目的研究右美托咪啶(DEX)对内毒素血症大鼠急性肺损伤(ALI)的保护作用,探讨TLR9信号通路相关因子的表达差异。方法 SPF级雄性SD大鼠30只,随机分为3组:假手术组(S组)、内毒素血症组(LPS组)、右美托咪啶组(DEX组),每组10只。LPS组尾静脉...目的研究右美托咪啶(DEX)对内毒素血症大鼠急性肺损伤(ALI)的保护作用,探讨TLR9信号通路相关因子的表达差异。方法 SPF级雄性SD大鼠30只,随机分为3组:假手术组(S组)、内毒素血症组(LPS组)、右美托咪啶组(DEX组),每组10只。LPS组尾静脉注射脂多糖(LPS)5mg/kg,Dex组注射LPS后,静脉注射DEX7μg/kg,15 min后以5μg/kg·h持续泵注;S组按同样的方式泵注生理盐水。6h后取肺泡灌洗液和肺组织,ELISA检测肺泡灌洗液中TNF-a和IL-6含量;HE染色观察肺脏组织学变化;Realtime-PCR和Western-Blot法检测肺组织NF-κB、Myd88和TLR9 m RNA和蛋白表达水平。结果与S组相比,LPS组肺脏损伤明显,可见大量炎性细胞浸润,肺泡灌洗液中TNF-α和IL-6含量显著升高(<0.05),肺脏中NF-κB、Myd88和TLR9 m RNA和蛋白表达水平显著升高(<0.05)。经DEX干预后,肺脏损伤减轻,仅有少量炎性细胞,各指标表达量显著降低(<0.05)。结论 DEX抑制脓毒血症诱发ALI的早期炎症反应,可能与TLR9信号通路相关。展开更多
AIM:To investigate the regulation of mindin expression and the signaling pathway involved during inflammation.METHODS:C57BL/6 mice were treated with 3% dextran sulfate sodium (DSS) in drinking water for 6 d to induce ...AIM:To investigate the regulation of mindin expression and the signaling pathway involved during inflammation.METHODS:C57BL/6 mice were treated with 3% dextran sulfate sodium (DSS) in drinking water for 6 d to induce acute colitis,and then the colon was harvested for histological analysis or for RNA isolation.mRNA expression of mindin and nuclear factor (NF)-κB p65 was analyzed by quantitative real time polymerase chain reaction (RT-PCR) and mindin expression construct was conf irmed by Western blotting.Mouse macrophage and intestinal epithelial lineage cells were stimulated with different cytokines and toll-like receptor (TLR) ligands,before pNF-κB-luciferase activity was assessed using the Dual-Luciferase reporter assay system.RESULTS:mRNA expression of mindin was upregulated 4.7 ± 1.1 fold compared with the baseline during DSS-induced intestinal inflammation in the mice.Stimulation with CpG-ODN (a known TLR-9 ligand) induced 4.2 ± 0.3 fold upregulation of mindin expression in RAW 264.7 cells.Full-length of mindin was cloned from cDNA of mouse mesenteric lymph node,then the pCMV-Mindin-Flag expression vector was established and the protein expression level was confi rmed.Transfection of the mindin construct and stimulation with CpG-ODN signifi cantly increased the NF-κB-luciferase activity by 2.5 ± 0.3 and 4.5 ± 0.5 fold in RAW264.7 and CMT93 cells,respectively (P < 0.01).CONCLUSION:Mindin expression is upregulated during intestinal inflammation and may induce NF-κB promoter activation in a TLR-9 mediated manner.展开更多
目的:从形态学、免疫学、分子生物学角度探讨不同灸量治疗溃疡性结肠炎(UC)效果的异同。方法:32只SD大鼠随机分为空白组6只、模型复制组26只。采用三硝基苯甲酸/葡聚糖硫酸钠制备UC大鼠模型。模型复制成功后的大鼠按随机数字表分为模型...目的:从形态学、免疫学、分子生物学角度探讨不同灸量治疗溃疡性结肠炎(UC)效果的异同。方法:32只SD大鼠随机分为空白组6只、模型复制组26只。采用三硝基苯甲酸/葡聚糖硫酸钠制备UC大鼠模型。模型复制成功后的大鼠按随机数字表分为模型组、3壮组、6壮组、9壮组,每组各6只。各治疗组所取穴位为"天枢""大横",艾炷直接灸法,每次分别施灸3壮(3min)、6壮(6min)、9壮(9min),共治疗14次。观察大鼠治疗前后疾病活动指数(DAI),电镜、光镜观察结肠组织形态学改变,酶联免疫法检测大鼠血清中白介素-8(IL-8)、白介素-10(IL-10)含量,Western blot法检测大鼠结肠中Toll样受体9(TLR-9)和核转录因子-κB(NF-κB)p 65表达。结果:灸法可明显降低大鼠DAI的评分(与模型组比较均P<0.05);光镜与电镜结果显示,灸量越大,结肠组织腺体排列越规则。模型组血清IL-8含量升高,IL-10含量降低;与模型组比较,各治疗组IL-8降低,IL-10增高,其中9壮组和6壮组的变化较3壮组更明显(均P<0.05)。模型组结肠组织中TLR-9、NF-κB p 65大量表达;与模型组比较,各治疗组TLR-9、NF-κB p 65表达均降低,且以9壮组的变化最为明显(均P<0.05)。结论:艾灸可修复UC大鼠受损黏膜上皮,抑制血清中IL-8含量,提高血清中IL-10含量,通过抑制结肠组织中NF-κB p 65转录而下调TLR-9表达。灸量越大治疗效果越明显。展开更多
