氟浓度对体外培养大鼠软骨细胞糖胺多糖和Ⅱ型胶原分泌的影响

Effects of Excess Fluoride on Secretion of GAG and Ⅱ Type Collagen in vitro Chondrocytes of Rats

目的 探讨氟浓度对体外培养软骨细胞糖胺多糖（GAG）和Ⅱ型胶原分泌的影响.方法 取2月龄雌性SD大鼠下肢髋关节及膝关节软骨,分离、培养出软骨细胞后,接种于细胞培养瓶,经鉴定后,分为空白对照组和NaF受试组（6.25 μmol/ml、12.50μmol/ml、25.00 tmol/ml、50.00 μmol/ml和100.00 μmol/ml）,孵育6d后,分别进行GAG含量测定、阿利新蓝染色和Ⅱ型胶原免疫组化染色.结果 NaF受试6d后,大鼠软骨细胞培养液GAG含量各组无显著性差异（P＞0.05）;阿利新蓝染色显示,大鼠软骨细胞内的糖原异染颗粒亦无明显差异;Ⅱ型胶原免疫组化染色则显示,不同剂量NaF可引起对软骨细胞外Ⅱ型胶原染色积分随剂量的升高逐渐上升（P＜0.05）.结论 NaF对大鼠软骨细胞GAG分泌可能无影响,但可能影响Ⅱ型胶原的分泌.

Objective To observe the effect of secretion of GAG and II type collagen of excess fluoride on in vitro chondrocytes of rats. Methods Chondrocytes of rats was separated from the 2-month old female rats, and cultured in the cell bottles. After identification, six groups were designed in the research including control group and NaF group （6.25 μmol/ml, 12.50 μmol/ml, 25.00 μmol/ml, 50.00 pmol/ml and 100.00 μmol/ml）. After 6 days of administration, GAG contents were measured. Alcian Blue dye and II type collagen immunohistochemistry dye were also performed on the in vitro chondrocytes of rats. Results After 6 days of treatment, there were no significant difference of GAG contents of cell culture medium between NaF groups and control group （P〉0.05）. Alcian Blue dye showed that secretion of GAG content of all groups did not change after administered by NaF. But II type collagen immunohistochemistry dye showed that scores of II type collagen immunohistochemistry dye increased according to dosage of NaF leveled up （P〈0.05）. Conclusions NaF may not affect the content of GAG secreted by the chondrocytes of rats in vitro, but can affect the secretion of II type collagen.