinal barrier injury in piglets challenged with LPS.Methods Twenty-four piglets(with an average body weight of 7.42±0.34 kg)weaned at 21 days of age were randomly assigned to one of two dietary treatments(12 replications with one pig per pen):a basal diet or a basal diet containing YH(5 g/kg).On the 22nd d,6 piglets in each treatment were intraperitoneally injected with LPS at 150μg/kg BW,and the others were injected with the same amount of sterile normal saline.Four hours later,blood samples of each piglet were collected and then piglets were euthanized.Results Dietary YH supplementation increased average daily feed intake and average daily gain(P<0.01),decreased the ratio of feed intake to gain of piglets(P sponse,evidenced by the increase o=0.048).Lipopolysaccharide(LPS)injection induced systemic inflammatory ref serum concentrations of haptoglobin(HP),adrenocorticotropic hormone(ACTH),cortisol,and interleukin-1β(IL-1β).Furthermore,LPS challenge resulted in inflammatory intestinal damage,by up-regulation of the protein or mRNA abundances of tumor necrosis factor-α(TNF-α),IL-1β,toll-like receptors 4(TLR4)and phosphor-nuclear factor-κB-p65(p-NFκB-p65)(P<0.01),and down-regulation of the jejunal villus height,the protein and mRNA abundances of zonula occludens-1(ZO-1)and occludin(OCC;P<0.05)in jejunal mucosa.Dietary YH supplementation decreased the impaired effects of ACTH,cortisol,HP,IL-1βand diamine oxidase in serum(P<0.05).Moreover,YH supplementation also up-regulated the jejunal villus height,protein and mRNA abundances of ZO-1 and OCC(P<0.05),down-regulated the mRNA expressions of TNF-αand IL-1βand the protein abundances of TNF-α,IL-1β,TLR4 and p-NFκB-p65 in jejunal mucosa in LPS-challenged pigs(P<0.01).Conclusion Yeast hydrolysate could attenuate inflammatory response and intestinal barrier injury in weaned piglets challenged with LPS,which was associated with the inhibition of TLR4/NF-κB signaling pathway activation.

Effect of dual targeting procyanidins nanoparticles on metabolomics of lipopolysaccharide-stimulated inflammatory macrophages

Inflammation plays an important role in the occurrence and development of many inflammatory diseases.The purpose of this study was to evaluate the anti-inflammatory effect and metabolic behavior of the dual targeting procyanidins(PC)nanoparticles on lipopolysaccharide(LPS)-stimulated inflammatory macrophages by metabolomics method.The double-targeting PC nanoparticles could specifi cally target both the CD44 receptor and mitochondria,while the single targeting PC-loaded nanoparticles that could target the CD44 receptor on the surface of macrophages.The double-targeting PC nanoparticles had better inhibitory effect than single-targeting PC nanoparticles on the leakage of lactate dehydrogenase and reactive oxygen species overexpression induced by LPS.Amino acid metabolism,energy metabolism and purine metabolism were disordered in LPS-treated group,and metabolic pathway analysis indicated that the double-targeting PC nanoparticles reversed some of LPS impacts.The changes of these potential biomarkers and their corresponding pathways are helpful to further understand the mechanism of PC nanoparticles in alleviating inflammation,and promote their application in nutrition intervention.

Lipopolysaccharide-induced Trigeminal Ganglion Nerve Fiber Damage is Associated with Autophagy Inhibition

Objective This study aimed to determine whether lipopolysaccharide(LPS)induces the loss of corneal nerve fibers in cultured trigeminal ganglion(TG)cells,and the underlying mechanism of LPS-induced TG neurite damage.Methods TG neurons were isolated from C57BL/6 mice,and the cell viability and purity were maintained for up to 7 days.Then,they were treated with LPS(1µg/mL)or the autophagy regulator(autophibib and rapamycin)alone or in combination for 48 h,and the length of neurites in TG cells was examined by the immunofluorescence staining of the neuron-specific proteinβ3-tubulin.Afterwards,the molecular mechanisms by which LPS induces TG neuron damage were explored.Results The immunofluorescence staining revealed that the average length of neurites in TG cells significantly decreased after LPS treatment.Importantly,LPS induced the impairment of autophagic flux in TG cells,which was evidenced by the increase in the accumulation of LC3 and p62 proteins.The pharmacological inhibition of autophagy by autophinib dramatically reduced the length of TG neurites.However,the rapamycin-induced activation of autophagy significantly lessened the effect of LPS on the degeneration of TG neurites.Conclusion LPS-induced autophagy inhibition contributes to the loss of TG neurites.

