腹膜间皮细胞过氧化物酶体增殖物活化受体γ的表达及其配体对CD40和细胞间黏附分子1的影响  被引量:3

Peroxisome proliferator-activated receptor γ(PPAR-γ) expresses in rat peritoneal mesothelial cells and PPAR-γ ligands down-regulate LPS-induced CD40 and ICAM-1 production

在线阅读下载全文

作  者:张云芳[1] 阳晓[1] 张亚杰[1] 孙玉玲[1] 孔庆瑜[1] 董秀清[1] 叶晓箐[1] 余学清[1] 

机构地区:[1]中山大学附属第一医院肾内科,广州510080

出  处:《中华肾脏病杂志》2005年第12期707-712,共6页Chinese Journal of Nephrology

基  金:国家自然科学基金(30271668);高等学校博士学科点专项科研基金(2002055805)

摘  要:目的 观察过氧化物酶体增殖物活化受体γ(PPAR-γ)在大鼠腹膜间皮细胞 (RPMCs)中的表达以及脂多糖(LPS)的调节作用,并探讨PPAR-γ天然配体15d-PGJ2及人工合 成配体cightazone对RPMCs表达CD40和ICAM-1的影响。方法 分离及培养RPMCs,常规传代 及鉴定,取第2代细胞用于实验研究。LPS不同浓度(0.1、1.0、10、50及100μg/ml)、LPS(1 μg/ml)处理后不同时间点及15d-PGJ2(3 μmol/L)、ciglitazone(10μmol/L)作用细胞36h后收 集细胞。RT-PCR检测PPAR-γ、CD40以及ICAM-1 mRNA表达。免疫细胞化学检测PPAR-γ在 RPMCs中的分布。Western印迹检测PPAR-γ及ICAM-1蛋白表达。结果 (1)常规培养的RPMCs 表达一定量PPAR-γ,表达部位主要分布于RPMCs细胞核内,在细胞浆微弱表达。(2)随着LPS 浓度逐渐增大,PPAR-γ蛋白表达水平呈逐渐增高的趋势,LPS浓度为10μg/ml时其表达为最 高峰。LPS(1μg/ml)作用12 h时PPAR-γ蛋白表达最强,PPAR-γ1表达高于PPAR-γ2;之后显 著降低,持续至72 h。(3)LPS刺激后RPMCs CD40mRNA表达显著增强;15d-PGJ2、ciglitazone显著 降低CD40 mRNA表达(P均<0.01)。(4)LPS刺激后RPMCs ICAM-1蛋白表达显著增加;15d-PGJ2 增加LPS介导的ICAM-1 mRNA表达(P<0.01),但显著抑制ICAM-1蛋白表达(P<0.05)。 ciglitazone对LPS介导的ICAM-1 mRNA和蛋白表达均有显著抑制作用(P均<0.05)。结论 RPMCs结构性表达PPAR-γ。LPS调节PPAR-γ表达呈现先增强后抑制趋势。PPAR-γ配体可显著 抑制LPS介导的CD40 mRNA和ICAM-1蛋白的表达。提示RPMCs功能性表达PPAR-γ,可能通 过负性调节炎症介质分泌而参与腹腔局部防御。Objective To detect the expression of PPAR-γ in rat peritoneal mesothelial cells (RPMCs) and to investigate the effect of PPAR-γ hgands, ciglitazone and 15-deoxy-△^12,14-prostaglandin J2 (15d-PGJ2) on the production of CD40 and ICAM-1 induced by lipopolysaccharide (LPS). Methods RPMCs were harvested from Sprague-Dawley rat peritoneal cavity and maintained under defined in vitro conditions. The cells were exposed respectively to different concentrations of LPS (0.1, 1.0,10,50 and 100 μg/ml) for 36 h or treated with LPS (1 μg/ml) from 0 h to 72 h. PPAR-γ mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR). The expression of PPAR-γ at the protein level and their intraeellular distribution was detected by Western blot and immunocytoehemistry respectively. Expression of ICAM-1 and CD40 on RPMCs under normal culture or stimulation respectively with LPS( 1 μg/ml), LPS+15d-PGJ2 ( 3 μmol/L), LPS+ciglitazone (10 μmol/L) and LPS+vehiele (DMSO 2 μl) was examined by RT-PCR and Western blot respectively. Results A single band with a predicted size (339 bp) was detected in RT-PCR products from RPMCs, which demonstrates PPAR-γ mRNA expression. Cultured RPMCs were further immunostained to confirm that specific signals of PPAR-γ were mainly localized in the nucleus with a weak staining in the cytoplasm. Stimulation of RPMCs with LPS resulted in a concentration-dependent increase in the protein level of PPAR-γ, with the peak at 10μg/ml. Treatment with 1 μg/ml LPS resulted in a time-dependent increase in the protein level of PPAR-γ, with the peak at 12 h. However, after that time point, the protein level of PPAR-γ was strongly attenuated by LPS. The expression of ICAM-1 mRNA and protein was upregulated significantly following stimulation with LPS (1 μg/ml). Both 15d-PGJ2 and eiglitazone significantly decreased the expression of ICAM-1 protein, but not mRNA. LPS increased the expression of CD40 mRNA significantly. Both 15d-P

关 键 词:过氧化物酶体激增剂 腹膜 CD40 细胞间黏附分子1 间皮细胞 

分 类 号:R329[医药卫生—人体解剖和组织胚胎学]

 

参考文献:

正在载入数据...

 

二级参考文献:

正在载入数据...

 

耦合文献:

正在载入数据...

 

引证文献:

正在载入数据...

 

二级引证文献:

正在载入数据...

 

同被引文献:

正在载入数据...

 

相关期刊文献:

正在载入数据...

相关的主题
相关的作者对象
相关的机构对象