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目的体外观察熊果酸对肝星状细胞内活性氧(ROS)的产生、细胞凋亡、NF-κB核易位及X-连锁凋亡抑制蛋白(XIAP)mRNA表达的影响,探讨熊果酸诱导肝星状细胞凋亡的机制。方法取对数生长期的肝星状细胞(HSC-T6)进行分组,A组:瘦素(100 ng/mL)刺激组;B组:瘦素+熊果酸(50μmol/L)组;C组:瘦素+N-乙酰-L半胱氨酸(10 mmol/L)组;D组:空白对照组。检测DCF荧光强度(反映ROS水平)、细胞凋亡率、NF-κB(P65)核移位率、XIAP mRNA的表达。结果 (1)A组HSC-T6细胞内DCF荧光强度明显强于D组,B、C组细胞内ROS水平明显低于A组(均P<0.001)。(2)A组在48 h的HSC-T6细胞凋亡率低于D组(P<0.05);B组在48 h的HSC-T6凋亡率不仅明显高于A组(P<0.001),而且高于C、D组(均P<0.05)。(3)A组细胞NF-κB核移位率显著高于D组(P<0.001);B、C组的NF-κB核移位率均显著低于A组(均P<0.001)。(4)瘦素作用HSC-T6细24 h的XIAP mRNA表达显著高于D组(P<0.01);B、C组在12、24 h的表达均低于A组(P<0.01或P<0.05)。结论熊果酸能抑制瘦素刺激HSC-T6细胞引起的ROS产生,并能诱导HSC-T6细胞凋亡,其机制可能与抑制ROS对NF-κB的活化,下调XIAP基因表达有关。
Objective To observe the effect of ursolic acid on the production of reactive oxygen species (ROS), apoptosis, nuclear translocation of NF-κB and the expression of X-linked inhibitor of apoptosis protein (XIAP) in hepatic stellate cells in vitro. Induction of apoptosis in hepatic stellate cells. Methods HSC-T6 cells in logarithmic growth phase were divided into group A: leptin (100 ng / mL) stimulation group; group B: leptin + ursolic acid (50 μmol / L) group; C Group: Leptin + N-acetyl-L-cysteine (10 mmol / L) group; Group D: blank control group. DCF fluorescence intensity (ROS level), apoptosis rate, nuclear translocation rate of NF-κB (P65) and XIAP mRNA expression were detected. Results (1) Fluorescence intensity of DCF in HSC-T6 cells in group A was significantly higher than that in group D. ROS levels in cells in group B and C were significantly lower than those in group A (all P <0.001). (2) The apoptosis rate of HSC-T6 cells in group A at 48 h was lower than that in group D (P <0.05). The apoptosis rate of HSC-T6 at 48 h in group B was significantly higher than that in group A (P <0.001) But higher than those in group C and D (all P <0.05). (3) The nuclear translocation rate of NF-κB in group A was significantly higher than that in group D (P <0.001). The nuclear translocation rates of NF-κB in group B and C were significantly lower than those in group A (all P <0.001). (4) The expression of XIAP mRNA in HSC-T6 treated with leptin for 24 h was significantly higher than that in group D (P <0.01). The expression of XIAP in group B and C at 12 and 24 h was lower than that of group A (P <0.01 or P < 0.05). Conclusion UA can inhibit the production of ROS induced by leptin in HSC-T6 cells and induce the apoptosis of HSC-T6 cells. The mechanism may be related to the inhibition of ROS activation of NF-κB and the down-regulation of XIAP gene expression.