槟榔提取物抑制人类口腔粘膜角朊细胞生长的实验研究  

Research on Inhibition of Areca Nut Extract on Human Buccal Keratinocytes

作　　者：冯云枝[1] 凌天牖[1] 

机构地区：[1]湖南医科大学附属第二医院口腔科,410011

出　　处：《华西口腔医学杂志》1999年第3期233-235,I011,共4页West China Journal of Stomatology

摘　　要：目的 :建立人类口腔粘膜上皮角朊细胞体外原代培养方法 ;探讨口腔粘膜下纤维性变 (OSF)患者上皮厚度改变的机理。方法 :先对人类口腔粘膜上皮角朊细胞 (KC)进行分离、培养和鉴定 ,然后用四唑盐 (MTT)比色试验检测 OSF患者和正常人口腔粘膜上皮 KC增殖状况 ,并且观察槟榔提取物 (ANE)对 KC增殖的影响。结果 :体外培养的 KC呈多边形 ,胞浆内有丰富的张力细丝 ,表面有大量微绒毛 ,角蛋白抗体免疫组化染色呈阳性。且体外培养的OSF患者 KC与正常对照组 KC增殖无明显差异 ,ANE以浓度 -效应依赖关系抑制 KC增殖。结论 :用 dispase消化培养法可成功地培养出纯净的 KC;ANE对口腔粘膜 KC有细胞毒作用。Objective: To culture human oral mucosa epithelial cells in vitro and study the mechanism of changes in the epithelia of oral submucous fibrosis (OSF). Methods: The keratinocytes were obtained from normal buccal mucosa (NM KC) and OSF buccal mucosa (OSF KC) with dispase trypsine and cultured in vitro. Then the cell proliferation of keratinocytes incubated with or without areca nut extract in the presence of 10% fetal calf serum for 48 hours at 37℃ in 5% CO\-2 and air were monitered by use of thiazolyl blue (MTT) assay. Results: Primary culture grew in small islands that coalesced at confluency. Ultrastructually the cells contained a large of tonofilament and microvilli. Immunohistochemistry demonstrated uniform staining of the cells with antibodies to keratins. There was no difference in cell proliferation between OSF KC and NM KC, and areca nut extract inhibited keratinocyte proliferation in a concentration dependant manner. Conclusion: The results suggested that utilizing dispase Ⅱ can establish fibroblast free culture for growing of human oral mucosa keratinocytes and areca nut extract is a cytotoxic agent to human buccal keratinocytes.

关 键 词：角朊细胞 槟榔提取物 口腔粘膜 细胞增殖 OSF 

分 类 号：R781.502[医药卫生—口腔医学]