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

doi:10.3969/j.issn.1674-5817.2014.02.008

Abstract

Abstract: 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 µmol/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.

Key words: NaF, Chondrocytes, GAG, II type collagen

CLC Number:

Q95-33

WANG Ning, FAN Yu-lan, SHI Zhi-chong, ZHENG Wei-dong, LU Wei, XIAO Ping. Effects of Excess Fluoride on Secretion of GAG and II Type Collagen in vitro Chondrocytes of Rats[J]. Laboratory Animal and Comparative Medicine, 2014, 34(2): 120-125.

References

[1] Bely M, Ferenez G.Experimental osteofluorosis and anthrofluorosis in rats[J]. Clin Rheumatol, 1996, 15:529-530.
[2] Meng H, Zhang T, Liu W, et al.Sodium fluoride induces apoptosis through the downregulation of hypoxia-inducible factor-1α in primary cultured rat chondrocytes[J]. Int J Mol Med, 2014, 33(2):351-358.
[3] Nishida T, Kubota S, Aoyama E,et al.Impaired glycolytic metabolism causes chondrocyte hypertrophy-like changes via promotion of phospho-Smad1/5/8 translocation into nucleus[J]. Osteoarthritis Cartilage, 2013, 21(5):700-709.
[4] 司徒镇强, 吴志英. 细胞培养[M]. 西安: 世界图书出版社, 1996:187-190.
[5] Chen JT, Liang JB, Chou CL, et al.Glucosamine sulfate inhibits TNF-alpha and IFN-gamma-induced production of ICAM-1 in human retinal pigment epithelial cells in vitro[J]. Invest Ophthalmol Vis Sci, 2006, 47(2):664-672.
[6] Guo X, Wu S, He Y, et al.Effect of subchronic fluoride exposure on pathologic change and beta-catenin expression in rat bone tissue[J]. Wei Sheng Yan Jiu, 2011, 40(3):304-307.
[7] 肖萍, 董妙珠, 叶于薇, 等. 不同月龄豚鼠原发性骨关节炎的病理变化[J]. 环境与职业医学, 2004, 21(6):460-462.
[8] Machoy-Mokrzyńska A.Fluorine as a factor in premature aging[J]. Ann Acad Med Stetin,2004,50(Suppl 1):9-13.
[9] Claassen H, Cellarius C, Scholz-Ahrens KE, et al.Extracellular matrix changes in knee joint cartilage following bone-active drug treatment[J].Cell Tissue Res, 2006, 324(2):279-289.
[10] 徐鹏, 郭雄. 过量氟化物对软骨损伤的研究进展[J]. 国外医学: 医学地理分册, 2001, 22(1):1-3.
[11] 朴春吉, 侯立中, 杨同书, 等. 过量氟化物对软骨基质胶原蛋白代谢的影响[J]. 中国地方病防治杂志, 1996, 11:75-78.
[12] Stoop R, Buma P, Van der Kraan PM, et al. Type II collagen degradation in articular cartilage fibrillation after anterior cruciate ligament transaction in rats[J]. Osteoarthritis Cartilage, 2001, 9:308-315.
[13] Gibson GJ, Verner JJ, Nelson FR, et al.Degradation of the cartilage collagen matrix associated with changes in chondrocytes in osteoathrosis: assessment by loss of background fluorescence and immunodetection of matrix components[J]. J Orthop Res, 2001, 19:33-42.
[14] Yasuhara R, Miyamoto Y, Akaike T, et al.Interleukin-1beta induces death in chondrocyte-like ATDC5 cells through mitochondrial dysfunction and energy depletion in a reactive nitrogen and oxygen species-dependent manner[J].Biochem J, 2005, 389(Pt 2):315-323.