Structural modifications of Helicobacter pylori lipopolysaccharide:An idea for how to live in peace

In this review,we discuss the findings and concepts underlying the"persistence mechanisms"of Helicobacter pylori(H.pylori),a spiral-shaped,Gram-negative rod bacterium that was discovered as a gastric pathogen by Marshall and Warren in 1984.H.pylori colonizes the gastric mucosa of nearly half of the human population.Infections appear in early childhood and,if not treated,persist for life.The presence or absence of symptoms and their severity depend on multiple bacterial components,host susceptibility and environmental factors,which allow H.pylori to switch between pathogenicity and commensalism.Many studies have shown that H.pylori components may facilitate the colonization process and the immune response of the host during the course of H.pylori infection.These H.pylori-driven interactions might result from positive or negative modulation.Among the negative immunomodulators,a prominent position is occupied by a vacuolating toxin A(VacA)and cytotoxin-associated gene A(CagA)protein.However,in light of the recent studies that are presented in this review,it is necessary to enrich this panel with H.pylori lipopolysaccharide(LPS).Together with CagA and VacA,LPS suppresses the elimination of H.pylori bacteria from the gastric mucosa by interfering with the activity of innate and adaptive immune cells,diminishing the inflammatory response,and affecting the adaptive T lymphocyte response,thus facilitating the development of chronic infections.The complex strategy of H.pylori bacteria for survival in the gastric mucosa of the host involves both structural modifications of LPS lipid A to diminish its endotoxic properties and the expression and variation of Lewis determinants,arranged in O-specific chains of H.pylori LPS.By mimicking host components,this phenomenon leaves these bacteria"invisible"to immune cells.Together,these mechanisms allow H.pylori to survive and live for many years within their hosts.

Raising on Water Stocking Density Reduces Geese Reproductive Performances via Water Bacteria and Lipopolysaccharide Contaminations in “Geese-Fish” Production System

This study was carried out to unravel the mechanism of reductions in production performances in high stocking density geese flocks during summer months in ＂geese-fish＂ production system. Experiment 1 observed the water bacterial growth, lipopolysaccharde concentrations in water and geese blood, and geese reproductive performances from summer to winter, in two flocks with varying on water stocking densities. Results showed that counts of total bacteria, Escherichia coli and Salmonella in water, as well as water and geese plasma LPS concentrations, exhibited a tendency decreasing from the highest levels in summer, to intermediate levels in autumn, and to the lowest values in winter. Such seasonal decreases in bacteria and LPS concentrations were associated with similar seasonal decreases in embryo mortality during incubation. In addition, embryos dead or showing development retardation by day 25 of incubation contained copious LPS in allantoic fluid, in contrast to the negligible amount in normal developing embryos. Raising on water stocking density elevated bacteria counts, LPS concentrations in water and geese plasma, and decreased egg fertility but increased embryo mortality during incubation. In experiment 2, exogenous LPS treatment to the geese depressed egg laying, reduced egg hatchability, caused sickness behavior in the goslings hatched. In experiment 3, exogenous LPS directly administered to day 8 and 18 embryos during incubation dose dependently increased mortality and decreased hatchability, and caused sickness behavior in the goslings hatched. It is concluded that the raising on water geese stocking density stimulates pathogenic bacteria growth in water, which via LPS contamination impaires embryo development in incubation and therefore reduces geese reproductive performance and gosling quality during the hot summer months.

Sanguinarine Attenuates Lipopolysaccharide-induced Inflammation and Apoptosis by Inhibiting the TLR4/NF-KB Pathway in H9c2 Cardiomyocytes

