羧甲基壳聚糖对体外培养人成纤维细胞自分泌生长因子及其形态结构的影响  

Effect of carboxymethylchitosan on autocrine growth factor and morphology of fibroblasts cultured in vitro

作　　者：肖海军[1] 侯春林[1] 刘亚平[1] 

机构地区：[1]第二军医大学长征医院骨科,上海200003

出　　处：《第二军医大学学报》2006年第8期848-852,共5页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(30170956)~~

摘　　要：目的:观察羧甲基壳聚糖对体外培养的人成纤维细胞形态结构及其分泌生长因子功能的影响,探讨其减轻创面过度愈合、防止术后粘连的可能机制。方法:体外培养的人成纤维细胞,经不同浓度(0.01、0.1、1.0、10 mg/ml)羧甲基壳聚糖作用4 d后,或经0.1 mg/ml羧甲基壳聚糖培养不同时间(1、2、3、4、56、d)后,采用间接酶联免疫吸附法(ELISA)和放射免疫法测定其自分泌转化生长因子-β1(TGF-β1)及表皮生长因子(EGF)的变化规律;并通过光镜及透射电镜观察不同浓度及不同作用时间后人成纤维细胞形态结构的变化。结果:羧甲基壳聚糖(≥0.1 mg/ml)可抑制成纤维细胞自分泌TGF-β1,且随浓度的增大及作用时间的延长而增强(P<0.05);但对EGF的自分泌无明显影响(P>0.05)。羧甲基壳聚糖(≥0.1 mg/ml)体外能够抑制成纤维细胞的增殖,引起超微结构的变化。结论:羧甲基壳聚糖浓度(≥0.1 mg/ml)体外可能通过改变人成纤维细胞超微结构,选择性抑制其自分泌TGF-β1,从而抑制细胞增殖,减轻组织粘连。Objective：To investigate the effect of carboxymethylchitosan on autocrine growth factor and morphology of fibroblasts cultured in vitro, so as to discuss the possible mechanism by which carboxymethylchitosan alleviates overhealing and prevents adhesion in wound healing. Methods： Fibroblasts were cultured in vitro. Fibroblasts of passage 4-6 were treated with different concentrations of carboxymethylchitosan （0.01,0.1,1.0 and 10 mg/ml） for 4 days or with 0. 1 mg/ml carboxymethylchitosan for 1, 2, 3, 4, 5, and 6 days. The levels of autocrine transforming growth factor-β1 （TGF-β1） and epidermal growth factor （EGF） of fibroblasts were determined by ELISA and radioimmunoassay. The fibroblastic morphology was detected by transmission electron microscopy （TEM） and microscope after fibroblasts were treated with different strategies. Results.. Carboxymethylchitosan（≥0.1 mg/ml）inhibited autocrine TGF-β1 of fibroblast in a time- and concentration dependent manner （P〈0.05）. However, Carboxymethylchitosan had no obvious influence on the secretion of EGF（P〉0.05）. Carboxymethylchitosan （≥0.1 mg/ml）also inhibited the proliferation of fibroblasts and caused their ultrastructural changes. Conclusion： Carboxymethylchitosan （≥0.1 mg/ml）can inhibit fibroblasts proliferation and reduce tissue adhesion, possibly through altering fibroblast ultrastructure and selectively inhibiting secretion of TGF-β1.

关 键 词：羧甲基壳聚糖 转化生长因子-Β1 表皮生长因子 自分泌 粘连 

分 类 号：R619[医药卫生—外科学]