CAJ | 학술논문

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。
목적：탐토봉극련접통신증강제전렬선소E2대지방세포향성골세포횡향분화급세포간봉극련접통신적영향。방법선취3월령신서란대백토복고구처지방조직，분리제취성숙지방세포，사기거분화득도거분화지방세포，취제삼대거분화적지방세포진행성골유도，가입GJIC증강제PGE2설위증강조，병설립대조조，MTT법관찰PGE2대세포배증시부유영향，진행천소홍개결절염색화I형효원면역조화염색대성골세포정성검측，Western blot기술화화흔표기염료시종법검측봉극련접단백43（Cx43）적표체。결과 MTT법검측결과현시PGE2대세포생장증식무현저영향。천소홍염색현시균발현자홍색결절。면역조화염색현시증강조I형효원표체증강，SLDT법현시증강조형광염료확산고우대조조，Western blot기술검측결과현시증강조Cx43단백적표체증고。결론 PGE2가이증강봉극련접통신이증강향성골세포분화적능력。
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.