The inflammatory response is involved in the pathogenesis of the most common types of heart disease. Sanguinarine （SAN） has various pharmacological properties such as anti-inflammatory, antioxidant, antibacterial, antitumor, and immune-enhancing properties. However, few studies have investigated the effects of SAN on lipopolysaceharide （LPS）-induced inflammatory and apoptotic responses in H9c2 cardiomyocytes. Therefore, in this study, H9c2 cells were co-treated with SAN and LPS, and the mRNA levels of pro-inflammation markers and the apoptosis rate were measured to clarify the effect of SAN on cardiac inflammation. The underlying mechanism was further investigated by detecting the activation of Toll-like receptor （TLR）4/nuclear faetor-κB （NF-κB） signaling pathways. As a result, increased mRNA expression of interleukin （IL）-1β, IL-6, and TNFα induced by LPS was attenuated after SAN treatment; LPS-induced apoptosis ofHge2 cardiomyocytes and cleaved-caspase 8, 9, 3 were all significantly reduced by SAN. Further experiments showed that the beneficial effect of SAN on blocking the inflammation and apoptosis of H9c2 cardiomyocytes induced by LPS was associated with suppression of the TLR4/NF-κB signaling pathway. It was suggested that SAN suppressed the LPS-induced inflammation and apoptosis of H9c2 cardiomyocytes, which may be mediated by inhibition of the TLR4/NF-κB signaling pathway. Thus, SAN may be a feasible therapy to treat sepsis patients with cardiac dysfunction.

The anti-inflammatory effects of asiatic acid in lipopolysaccharide-stimulated human corneal epithelial cells

AIM： To investigate the anti-inflammatory effects of asiatic acid（AA） on lipopolysaccharide（LPS）-induced inflammatory response in human corneal epithelial cells（HCECs）.METHODS： Cell viability was measured using a cell counting kit-8（CCK-8） assay.Quantitative real-time polymerase chain reaction（qR T-PCR） was used to determine the mR NA expression of interleukin-8（IL-8）,interleukin-6（IL-6）,interleukin-1β（IL-1β）,tumor necrosis factor-alpha（TNF-α）,and transforming growth factor-β（TGF-β） in HCECs.Intracellular reactive oxygen species（ROS） was measured using the ROS assay kit.Glutathione（GSH） concentration was measured using the total GSH assay kit.Akt1 and Akt phosphorylation（p-Akt1） levels were measured by Western blotting and immunofluorescence.RESULTS： AA induced toxicity at high concentrations and significantly stimulated the proliferation of HCECs at concentrations of 20 μmol/L for 1h.LPS at concentrations of 300 ng/mL for 1h significantly stimulated the mR NA expression of IL-8,IL-6,IL-1β,TNF-α,and TGF-β in HCECs,while the stimulation effects were significantly inhibited by AA（20 μmol/L）.In addition,AA was found to decrease the content of ROS,increase GSH generation,and also inhibit LPS-induced p-Akt in HCECs.CONCLUSION： AA decreases the generation of inflammatory factors IL-8,IL-6,IL-1β,TNF-α,and TGF-β in LPSstimulated HCECs.AA significantly inhibites the intracellular concentrations of ROS and increases GSH generation.AA also inhibites LPS-induced p-Akt in HCECs.These findings reveal that AA has anti-inflammation effects in LPS-stimulated HCECs.

Ethanol extracts of Scutellaria baicalensis protect against lipopolysaccharide-induced acute liver injury in mice

Objective: To investigated the protective potential of ethanol extracts of Scutellaria baicalensis(S. baicalensis) against lipopolysaccharide(LPS)-induced liver injury. Methods: Dried roots of S. baicalensis were extracted with ethanol and concentrated to yield a dry residue. Mice were administered 200 mg/kg of the ethanol extracts orally once daily for one week. Animals were subsequently administered a single dose of LPS(5 mg/kg of body weight, intraperitoneal injection). Both protein and m RNA levels of cytokines, such as tumor necrosis factor alpha, interleukin-1β, and interleukin-6 in liver tissues were evaluated by ELISA assay and quantitative PCR. C yclooxygenase-2, inducible nitric oxide synthase, and nuclear factor-κB protein levels in liver tissues were analyzed by western blotting. Results: Liver injury induced by LPS signifi cantly increased necrosis factor alpha, interleukin-1β, interleukin-6, cyclooxygenase-2, inducible nitric oxide synthase, and nuclear factor-κB in liver tissues. Treatment with ethanol extracts of S. baicalensis prevented all of these observed changes associated with LPS-induced injury in liver mice.Conclusions: Our study showed that S. baicalensis is potentially protective against LPS-induced liver injury in mice.

