AIM: To study the inhibition effect of TAK-242 on the proliferation of rat eye Tenon's capsule fibroblasts via the toll-like receptor 4(TLR4) signaling pathway.METHODS: SD rat Tenon's capsule fibroblasts were ...AIM: To study the inhibition effect of TAK-242 on the proliferation of rat eye Tenon's capsule fibroblasts via the toll-like receptor 4(TLR4) signaling pathway.METHODS: SD rat Tenon's capsule fibroblasts were extracted and cultured, then the cells were divided into normal control group, lipopolysaccharide(LPS) group(10 g/m L LPS) and TAK-242 group(1 μmol/L TAK-242, and 10 μg/m L LPS after 30 min). The expressions of TLR4, transforming growth factor-β1(TGF-β1) and interleukin-6(IL-6) in each group were detected by Western blot and reverse transcriptase-polymerase chain reaction(RT-PCR). Cell proliferation was detected by cell counting kit-8(CCK-8).RESULTS: Double immunofluorescent labeling in the extracted cells showed negative keratin staining and positive vimentin staining. Western blot showed that the LPS group had the highest expression of TLR4 and TGF-β1(P<0.01). Enzyme linked immunosorbent assay(ELISA) also showed that the secretion of IL-6 was the highest in LPS group(P<0.01). But there was no significant difference in TLR4 and TGF-1, as well as IL-6 expressions between the TAK-242 group and the normal control group(P>0.05). RT-PCR showed that the IL-6 m RNA expression in LPS group was the highest in the three groups(P<0.01). CONCLUSION: TAK-242 inhibits the proliferation of LPSinduced Tenon's capsule fibroblasts and the release of inflammatory factors by regulating the TLR4 signalingpathway, providing a new idea for reducing the scarring of the filter passage after glaucoma filtration surgery.展开更多
目的探讨Toll样受体⁃4(Toll like receptor⁃4,TLR⁃4)抑制剂TAK⁃242对大鼠重度牙周炎骨质吸收的影响,为重度牙周炎寻找辅助治疗手段提供实验基础。方法18只3周龄雄性Wistar大鼠随机分为3组(n=6),其中1组为正常对照组,另外2组以含有牙龈...目的探讨Toll样受体⁃4(Toll like receptor⁃4,TLR⁃4)抑制剂TAK⁃242对大鼠重度牙周炎骨质吸收的影响,为重度牙周炎寻找辅助治疗手段提供实验基础。方法18只3周龄雄性Wistar大鼠随机分为3组(n=6),其中1组为正常对照组,另外2组以含有牙龈卟啉单胞菌(P.gingivalis)ATCC33277的5⁃0丝线结扎大鼠双侧上颌磨牙行重度牙周炎建模,分为牙周炎组、TAK⁃242组;TAK⁃242组从丝线结扎第1天起,通过尾静脉隔天注射1次溶于DMSO的TAK⁃242(2 mg/kg),另外两组注射相同体质量比例的DMSO溶剂,连续8周;第8周末处死3组大鼠,获取大鼠上颌骨标本,采用micro⁃CT扫描后三维重建,测量特定位点釉牙骨质界⁃牙槽嵴顶的距离评估骨丧失量,并对牙槽骨骨质相关参数和骨质微结构进行分析;组织学切片苏木精⁃伊(HE)染色观察牙周组织病理改变;甲基绿染色观察牙槽骨吸收情况;抗酒石酸酸性磷酸酶(TRAP)染色观察破骨细胞分布情况。结果Micro⁃CT定量分析显示:牙周炎组与TAK⁃242组牙槽骨吸收显著高于对照组;与牙周炎组相比,TAK⁃242组大鼠上颌第一磨牙近、远中根吸收位点的骨丧失均显著减轻(P<0.001),骨密度(P<0.05)与骨体积/总体积分数(P<0.01)显著增高,骨小梁数目与骨小梁厚度(P<0.01)相对增多,骨小梁分离度(P<0.01)和骨小梁结构模式指数显著降低。牙周炎组骨质呈现疏松多孔的蜂窝状结构,骨小梁结构恶化,向杆状结构转变;TAK⁃242组骨质微结构改善,骨量改善,骨小梁分布相对更致密,骨小梁结构与对照组更相似。HE染色发现牙周炎组与TAK⁃242组牙周附着丧失与牙槽骨吸收较对照组显著;与牙周炎组相比,甲基绿染色表明TAK⁃242组骨吸收减轻,TRAP染色显示破骨细胞浸润减少(P<0.001)。结论TLR⁃4抑制剂TAK⁃242能缓解大鼠重度牙周炎骨吸收,改善其多孔、稀疏、排列紊乱的炎症性骨小梁结构。展开更多
