CAJ | 학술논문

目的：研究天麻素( GSTD)对油酸( OA)诱导的 HL-7702细胞脂肪蓄积的抑制作用,并探讨其可能的细胞信号通路。方法以噻唑蓝( MTT)法测定GSTD对HL-7702细胞存活率的影响；以1 mmol·L-1的OA处理24 h诱导细胞脂肪变性,同时加入不同浓度的GSTD,油红O( ORO)染色观察细胞脂肪蓄积情况并测定细胞内三酰甘油( TG)含量；以Western blot检测GSTD处理后细胞中AMPKα和ACC的磷酸化水平；以compound C处理细胞,研究其对GSTD药效的影响。结果 GSTD浓度≤3386．5μmol·L-1时对HL-7702细胞没有明显毒性。 OA处理24 h后细胞中出现大量脂滴蓄积,TG含量明显增加,但同时加入浓度为169．3或338．7μmol·L-1的GSTD可明显抑制HL-7702细胞的脂肪蓄积,并使TG含量分别平均下降35%和43．6%(与单加OA的细胞比较P<0．01)。 GSTD处理后能时间和浓度依赖性地增加细胞中的p-AMPKα和p-ACC水平,compound C能完全阻断GSTD对AMPK通路的激活作用以及减少肝细胞TG蓄积的作用。结论 GSTD可明显抑制OA诱导的HL-7702细胞脂肪蓄积,降低TG含量,其作用依赖于细胞中AMPK通路的激活。
목적：연구천마소( GSTD)대유산( OA)유도적 HL-7702세포지방축적적억제작용,병탐토기가능적세포신호통로。방법이새서람( MTT)법측정GSTD대HL-7702세포존활솔적영향；이1 mmol·L-1적OA처리24 h유도세포지방변성,동시가입불동농도적GSTD,유홍O( ORO)염색관찰세포지방축적정황병측정세포내삼선감유( TG)함량；이Western blot검측GSTD처리후세포중AMPKα화ACC적린산화수평；이compound C처리세포,연구기대GSTD약효적영향。결과 GSTD농도≤3386．5μmol·L-1시대HL-7702세포몰유명현독성。 OA처리24 h후세포중출현대량지적축적,TG함량명현증가,단동시가입농도위169．3혹338．7μmol·L-1적GSTD가명현억제HL-7702세포적지방축적,병사TG함량분별평균하강35%화43．6%(여단가OA적세포비교P<0．01)。 GSTD처리후능시간화농도의뢰성지증가세포중적p-AMPKα화p-ACC수평,compound C능완전조단GSTD대AMPK통로적격활작용이급감소간세포TG축적적작용。결론 GSTD가명현억제OA유도적HL-7702세포지방축적,강저TG함량,기작용의뢰우세포중AMPK통로적격활。
Aim To study the inhibitory effect of gast-rodin (GSTD) on oleic acid (OA)-induced fat accu-mulation in HL-7702 cells and explore possible cellular signaling pathways. Methods The MTT method was used to study the impact of GSTD on cell viability in HL-7702 cells. Cellular steatosis was induced by 1 mmol·L-1 of OA administration for 24 h, and differ-ent concentrations of GSTD were added at the same time. Oil red O ( ORO) staining was used to determine fat accumulation in cells, and intracellular triglyceride ( TG) contents were assayed. Western blot was used to determine the phosphorylation levels of AMPKα and ACC in cells after GSTD administration. Compound C was used to treat the cells in order to study its influ-ence on the efficacies of GSTD. Results GSTD had no obvious toxicity in HL-7702 cells when its concen-tration was≤3 386. 5 μmol · L-1 . After 24 h of OA administration, there were large amounts of lipid drop-lets accumulated in HL-7702 cells, and intracellular TG contents greatly increased as well. However, when 169. 3 or 338. 7 μmol · L-1 of GSTD was added to-gether with OA, fat accumulation in cells was greatly inhibited, and intracellular TG contents were reduced averagely by 35% and 43 . 6%, respectively ( P<0. 01 vs OA alone ) . After administration, GSTD could in-crease the levels of p-AMPKα and p-ACC in HL-7702 cells time and dose dependently. Compound C could completely abolish the stimulating activity of GSTD on AMPK pathway and block its reducing effect on hepatic TG accumulation. Conclusions GSTD greatly inhibits OA-induced fat accumulation and reduces intracellular TG contents in HL-7702 cells;the efficacy of GSTD is dependent on the activation of cellular AMPK pathway.