摘要
目的 观察cathepsinK在脂肪细胞分化中的表达趋势，初步探讨其在脂肪细胞分化中的作用。方法 4～6周SD大鼠，酶消化法获得前脂肪细胞，黄嘌呤、胰岛素、地塞米松方案进行诱导，RT-PCR和Western blot方法检测cathepsinK在脂肪细胞分化过程中的表达。用不同浓度的cathepsinK抑制剂E-64进行干预，油红染色评估效果。结果 前脂肪细胞汇合后经异丁基-甲基-黄嘌呤、地塞米松、胰岛素诱导，60％～70％的细胞分化为成熟的脂肪细胞表型，且与脂肪细胞分化紧密相关的转录因子——过氧化物增生激活物受体（PPAR-1）持续表达，可用于实验研究。在脂肪细胞分化的过程中cathepsin K mRNA和蛋白表达上升（P〈0．05）。加入不同浓度的E-64后，脂肪细胞分化进程受到抑制，且呈剂量依赖关系。结论 脂肪细胞分化前后cathepsin K的表达存在差异，其抑制剂可以阻止脂肪细胞分化。提示cathepsin K在脂肪细胞分化过程中发挥作用。
Objective To observe the tendency of cathepsin K expression during adipocyte differentiation, and to study its effects. Methods White fat precursor ceils were isolated from the epididymal fat pad of 4-6 week-old male SD rats. Ceils were then induced to differentiate in the presence of DMEM/F12 media supplemented with insulin, dexamethasone （DX）, isobutyl- methylxanthine （IBMX）. RT-PCR and Western blot were used to detect the gene/protein expression of cathepsin K. Through cathepsin K depressor E-64 intervention in different concentrations, oil red O staining was applied to detect the degree of preadipocytes differentiation. Results About 60%-70% primary rat preadipocytes were induced to differentiate using IBMX, insulin, and DX. They also promoted the expressions of critical adipogenic transcription factors, peroxisome proliferator-activated receptor 3＇ （PPAR-3＇）. Expressions of cathepsin K mRNA and protein were upregulated during adipocyte differentiation. While intervention with E-64, the process was inhibited dose-dependently. Conclusion Levels of cathepsin K were variable during adipocyte differentiation. Its depressor E-64 could inhibit the process. These data suggest that cathepsin K plays a role in adipocyte differentiation.
出处
《上海交通大学学报（医学版）》
CAS
CSCD
北大核心
2006年第1期59-62,共4页
Journal of Shanghai Jiao tong University:Medical Science
基金
国家自然科学基金(30100085)资助项目