Toll-like receptors (TLRs) recognize specific motifs which are frequently present in bacteria, fungi, prokaryotes and viruses. Amongst TLRs, TLR9 can be activated by such bacterial or viral DNA fragments, immunoglobul...Toll-like receptors (TLRs) recognize specific motifs which are frequently present in bacteria, fungi, prokaryotes and viruses. Amongst TLRs, TLR9 can be activated by such bacterial or viral DNA fragments, immunoglobulin-DNA complexes or synthetic oligonucleotides, which all contain unmethylated cytosineguanine nucleotide sequences (CpGs). Emerging data indicate that TLR9 signaling has a role in, and may influence, colorectal carcinogenesis and colonic inflammation. CpGs are classified into three groups according to their influence on both the antigen-specific humoraland cellular immunity, and the production of type 1 interferons and proinflammatory cytokines. TLR9 activation via CpGs may serve as a new therapeutic target for several cancerous and various inflammatory conditions. Due to its probable anti-cancer effects, the application possibilities of TLR9-signaling modulation may be extremely diverse even in colorectal tumors. In this review we aimed to summarize the current knowledge about TLR-signaling in the pathogenesis and therapy of inflammatory bowel diseases and colorectal cancer. Due to the species-specific differences in TLR9 expression, however, one must be careful in translating the animal model data into the human system, because of the differences between CpG-oligodeoxynucleotide-responsive cells. TLR9 agonist DNA-based immunomodulatory sequences could also represent a promising therapeutic alternative in systemic inflammatory conditions and chronic colonic inflammations as their side effects are not significant.展开更多
文摘目的研究右美托咪啶(DEX)对内毒素血症大鼠急性肺损伤(ALI)的保护作用,探讨TLR9信号通路相关因子的表达差异。方法 SPF级雄性SD大鼠30只,随机分为3组:假手术组(S组)、内毒素血症组(LPS组)、右美托咪啶组(DEX组),每组10只。LPS组尾静脉注射脂多糖(LPS)5mg/kg,Dex组注射LPS后,静脉注射DEX7μg/kg,15 min后以5μg/kg·h持续泵注;S组按同样的方式泵注生理盐水。6h后取肺泡灌洗液和肺组织,ELISA检测肺泡灌洗液中TNF-a和IL-6含量;HE染色观察肺脏组织学变化;Realtime-PCR和Western-Blot法检测肺组织NF-κB、Myd88和TLR9 m RNA和蛋白表达水平。结果与S组相比,LPS组肺脏损伤明显,可见大量炎性细胞浸润,肺泡灌洗液中TNF-α和IL-6含量显著升高(<0.05),肺脏中NF-κB、Myd88和TLR9 m RNA和蛋白表达水平显著升高(<0.05)。经DEX干预后,肺脏损伤减轻,仅有少量炎性细胞,各指标表达量显著降低(<0.05)。结论 DEX抑制脓毒血症诱发ALI的早期炎症反应,可能与TLR9信号通路相关。
基金Supported by National Natural Science Foundation of China,No. 30750013
文摘AIM:To investigate the regulation of mindin expression and the signaling pathway involved during inflammation.METHODS:C57BL/6 mice were treated with 3% dextran sulfate sodium (DSS) in drinking water for 6 d to induce acute colitis,and then the colon was harvested for histological analysis or for RNA isolation.mRNA expression of mindin and nuclear factor (NF)-κB p65 was analyzed by quantitative real time polymerase chain reaction (RT-PCR) and mindin expression construct was conf irmed by Western blotting.Mouse macrophage and intestinal epithelial lineage cells were stimulated with different cytokines and toll-like receptor (TLR) ligands,before