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
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(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
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。
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(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。