目的探讨Toll样受体4(TLR4)抑制剂TAK-242对糖尿病周围神经痛(DPN)模型大鼠的治疗效果及其可能的作用机制。方法 SPF级雄性SD大鼠60只,随机均分为3组,分别为正常对照组(NC组)、DPN模型组(DPN组)、TAK-242治疗组(TAK组)。采用链脲佐菌素(...目的探讨Toll样受体4(TLR4)抑制剂TAK-242对糖尿病周围神经痛(DPN)模型大鼠的治疗效果及其可能的作用机制。方法 SPF级雄性SD大鼠60只,随机均分为3组,分别为正常对照组(NC组)、DPN模型组(DPN组)、TAK-242治疗组(TAK组)。采用链脲佐菌素(STZ)方法建立DPN大鼠模型,采用ELISA及RT-PCR方法检测DPN模型大鼠腰膨大脊髓组织的高迁移率族蛋白B1(HMGB1)-TLR4轴上下游基因(HMGB1、TLR4、MAPK、NF-κB、IL-6)的变化,分析上述细胞因子表达水平与大鼠疼痛行为的相关性。使用TLR4抑制剂TAK-242对DPN模型大鼠进行药物干预,观察其治疗效果及对HMGB1-TLR4轴基因表达的影响。结果 ELISA检测显示,DPN组大鼠的血清HMGB1、TLR4、IL-6表达水平均较NC组升高(P<0.05);TAK组大鼠的TLR4及IL-6表达水平较DPN组下降(P<0.05)。RT-PCR检测显示,DPN组大鼠的HMGB1、TLR4、NF-κB、IL-6 m RNA表达水平较NC组升高(P<0.05);TAK组大鼠的TLR4、NF-κB、IL-6 m RNA表达水平较DPN组下降(P<0.05)。结论 TLR4抑制剂TAK-242通过阻断HMGB1-TLR4轴对DPN模型动物起治疗作用。展开更多
Secondary brain damage caused by hyperactivation of autophagy and inflammatory responses in neurons plays an important role in hypoxic-ischemic brain damage(HIBD).Although previous studies have implicated Toll-like re...Secondary brain damage caused by hyperactivation of autophagy and inflammatory responses in neurons plays an important role in hypoxic-ischemic brain damage(HIBD).Although previous studies have implicated Toll-like receptor 4(TLR4)and nuclear factor kappa-B(NF-κB)in the neuroinflammatory response elicited by brain injury,the role and mechanisms of the TLR4-mediated autophagy signaling pathway in neonatal HIBD are still unclear.We hypothesized that this pathway can regulate brain damage by modulating neuron autophagy and neuroinflammation in neonatal rats with HIBD.Hence,we established a neonatal HIBD rat model using the Rice-Vannucci method,and injected 0.75,1.5,or 3 mg/kg of the TLR4 inhibitor resatorvid(TAK-242)30 minutes after hypoxic ischemia.Our results indicate that administering TAK-242 to neonatal rats after HIBD could significantly reduce the infarct volume and the extent of cerebral edema,alleviate neuronal damage and neurobehavioral impairment,and decrease the expression levels of TLR4,phospho-NF-κB p65,Beclin-1,microtubule-associated protein l light chain 3,tumor necrosis factor-α,and interleukin-1βin the hippocampus.Thus,TAK-242 appears to exert a neuroprotective effect after HIBD by inhibiting activation of autophagy and the release of inflammatory cytokines via inhibition of the TLR4/NF-κB signaling pathway.This study was approved by the Laboratory Animal Ethics Committee of Affiliated Hospital of Yangzhou University,China(approval No.20180114-15)on January 14,2018.展开更多
