兰州大学机构库 >学院待认领
过氧化物酶体增殖物激活受体-γ在慢性阻塞性肺疾病炎症中的作用
Alternative TitleEffects of Peroxisome Proliferator Activated Receptor-γ on the Inflammation in Chronic Obstructive Pulmonary Disease
曾晓丽
Thesis Advisor刘晓菊
2011-05-17
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword慢性阻塞性肺疾病 外周血单个核细胞 罗格列酮 GW9662 炎症 过氧化物酶体增殖物激活受体-γ 核因子-κB 肿瘤坏死因子-α
Abstract目的 观察过氧化物酶体增殖物激活受体-γ (PPAR-γ)激动剂罗格列酮及PPAR-γ拮抗剂GW9662对慢性阻塞性肺疾病(COPD)患者PPAR-γ、核因子-κB (NF-κB)和肿瘤坏死因子-α (TNF-α)的影响,探讨PPAR-γ在COPD炎症中的作用。 方法 选择COPD急性加重期患者30例、健康体检者24例为研究对象,分离外周血单个核细胞(PBMCs),并用罗格列酮和GW9662对COPD患者PBMCs进行干预。实验分为4组:健康对照组(A组)、COPD组(B组)、COPD罗格列酮干预组(C组)及COPD罗格列酮联合GW9662干预组(D组),采用实时荧光定量PCR (Real-Time PCR)检测各组PBMCs中PPAR-γ和NF-κB mRNA的表达,细胞免疫荧光法结合激光扫描共聚焦显微镜检测PPAR-γ和NF-κB蛋白表达及核转位,酶联免疫吸附试验(ELISA)检测细胞培养上清液中TNF-α的含量。 结果 (1)B组PPAR-γ mRNA和蛋白表达(0.52±0.10、55.12±10.83)分别低于A组(1、84.66±9.48) (P均<0.05);NF-κB mRNA和蛋白的表达(1.69±0.07、145.40 ±17.14)分别高于A组(1、117.55±6.71) (P分别<0.01、<0.05);TNF-α的浓度(96.20±1.44)μg/L明显高于A组(85.33±1.03)μg/L (P<0.01);B组PPAR-γ蛋白主要位于细胞浆,NF-κB蛋白主要位于细胞核,而A组PPAR-γ、NF-κB蛋白在细胞浆和细胞核中均有表达。(2)经罗格列酮干预后,PPAR-γ mRNA和蛋白表达(4.47±0.11、204.36±11.89)较B组明显增高(P均<0.01);NF-κB mRNA和蛋白的表达(0.33±0.04、58.97±13.69)较B组明显降低(P均<0.01);TNF-α的浓度(63.04±2.54)μg/L较B组明显降低(P<0.01);PPAR-γ蛋白转位于细胞核,NF-κB蛋白转位于细胞浆。(3)用GW9662预处理后再给予罗格列酮干预,PPAR-γ mRNA和蛋白表达(2.25±0.31、141.59±23.44)较C组明显降低(P均<0.01),但高于B组和A组(P均<0.01);NF-κB mRNA和蛋白的表达(0.64±0.02、90.06±9.65)较C组增高(P分别<0.01、<0.05),但低于B组和A组(P<0.01或<0.05);TNF-α的浓度(83.27±1.85)μg/L较C组明显升高(P<0.01),但低于B组(P<0.01);PPAR-γ蛋白又转回细胞浆,NF-κB蛋白部分转移至细胞核。(4) PPAR-γ蛋白的表达与NF-κB蛋白的表达和TNF-α的浓度均呈负相关(P均<0.01),NF-κB蛋白表达与TNF-α的浓度呈正相关(P<0.01)。 结论 COPD的炎症可能与PPAR-γ的表达及活性不足有关;罗格列酮能通过上调PPAR-γ的表达及促进其核转位来抑制NF-κB的表达及核转位,进而抑制TNF-α的分泌,在COPD炎症中起重要的作用。
Other AbstractObjective To study the effects of peroxisome proliferator activated receptor-γ (PPAR-γ) agonist rosiglitazone and PPAR-γ antagonist GW9662 on PPAR-γ, nuclear factor–κB (NF-κB) and tumor necrosis factor-α (TNF-a), so as to explore the effects of PPAR-γ on the inflammation in chronic obstructive pulmonary disease (COPD). Methods Thirty patients of COPD and 24 healthy controls were included. The peripheral blood mononuclear cells (PBMCs) were isolated from blood of the patients with COPD and healthy controls. The PBMCs with COPD were theated with rosiglitazone and GW9662. The PBMCs were divided into 4 groups: control group (group A), COPD group (group B), COPD rosiglitazone group (group C), COPD rosiglitazone and GW9662 group (group D). The expression of PPAR-γ mRNA and NF-κB mRNA was measured with Real-Time PCR. The expression of PPAR-γ and NF-κB protein and nuclear translocation were detected using immunofluorescence with laser scanning confocal microscopy. The TNF-a level in cultural supernatant was measured with ELISA. Results (1) The mRNA and protein levels of PPAR-γ were lower in group B (0.52±0.10, 55.12±10.38) than that in group A (1, 84.66±9.48) (P all < 0.05), while the levels of NF-κB mRNA and protein were higher in group B (1.69±0.07, 145.40±17.14) than that in group A (1, 117.55±6.71) (P respectively < 0.01, < 0.05). TNF-a level was significantly higher in group B (96.20±1.44)μg/L than that in group A (85.33±1.03)μg/L (P < 0.01). The proteins of PPAR-γ and NF-κB were respectively located in cytoplasm and in nucleus in group B, meanwhile the proteins of PPAR-γ and NF-κB were located in both cytoplasm and nucleus in group A. (2) After the treatment of rosiglitazone (group C), the mRNA and protein levels of PPAR-γ (4.47±0.11, 204.36±11.89) were significantly increased compared with group B (P all < 0.01), while the mRNA and protein levels of NF-κB (0.33±0.04, 58.97±13.69) were remarkably decreased compared with group B (P all < 0.01). TNF-a level (63.04±2.54)μg/L in group C was significantly lower than that in group B (P < 0.01). PPAR-γ protein was translocated from cytoplasm into nucleus and NF-κB protein was translocated from nucleus into cytoplasm. (3) Before rosiglitazone treatment with GW9662 pretreated (group D), the mRNA and protein levels of PPAR-γ (2.25±0.31, 141.59±23.44) were significantly decreased compared to group C (P all < 0.01), but higher compared with group B and group A (P all < 0.0...
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Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/223352
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
曾晓丽. 过氧化物酶体增殖物激活受体-γ在慢性阻塞性肺疾病炎症中的作用[D]. 兰州. 兰州大学,2011.
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