人腹膜间皮细胞的培养及特征  

Culture and characterization of human peritoneal mesothelial cells

作　　者：刘伏友[1] 段绍斌[1] 龙志高[2] 肖平[1] 陈星[1] 潘乾[2] 罗季安[1] 彭佑铭[1] 

机构地区：[1]中南大学湘雅二医院肾内科,长沙410011 [2]中南大学中国医学遗传国家重点实验室,长沙410078

出　　处：《湖南医科大学学报》2001年第4期321-324,共4页Bulletin of Hunan Medical University

基　　金：卫生部临床重点项目资助 ( 2 19977191)

摘　　要：目的 :建立人腹膜间皮细胞 (HPMC)培养模型 ,检测HPMC细胞外基质蛋白及白细胞介素 (IL - 8)的表达。方法 :采用胰蛋白酶 -EDTA消化法 ,从人的腹膜组织中分离HPMC。采用形态学和链霉菌抗生物素蛋白 过氧化物酶连接法 (即SP法 ) ,对培养细胞进行鉴定。采用SP法检测HPMC纤维连接蛋白 (FN)、胶原Ⅲ和胶原V及转化生长因子 (TGF) β1表达。采用逆转录多聚酶链反应 (RT -PCR)检测HPMCIL - 8mRNA和FNmRNA的表达。结果 :光镜示细胞融合后呈多边形 ,超微结构下可见丰富的微绒毛和内质网 ,免疫组化染色结果示HPMC可表达角蛋白 ,波形蛋白 ,胶原Ⅲ ,FN ,TGFβ1蛋白 ,Ⅷ因子、胶原V表达阴性。人腹膜间皮细胞亦表达FNmRNA和IL - 8mRNA。结论 :HPMC培养模型的建立 ,可为进一步研究腹膜纤维化的防治及IL - 8在腹膜透析中的作用提供理论依据。Objective To establish a reproducible model for culture of human peritoneal mesothelial cells (HPMCs), and observe the expressions of extracellular matrix (ECM) protein and interleukin-8 (IL-8) in HPMC. Methods Mesothelial cells were isolated from human omenta by trypsin EDTA disaggregation. HPMCs were identified by morphology and streptomyces protein-peroxidase (SP) method. In the same way, expressions of fibronectin, collagen type Ⅲ, V and transforming growth factor β 1 (TGFβ 1) were measured. Expression of IL-8 mRNA and FN mRNA in the HPMCs was detected by reverse transcription (RT) and polymerase chain reaction (PCR) amplification. Results Confluent HPMCs appeared multipolar under microscope; numerous surface microvilli and an abundant endoplasmic reticulum were observed. Cultured HPMCs coexpressed cytokeratin and vimentin, and synthesized fibronectin, TGFβ 1 and collagen types Ⅲ (instead of type V). Expressions of IL-8 mRNA and FN mRNA were also observed in HPMCs. Conclusion Establishment of cultured HPMCs model will provide the basis of the research in preventment of peritoneal fibrosis and roles of IL-8 in peritoneal dialysis.

关 键 词：腹膜间皮细胞 基质 转化生长因子Β1 白细胞介素-8 细胞培养 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]