摘要
目的探讨过量氟对体外培养软骨细胞糖胺多糖(glycosaminoglycan,GAG)和胶原分泌的影响。方法取2月龄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,分空白对照组和NaF受试组(6.25μmol/ml、12.5μmol/ml、25μmol/ml、50μmol/ml和100μmol/ml),孵育6 d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6 d后,大鼠软骨细胞培养液GAG含量各组比较,差异无统计学意义(P>0.05)。阿利新蓝染色显示NaF处理大鼠软骨细胞后,细胞内GAG分泌颗粒无明显差异;Ⅱ型胶原免疫组化染色则显示,过量氟可引起软骨细胞外Ⅱ型胶原的分泌增多,半定量分析显示随着剂量升高,软骨细胞外Ⅱ型胶原的分泌逐渐增多。结论 NaF对大鼠软骨细胞GAG分泌无影响,但可引起Ⅱ型胶原分泌增多,有一定的剂量-反应关系。
Objective To observe the effect of excess fluoride in vitro on secretion of GAG and Ⅱtype collagen of rat chondrocytes.Methods Chondrocytes were separated from the 2-month old rats and cultured in the cell bottles.Then the chondrocytes were exposed to various concentrations of NaF,i.e,0,6.25,12.5,25,50 and 100 μmol/ml.After 6 days＇ culture,GAG contents were measured and Alcian Blue dye,DMB dye and Ⅱtype collagen immunohistochemistry dye were also done to measure the secretion of GAG and Ⅱtype collagen.Results After 6 days of treatment,there was no significant difference of GAG contents of cell culture medium among NaF treated groups and control group（P 〉0.05）.Alcian Blue dye and DMB dye showed that secretion of GAG in all NaF treated groups did not change compared with the control group.However,the immunohistochemistry dye showed the Ⅱtype collagen gradually increased with increment of NaF concentration.Conclusion NaF did not induce change of the secretion of GAG,but increment of Ⅱtype collagen of rat chondrocytes in vitro.
出处
《职业卫生与应急救援》
2010年第5期239-242,共4页
Occupational Health and Emergency Rescue
基金
上海市卫生局科研课题局级科研项目(2007180)