基金Supported by National Natural Science Foundation Program of China (No.81770920)Hubei Health and Family Planning Commission Youth Talent Project (No. WJ2017Q037)
文摘AIM: To study the inhibition effect of TAK-242 on the proliferation of rat eye Tenon's capsule fibroblasts via the toll-like receptor 4(TLR4) signaling pathway.METHODS: SD rat Tenon's capsule fibroblasts were extracted and cultured, then the cells were divided into normal control group, lipopolysaccharide(LPS) group(10 g/m L LPS) and TAK-242 group(1 μmol/L TAK-242, and 10 μg/m L LPS after 30 min). The expressions of TLR4, transforming growth factor-β1(TGF-β1) and interleukin-6(IL-6) in each group were detected by Western blot and reverse transcriptase-polymerase chain reaction(RT-PCR). Cell proliferation was detected by cell counting kit-8(CCK-8).RESULTS: Double immunofluorescent labeling in the extracted cells showed negative keratin staining and positive vimentin staining. Western blot showed that the LPS group had the highest expression of TLR4 and TGF-β1(P<0.01). Enzyme linked immunosorbent assay(ELISA) also showed that the secretion of IL-6 was the highest in LPS group(P<0.01). But there was no significant difference in TLR4 and TGF-1, as well as IL-6 expressions between the TAK-242 group and the normal control group(P>0.05). RT-PCR showed that the IL-6 m RNA expression in LPS group was the highest in the three groups(P<0.01). CONCLUSION: TAK-242 inhibits the proliferation of LPSinduced Tenon's capsule fibroblasts and the release of inflammatory factors by regulating the TLR4 signalingpathway, providing a new idea for reducing the scarring of the filter passage after glaucoma filtration surgery.
文摘目的探讨Toll样受体⁃4(Toll like receptor⁃4,TLR⁃4)抑制剂TAK⁃242对大鼠重度牙周炎骨质吸收的影响,为重度牙周炎寻找辅助治疗手段提供实验基础。方法18只3周龄雄性Wistar大鼠随机分为3组(n=6),其中1组为正常对照组,另外2组以含有牙龈卟啉单胞菌(P.gingivalis)ATCC33277的5⁃0丝线结扎大鼠双侧上颌磨牙行重度牙周炎建模,分为牙周炎组、TAK⁃242组;TAK⁃242组从丝线结扎第1天起,通过尾静脉隔天注射1次溶于DMSO的TAK⁃242(2 mg/kg),另外两组注射相同体质量比例的DMSO溶剂,连续8周;第8周末处死3组大鼠,获取大鼠上颌骨标本,采用micro⁃CT扫描后三维重建,测量特定位点釉牙骨质界⁃牙槽嵴顶的距离评估骨丧失量,并对牙槽骨骨质相关参数和骨质微结构进行分析;组织学切片苏木精⁃伊(HE)染色观察牙周组织病理改变;甲基绿染色观察牙槽骨吸收情况;抗酒石酸酸性磷酸酶(TRAP)染色观察破骨细胞分布情况。结果Micro⁃CT定量分析显示:牙周炎组与TAK⁃242组牙槽骨吸收显著高于对照组;与牙周炎组相比,TAK⁃242组大鼠上颌第一磨牙近、远中根吸收位点的骨丧失均显著减轻(P<0.001),骨密度(P<0.05)与骨体积/总体积分数(P<0.01)显著增高,骨小梁数目与骨小梁厚度(P<0.01)相对增多,骨小梁分离度(P<0.01)和骨小梁结构模式指数显著降低。牙周炎组骨质呈现疏松多孔的蜂窝状结构,骨小梁结构恶化,向杆状结构转变;TAK⁃242组骨质微结构改善,骨量改善,骨小梁分布相对更致密,骨小梁结构与对照组更相似。HE染色发现牙周炎组与TAK⁃242组牙周附着丧失与牙槽骨吸收较对照组显著;与牙周炎组相比,甲基绿染色表明TAK⁃242组骨吸收减轻,TRAP染色显示破骨细胞浸润减少(P<0.001)。结论TLR⁃4抑制剂TAK⁃242能缓解大鼠重度牙周炎骨吸收,改善其多孔、稀疏、排列紊乱的炎症性骨小梁结构。