pNF-κB-luciferase activity was assessed using the Dual-Luciferase reporter assay system.RESULTS:mRNA expression of mindin was upregulated 4.7 ± 1.1 fold compared with the baseline during DSS-induced intestinal inflammation in the mice.Stimulation with CpG-ODN (a known TLR-9 ligand) induced 4.2 ± 0.3 fold upregulation of mindin expression in RAW 264.7 cells.Full-length of mindin was cloned from cDNA of mouse mesenteric lymph node,then the pCMV-Mindin-Flag expression vector was established and the protein expression level was confi rmed.Transfection of the mindin construct and stimulation with CpG-ODN signifi cantly increased the NF-κB-luciferase activity by 2.5 ± 0.3 and 4.5 ± 0.5 fold in RAW264.7 and CMT93 cells,respectively (P < 0.01).CONCLUSION:Mindin expression is upregulated during intestinal inflammation and may induce NF-κB promoter activation in a TLR-9 mediated manner.
文摘目的:从形态学、免疫学、分子生物学角度探讨不同灸量治疗溃疡性结肠炎(UC)效果的异同。方法:32只SD大鼠随机分为空白组6只、模型复制组26只。采用三硝基苯甲酸/葡聚糖硫酸钠制备UC大鼠模型。模型复制成功后的大鼠按随机数字表分为模型组、3壮组、6壮组、9壮组,每组各6只。各治疗组所取穴位为"天枢""大横",艾炷直接灸法,每次分别施灸3壮(3min)、6壮(6min)、9壮(9min),共治疗14次。观察大鼠治疗前后疾病活动指数(DAI),电镜、光镜观察结肠组织形态学改变,酶联免疫法检测大鼠血清中白介素-8(IL-8)、白介素-10(IL-10)含量,Western blot法检测大鼠结肠中Toll样受体9(TLR-9)和核转录因子-κB(NF-κB)p 65表达。结果:灸法可明显降低大鼠DAI的评分(与模型组比较均P<0.05);光镜与电镜结果显示,灸量越大,结肠组织腺体排列越规则。模型组血清IL-8含量升高,IL-10含量降低;与模型组比较,各治疗组IL-8降低,IL-10增高,其中9壮组和6壮组的变化较3壮组更明显(均P<0.05)。模型组结肠组织中TLR-9、NF-κB p 65大量表达;与模型组比较,各治疗组TLR-9、NF-κB p 65表达均降低,且以9壮组的变化最为明显(均P<0.05)。结论:艾灸可修复UC大鼠受损黏膜上皮,抑制血清中IL-8含量,提高血清中IL-10含量,通过抑制结肠组织中NF-κB p 65转录而下调TLR-9表达。灸量越大治疗效果越明显。
文摘Toll-like receptors (TLRs) recognize specific motifs which are frequently present in bacteria, fungi, prokaryotes and viruses. Amongst TLRs, TLR9 can be activated by such bacterial or viral DNA fragments, immunoglobulin-DNA complexes or synthetic oligonucleotides, which all contain unmethylated cytosineguanine nucleotide sequences (CpGs). Emerging data indicate that TLR9 signaling has a role in, and may influence, colorectal carcinogenesis and colonic inflammation. CpGs are classified into three groups according to their influence on both the antigen-specific humoraland cellular immunity, and the production of type 1 interferons and proinflammatory cytokines. TLR9 activation via CpGs may serve as a new therapeutic target for several cancerous and various inflammatory conditions. Due to its probable anti-cancer effects, the application possibilities of TLR9-signaling modulation may be extremely diverse even in colorectal tumors. In this review we aimed to summarize the current knowledge about TLR-signaling in the pathogenesis and therapy of inflammatory bowel diseases and colorectal cancer. Due to the species-specific differences in TLR9 expression, however, one must be careful in translating the animal model data into the human system, because of the differences between CpG-oligodeoxynucleotide-responsive cells. TLR9 agonist DNA-based immunomodulatory sequences could also represent a promising therapeutic alternative in systemic inflammatory conditions and chronic colonic inflammations as their side effects are not significant.