Hyperoside Suppresses Lipopolysaccharide-induced Inflammation and Apoptosis in Human Umbilical Vein Endothelial Cells

Finding the novel drug from the effective components of traditional Chinese herbal medicine is a hotspot of the modem pharmacological research. Hyperoside （HYP） belongs to flavonoid glycosides, and it has various properties, such as anti-inflammation, anti-spasm, anti-diuretic, antitussive, lowering blood pressure, and lowering cholesterol effects as well as protective effects for the cardiac and cerebral blood vessels. The purpose of this study was to investigate the effects of HYP on inflammatory and apoptotic responses in vascular endothelial cells stimulated by lipopolysaccharide （LPS） and further to identify the possible mechanisms underlying these effects. In our study, human umbilical vein endothelial cells （HUVECs） were stimulated with 1 Bg/mL LPS in the presence or absence of HYP （10, 20 and 50 μmol/L）. Our results indicated that HYP alone exerted no cytotoxicity on HUVECs, while it had an upregulatory effect on the viability of HUVECs induced by LPS in a dose-dependent manner; increased mRNA expression of IL-1β, IL-6, TNFα and iNOS induced by LPS was attenuated after treatment with HYP both in a dose- and time-dependent manner; LPS-induced HUVECs apoptosis and cleaved-caspase 8, 9, 3 were all significantly reduced by HYP. Furthermore, the possible pathway involved in apoptosis and inflammation by HYP was detected, and the results showed that when treated with HYP, LPS-induced mitochondrial membrane instability was significantly inhibited through up-regulation of Bcl-2 and down-regulation of Bax. Furthermore, the expression of TLR4 and the phosphorylation of IκBα and p65 in LPS-treated cells were blocked by HYP. Our results suggested that HYP treatment prevented HUVECs from LPS- induced inflammation and apoptosis responses, which might be mediated by inhibiting TLR4/ NFκB pathway.

Effects of Meglumine Cyclic Adenylate Pretreatment on Systemic inflammatory Response Syndrome Induced by Lipopolysaccharide in Rats

Studies showed that the use of cyclic adenosine monophosphate(cAMP) substitutes or intracellular c AMP activators increased intracellular cAMP level, causing anti-inflammatory effects. This study was to investigate the effects of pretreatment with meglumine cyclic adenylate(MCA), a compound of meglumine and cAMP, on systemic inflammation induced by lipopolysaccharide(LPS) in rats. Eighteen adult male Sprague-Dawley rats were randomly divided into 3 groups(n=6 each): control group(NS group), LPS group(LPS group) and LPS with MCA pretreatment group(MCA group). Systemic inflammation was induced with LPS 10 mg/kg injected via the femoral vein in LPS and MCA groups. In MCA group, MCA 2 mg/kg was injected via the femoral vein 20 min before LPS injection, and the equal volume of normal saline was given in NS and LPS groups at the same time. Three hours after LPS injection, the blood samples were taken from the abdominal aorta for determination of plasma concentrations of TNF-α, IL-1, IL-6, IL-10, cAMP by ELISA and NF-κBp65 expression by Western blotting. The experimental results showed that inflammatory and antiinflammatory indices were increased in LPS group compared to NS group; inflammatory indices were declined and anti-inflammatory indices were increased in MCA group relative to LPS group. Our study suggested that MCA pretreatment may attenuate LPS-induced systemic inflammation.

Lipopolysaccharide inhibits or accelerates biomedical titanium corrosion depending on environmental acidity

Titanium and its alloys are routinely used as biomedical implants and are usually considered to be corrosion resistant under physiological conditions. However, during inflammation, chemical modifications of the peri-implant environment including acidification occur. In addition certain biomolecules including lipopolysaccharide（LPS）, a component of Gram-negative bacterial cell walls and driver of inflammation have been shown to interact strongly with Ti and modify its corrosion resistance. Gram-negative microbes are abundant in biofilms which form on dental implants. The objective was to investigate the influence of LPS on the corrosion properties of relevant biomedical Ti substrates as a function of environmental acidity. Inductively coupled plasma mass spectrometry was used to quantify Ti dissolution following immersion testing in physiological saline for three common biomedical grades of Ti（ASTM Grade 2, Grade 4 and Grade 5）. Complementary electrochemical tests including anodic and cathodic polarisation experiments and potentiostatic measurements were also conducted. All three Ti alloys were observed to behave similarly and ion release was sensitive to p H of the immersion solution. However, LPS significantly inhibited Ti release under the most acidic conditions（p H 2）, which may develop in localized corrosion sites, but promoted dissolution at p H 4–7, which would be more commonly encountered physiologically.The observed pattern of sensitivity to environmental acidity of the effect of LPS on Ti corrosion has not previously been reported. LPS is found extensively on the surfaces of skin and mucosal penetrating Ti implants and the findings are therefore relevant when considering the chemical stability of Ti implant surfaces in vivo.