胰岛素在氟诱导成骨作用中的地位  

The status of insulin in the role of fluoride-induced osteogenesis

作　　者：张秀云[1] 王岩[1] 赵志涛[1] 杨晨[1] 徐辉[1] 

机构地区：[1]吉林大学药学院,长春130021

出　　处：《中国科技论文》2015年第18期2121-2126,共6页China Sciencepaper

基　　金：高等学校博士学科点专项科研基金资助项目(20130061110084);吉林大学白求恩计划(B)资助项目(2012222)

摘　　要：采用实时PCR测试了投氟大鼠的骨组织成骨和胰岛素受体等基因表达情况,采用免疫组化技术观察了胰岛分泌胰岛素情况,以及采用生化方法检测血清中的胰岛素、血糖和血脂等变化,利用体外成骨细胞染氟考察了胰岛素受体在成骨细胞内表达及氟联合胰岛素对骨髓间充质干细胞的成骨作用影响。结果表明:给予低剂量氟3个月后,大鼠骨组织碱性磷酸酶、骨钙素、Runx2(Runt相关转录因子)及胰岛素受体表达显著提高,而高氟组成骨标志基因碱性性磷酸酶、骨钙素、Runx2也明显提高,说明给予大鼠氟3个月能够诱导成骨;给与低剂量氟从1个月到3个月后,大鼠血清的胰岛素浓度明显提高,高氟作用3月后,胰岛素分泌降低,这与高剂量氟给氟3月大鼠胰岛素受体表达明显下降相一致。体外实验中,ρ(F-)=2mg/L和8mg/L氟明显促进了成骨细胞内抗胰岛素受体荧光抗体的强度,而且10μg/mL胰岛素联合ρ(F-)=1mg/L和4mg/L能够刺激成骨细胞活性。氟ρ(F-)=1mg/L和4mg/L刺激了成骨细胞碱性磷酸酶的活性。体内氟中毒大鼠模型实验结果表明,低剂量氟作用3个月刺激成骨作用的同时,伴随着胰岛素分泌增强,并促进成骨细胞内胰岛素受体表达增多,体外实验也表明了胰岛素刺激成骨细胞增生的作用,这些实验数据说明胰岛素参与了低剂量氟诱导成骨作用机制。The gene expression of osteogenesis and insulin receptor for the rats exposed to fluoride was analyzed by realtime PCR.The insulin secretion by pancreas islet was observed by immunohistochemistry,and then the levels of insulin,blood glucose and serum fat were tested by means of biochemical methods.The effects of insulin receptor expression in osteoblasts and fluorine in combination with insulin on bone marrow mesenchymal stem cells into bone were studied by in vitro osteoblasts infected by fluorine.The results show that expression of Alkaline phosphatase（ALP）,osteocalcin（OCN）,Runt-related transcription factor 2（Runx2）and Insulin receptor（InR）increase after low-dose fluoride administration for 3months and high-dose fluoride also stimulate the expression of ALP,OCN and Runx2,which suggest the osteogenesis action of low and high dose of fluoride.The insulin level obviously enhances in rats treated with low-dose fluoride from 1month to 3months;however,it reduces after high-dose fluoride administration for 3months,which is consistent with the expression of InR in rats after high-dose fluoride administration for 3months.In vitro studies indicateρ（F-）= 2mg/L and 8mg/L stimulate the protein expression of insulin receptor by immunofluorescence technique with anti-InR antibody.The 10μg/mL insulin and co-treatment fluoride promote cell viability of bone marrow stromal cells,and single insulin administration significantly enhances cell viability.The fluoride ofρ（F-）=1mg/L and4mg/L markedly stimulates ALP activity.These data of fluorosis animal model show low-dose fluoride induced osteogenesis and insulin secretion and increased InR expression in rats,and which implies the role of insulin on the mechanism underlying the osteogenesis induced by fluoride.In vitro cell culture also indicates that insulin stimulates cell osteoblastic cell proliferation.All these results suggest the insulin is involved in the mechanism underlying low-dose of fluoride induced osteogenesis action.

关 键 词：地方病学 氟骨症 成骨作用 胰岛素 

分 类 号：R599.1[医药卫生—内科学]