GJIC增强剂PGE_2对兔脂肪细胞向成骨细胞横向分化的影响  

The effect of GJIC stimulating agent PGE2 on the osteoblast transdifferentiation by adipocytes in rabbits

作　　者：王吉博[1] 彭浩[1] 王凤宇[1] 齐新文[1] 王兆杰[1] 

机构地区：[1]遵义医学院珠海校区手术学教研室,珠海519041

出　　处：《中国骨质疏松杂志》2014年第9期1079-1083,共5页Chinese Journal of Osteoporosis

基　　金：贵州省科技厅基金资助项目[黔科合J字(2007)2227号];遵义医学院博士基金资助项目(遵医F-129)

摘　　要：目的探讨缝隙连接通讯增强剂前列腺素E2对脂肪细胞向成骨细胞横向分化及细胞间缝隙连接通讯的影响。方法选取3月龄新西兰大白兔腹股沟处脂肪组织,分离提取成熟脂肪细胞,使其去分化得到去分化脂肪细胞,取第三代去分化的脂肪细胞进行成骨诱导,加入GJIC增强剂PGE2设为增强组,并设立对照组,MTT法观察PGE2对细胞倍增是否有影响,进行茜素红钙结节染色和I型胶原免疫组化染色对成骨细胞定性检测,Western blot技术和划痕标记染料示踪法检测缝隙连接蛋白43(Cx43)的表达。结果 MTT法检测结果显示PGE2对细胞生长增殖无显著影响。茜素红染色显示均发现紫红色结节。免疫组化染色显示增强组I型胶原表达增强,SLDT法显示增强组荧光染料扩散高于对照组,Western blot技术检测结果显示增强组Cx43蛋白的表达增高。结论 PGE2可以增强缝隙连接通讯以增强向成骨细胞分化的能力。Objective To explore the effect of gap junction strengthening agent PGE2 on the osteoblast transdifferentiation by adipocytes and the gap junction intercellular communication （ GJIC） .Methods Groin adipose tissue was taken from 3-month-old New Zealand white rabbits and the mature adipocytes were collected.The mature adipocytes were dedifferentiated and the dedifferentiated adipocytes were obtained.The 3rd generation of the dedifferentiated adipocytes were induced and differentiated into osteoblasts.PGE2 was additioned in the enhanced group.The control group was also established.The effect of PGE2 on osteoblast proliferation was detected using MTT assay.Osteoblast phenotype was examined using Alizarin red staining and collagen type I immunohistochemistry staining.The expression of Cx43 was measured using Western blotting and sorape-loading and dye transfet （ SLDT） method.Results The results of MTT assay showed that PGE2 had no significant effect on cell proliferation.The results of Alizarin red staining showed that calcified nodules were purple stained in all groups. The results of collagen type I immunohistochemistry staining showed that the expression of collagen type I was stimulated in the enhanced group.The results of SLDT showed that the spread of fluorescent dye was more in the enhanced group than in the control group.The results of Western blotting showed that the expression of Cx43 was stimulated in the enhanced group.Conclusion PGE2 enhanced the capability of osteoblast transdifferentiation by enhancement of GJIC.

关 键 词：脂肪细胞 成骨细胞 去分化 缝隙连接通讯 

分 类 号：R68[医药卫生—骨科学]