中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2015年
5期
888-893
,共6页
唐照%张叶敏%李明欣%汪长华
唐照%張葉敏%李明訢%汪長華
당조%장협민%리명흔%왕장화
泛素蛋白酶系统%过氧化物酶体增殖物活化受体γ%热休克蛋白90%脂肪细胞%细胞分化
汎素蛋白酶繫統%過氧化物酶體增殖物活化受體γ%熱休剋蛋白90%脂肪細胞%細胞分化
범소단백매계통%과양화물매체증식물활화수체γ%열휴극단백90%지방세포%세포분화
Ubiquitin-proteasome system%Peroxisome proliferator-activated receptor gamma%Heat shock pro-tein 90%Adipocytes%Cell differentiation
目的：探讨泛素蛋白酶系统（ ubiquitin-proteasome system，UPS）在体外脂肪细胞分化中的作用。方法：常规方法诱导前脂肪细胞分化为脂肪细胞，Western blot检测蛋白质表达，免疫共沉淀检查蛋白质间结合，油红O染色检测脂肪细胞中的脂质，RT-PCR检测mRNA表达。结果：UPS抑制剂硼替佐米可抑制3T3-L1前脂肪细胞分化为脂肪细胞，表现为细胞内脂质含量降低以及脂肪细胞标志蛋白mRNA表达的降低。蛋白激酶G激动剂西地那非可激活UPS，并增强脂肪细胞的分化。抑制UPS可降低热休克蛋白90（ HSP90）和过氧化物酶体增殖物活化受体γ（ PPARγ）间的结合；同时降低细胞可溶性组分中HSP90和PPARγ的表达，增强其在不可溶性组分中的表达。HSP90特异性的N末端抑制剂格尔德霉素可抑制西地那非促进的PPARγ表达和脂肪细胞的分化。结论：泛素蛋白酶系统通过HSP90调节PPARγ的表达，从而影响脂肪细胞的分化。
目的：探討汎素蛋白酶繫統（ ubiquitin-proteasome system，UPS）在體外脂肪細胞分化中的作用。方法：常規方法誘導前脂肪細胞分化為脂肪細胞，Western blot檢測蛋白質錶達，免疫共沉澱檢查蛋白質間結閤，油紅O染色檢測脂肪細胞中的脂質，RT-PCR檢測mRNA錶達。結果：UPS抑製劑硼替佐米可抑製3T3-L1前脂肪細胞分化為脂肪細胞，錶現為細胞內脂質含量降低以及脂肪細胞標誌蛋白mRNA錶達的降低。蛋白激酶G激動劑西地那非可激活UPS，併增彊脂肪細胞的分化。抑製UPS可降低熱休剋蛋白90（ HSP90）和過氧化物酶體增殖物活化受體γ（ PPARγ）間的結閤；同時降低細胞可溶性組分中HSP90和PPARγ的錶達，增彊其在不可溶性組分中的錶達。HSP90特異性的N末耑抑製劑格爾德黴素可抑製西地那非促進的PPARγ錶達和脂肪細胞的分化。結論：汎素蛋白酶繫統通過HSP90調節PPARγ的錶達，從而影響脂肪細胞的分化。
목적：탐토범소단백매계통（ ubiquitin-proteasome system，UPS）재체외지방세포분화중적작용。방법：상규방법유도전지방세포분화위지방세포，Western blot검측단백질표체，면역공침정검사단백질간결합，유홍O염색검측지방세포중적지질，RT-PCR검측mRNA표체。결과：UPS억제제붕체좌미가억제3T3-L1전지방세포분화위지방세포，표현위세포내지질함량강저이급지방세포표지단백mRNA표체적강저。단백격매G격동제서지나비가격활UPS，병증강지방세포적분화。억제UPS가강저열휴극단백90（ HSP90）화과양화물매체증식물활화수체γ（ PPARγ）간적결합；동시강저세포가용성조분중HSP90화PPARγ적표체，증강기재불가용성조분중적표체。HSP90특이성적N말단억제제격이덕매소가억제서지나비촉진적PPARγ표체화지방세포적분화。결론：범소단백매계통통과HSP90조절PPARγ적표체，종이영향지방세포적분화。
AIM: To investigate the role of ubiquitin-proteasome system ( UPS) in adipocyte differentiation. METHODS:Differentiation of 3T3-L1 preadipocytes into adipocytes was induced by treatment with insulin, 3-isobutyl-1-methylxanthine and dexamethasone.Western blot and immunoprecipitation were performed to detect the protein abundances and association, respectively.Oil red O staining was used to determine the intracellular lipid of 3T3-L1 adipocytes.The levels of mRNA were measured by reverse transcription polymerase chain reaction ( RT-PCR) .RESULTS:UPS inhibitor bortezomib (BZM) suppressed the differentiation of 3T3-L1 pre-adipocytes, evidenced by reduced intracellular content of triglyceride, and decreased mRNA expression of adipogenic marker proteins such as adiponectin and adipocyte protein 2.In contrast, administration of sildenafil (SDN), an activator of protein kinase G which was also found to activate UPS, promo-ted adipocyte differentiation.In addition, BZM treatment decreased the expression of heat shock protein 90 (HSP90) and peroxisome proliferator-activated receptor gamma ( PPARγ) in the soluble fraction and reduced association of HSP90 and PPARγ.Furthermore, HSP90-specific N-terminal inhibitor geldanamic mitigated SDN-induced increase in PPARγlevel and 3T3-L1 cell differentiation.CONCLUSION:UPS modulates HSP90-dependent PPARγstability, thus leading to pro-motion of adipocyte differentiation.