文摘目的探讨Toll样受体4(TLR4)抑制剂TAK-242对糖尿病周围神经痛(DPN)模型大鼠的治疗效果及其可能的作用机制。方法 SPF级雄性SD大鼠60只,随机均分为3组,分别为正常对照组(NC组)、DPN模型组(DPN组)、TAK-242治疗组(TAK组)。采用链脲佐菌素(STZ)方法建立DPN大鼠模型,采用ELISA及RT-PCR方法检测DPN模型大鼠腰膨大脊髓组织的高迁移率族蛋白B1(HMGB1)-TLR4轴上下游基因(HMGB1、TLR4、MAPK、NF-κB、IL-6)的变化,分析上述细胞因子表达水平与大鼠疼痛行为的相关性。使用TLR4抑制剂TAK-242对DPN模型大鼠进行药物干预,观察其治疗效果及对HMGB1-TLR4轴基因表达的影响。结果 ELISA检测显示,DPN组大鼠的血清HMGB1、TLR4、IL-6表达水平均较NC组升高(P<0.05);TAK组大鼠的TLR4及IL-6表达水平较DPN组下降(P<0.05)。RT-PCR检测显示,DPN组大鼠的HMGB1、TLR4、NF-κB、IL-6 m RNA表达水平较NC组升高(P<0.05);TAK组大鼠的TLR4、NF-κB、IL-6 m RNA表达水平较DPN组下降(P<0.05)。结论 TLR4抑制剂TAK-242通过阻断HMGB1-TLR4轴对DPN模型动物起治疗作用。
基金financially supported by the National Natural Science Foundation of China,No.81771625(to XF)the Jiangsu Provincial Key Medical Discipline of China,No.ZDXKA2016013(to XF)the Pediatric Clinical Center of Suzhou City of China,No.Szzx201504(to XF)
文摘Secondary brain damage caused by hyperactivation of autophagy and inflammatory responses in neurons plays an important role in hypoxic-ischemic brain damage(HIBD).Although previous studies have implicated Toll-like receptor 4(TLR4)and nuclear factor kappa-B(NF-κB)in the neuroinflammatory response elicited by brain injury,the role and mechanisms of the TLR4-mediated autophagy signaling pathway in neonatal HIBD are still unclear.We hypothesized that this pathway can regulate brain damage by modulating neuron autophagy and neuroinflammation in neonatal rats with HIBD.Hence,we established a neonatal HIBD rat model using the Rice-Vannucci method,and injected 0.75,1.5,or 3 mg/kg of the TLR4 inhibitor resatorvid(TAK-242)30 minutes after hypoxic ischemia.Our results indicate that administering TAK-242 to neonatal rats after HIBD could significantly reduce the infarct volume and the extent of cerebral edema,alleviate neuronal damage and neurobehavioral impairment,and decrease the expression levels of TLR4,phospho-NF-κB p65,Beclin-1,microtubule-associated protein l light chain 3,tumor necrosis factor-α,and interleukin-1βin the hippocampus.Thus,TAK-242 appears to exert a neuroprotective effect after HIBD by inhibiting activation of autophagy and the release of inflammatory cytokines via inhibition of the TLR4/NF-κB signaling pathway.This study was approved by the Laboratory Animal Ethics Committee of Affiliated Hospital of Yangzhou University,China(approval No.20180114-15)on January 14,2018.
