CAJ | 학술논문

目的观察不同浓度二甲双胍对非酒精性脂肪肝细胞模型过氧化物酶体增殖物激活受体γ( PPARγ)共激活因子-1α( PGC-1α)基因表达的影响,以及PGC-1α的表达与肝脏细胞脂肪堆积的关系。方法用20μg/ml油酸(油酸以0.5%DMSO溶解)诱导L-02细胞72 h,形成非酒精性脂肪变性肝细胞模型。对照组加入含10%胎牛血清的普通1640培养基。在模型组中分别添加含2.5、5、7.5 mmol/L终浓度的二甲双胍培养基继续培养24 h 后收集细胞。采用 RT-PCR法检测L-02细胞PGC-1αmRNA的表达,采用三酰甘油( TG)酶法测定试剂盒(组织细胞)检测L-02细胞中TG的变化。结果当二甲双胍终浓度为7.5 mmol/L时,细胞内TG明显减少,与模型组比较差异有统计学意义( P<0.05)；二甲双胍终浓度为7.5 mmol/L组与终浓度为2.5 mmol/L组比较,细胞内TG明显减少,而细胞内PGC-1α mRNA表达量明显增多,差异有统计学意义( P<0.05)。与模型组比较,当二甲双胍终浓度为2.5、7.5 mmol/L时L-02细胞中PGC-1α mRNA表达量明显增多,差异有统计学意义( P<0.05)。L-02细胞中PGC-1α mRNA的表达与 TG 水平呈负相关性(r=-0.581,P<0.05)。随着二甲双胍浓度的增加,模型组L-02细胞线粒体损伤有所改善。结论非酒精性脂肪肝细胞模型中PGC-1α的表达减少而二甲双胍可改善PGC-1α的表达水平,PGC-1α的表达和细胞内TG的水平呈负相关,由此得出增加PGC-1α的表达水平可减少脂肪在肝脏的堆积。
목적관찰불동농도이갑쌍고대비주정성지방간세포모형과양화물매체증식물격활수체γ( PPARγ)공격활인자-1α( PGC-1α)기인표체적영향,이급PGC-1α적표체여간장세포지방퇴적적관계。방법용20μg/ml유산(유산이0.5%DMSO용해)유도L-02세포72 h,형성비주정성지방변성간세포모형。대조조가입함10%태우혈청적보통1640배양기。재모형조중분별첨가함2.5、5、7.5 mmol/L종농도적이갑쌍고배양기계속배양24 h 후수집세포。채용 RT-PCR법검측L-02세포PGC-1αmRNA적표체,채용삼선감유( TG)매법측정시제합(조직세포)검측L-02세포중TG적변화。결과당이갑쌍고종농도위7.5 mmol/L시,세포내TG명현감소,여모형조비교차이유통계학의의( P<0.05)；이갑쌍고종농도위7.5 mmol/L조여종농도위2.5 mmol/L조비교,세포내TG명현감소,이세포내PGC-1α mRNA표체량명현증다,차이유통계학의의( P<0.05)。여모형조비교,당이갑쌍고종농도위2.5、7.5 mmol/L시L-02세포중PGC-1α mRNA표체량명현증다,차이유통계학의의( P<0.05)。L-02세포중PGC-1α mRNA적표체여 TG 수평정부상관성(r=-0.581,P<0.05)。수착이갑쌍고농도적증가,모형조L-02세포선립체손상유소개선。결론비주정성지방간세포모형중PGC-1α적표체감소이이갑쌍고가개선PGC-1α적표체수평,PGC-1α적표체화세포내TG적수평정부상관,유차득출증가PGC-1α적표체수평가감소지방재간장적퇴적。
Objective To observe the different concentrations of metformin on the peroxisome proliferator-activated receptorγ( PPARγ) coactivator-1α ( PGC-1α) gene expression, and the relationship between lipide accumulation in nonalcoholic fatty liver cell model. Methods L-02 cells were treated by 20μg/ml oleic acid ( oleic acid was so-lutioned by 0.5% DMSO) for 72 h to induce the nonalcoholic fatty liver cell model. The control group added ordi-nary 1640 culture medium containing 10% fetal bovine serum. The model group cells were cultured in the medium containing 2.5, 5, 7.5 mmol/L concentrations of metformin and continue to cultivate 24 h then collected cells. Use RT-PCR analysis of PGC-1α mRNA expression, and triglycerides ( tissue) enzymatic assay kit to detect chan-ges of triglycerides in L-02 cells. Results When the concentration of metformin was 7. 5 mmol/L in L-02 cells the triglyceride levels were reduced significantly compared with the model group, the difference was statistically signifi-cant ( P<0.05 ) . When the concentration of metformin was 7.5 mmol/L the triglyceride levels was reduced signifi-cantly compared with the group containing 2. 5 mmol/L metformin,but the expression of PGC-1α mRNA was in-creased significantly. These differences were all statistically significant ( P <0.05 ) . When the concentration of metformin was 2.5, 5, 7.5 mmol/L with L-02 cells, the expression of PGC-1α mRNA was increased obviously compared with the model group and the difference was statistically significant ( P<0.05 ) . The expression of PGC-1α mRNA and the triglyceride levels showed a negative correlation with L-02 cells(r= -0.581,P<0.05). With the increasing of the concentration of metformin in L-02 cells, mitochondria injury was improved in the model group. Conclusion The expression of PGC-1αis reduced in nonalcoholic fatty liver cell model and metformin can improve the expression of PGC-1α. It shows a negative correlation between the expression of PGC-1α mRNA and the triglyceride levels, thus the increased expression of PGC-1α can reduce the accumulation of fat in the liver.