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纸质出版日期:2017
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许光远, 孙文, 郭璇, 等. 青钱柳总皂苷对游离脂肪酸诱导的H4-ⅡE细胞脂肪代谢的影响及作用机制[J]. 中国实验方剂学杂志, 2017,23(15):124-129.
XU Guang-yuan, SUN Wen, GUO Xuan, et al. Effect and Mechanism of CPS on Fat Metabolism in H4-ⅡE Cells Induced by Free Fatty Acids[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(15): 124-129.
许光远, 孙文, 郭璇, 等. 青钱柳总皂苷对游离脂肪酸诱导的H4-ⅡE细胞脂肪代谢的影响及作用机制[J]. 中国实验方剂学杂志, 2017,23(15):124-129. DOI: 10.13422/j.cnki.syfjx.2017150124.
XU Guang-yuan, SUN Wen, GUO Xuan, et al. Effect and Mechanism of CPS on Fat Metabolism in H4-ⅡE Cells Induced by Free Fatty Acids[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(15): 124-129. DOI: 10.13422/j.cnki.syfjx.2017150124.
目的:研究青钱柳总皂苷(CPS)对游离脂肪酸(FFA)诱导的H4-ⅡE肝细胞脂肪代谢的影响和作用机制。方法:以FFA诱导的H4-ⅡE大鼠肝细胞为模型,培养72 h后,采用Neutral red法检测CPS(0,0.05,0.10,0.20,0.40 g·L-1)对细胞活性的影响;根据细胞活性检测结果选择CPS低剂量组(CPS-L,0.10 g·L-1)和高剂量组(CPS-H,0.20 g·L-1)干预,油红O(Oile red)染色并测定细胞内脂质含量;药物干预2 h后实时荧光定量PCR(Real-time PCR)检测肝细胞相应基因表达,药物干预30 min后,利用蛋白免疫印迹法(Western blot)检测细胞相应蛋白磷酸化水平。结果:与正常组比较,0.10,0.20 g·L-1 CPS干预后肝细胞活性明显升高(P< 0.05);干预72 h后,Oile red染色显示FFA(20 μmol·L-1)诱导后呈现大片红染的脂滴,细胞脂质含量显著增加(P< 0.01),CPS干预后细胞中红染的脂滴减少,细胞脂质含量较FFA(20 μmol·L-1)诱导组明显减少(P< 0.05,P< 0.01);与模型组比较,CPS-L和CPS-H组腺苷酸活化蛋白激酶(AMPK),乙酰CoA羧化酶(ACC)蛋白磷酸化水平明显升高(P< 0.05,P< 0.01);胆固醇应答元件结合蛋白1c(SREBP1c),脂肪酸合成酶(FAS)mRNA表达水平明显降低(P< 0.05,P< 0.01)。结论:CPS能够显著降低体外FFA诱导肝细胞脂质沉积,可能是通过上调AMPK,ACC蛋白磷酸化水平,下调SREBP1c,FAS mRNA表达而发挥作用。
Objective: To explore the effect and mechanism of Cyclocarya paliurus saponin (CPS) on FFA-induced H4-ⅡE in hepatic cells. Method: FFA-induced H4-ⅡE liver cells of rat were cultivated for 72 h
and then exposed to various concentrations of CPS (0
0.05
0.10
0.20
0.40 g·L-1)
and their viability was measured by Neutral red. According to the results of CPS cell activity
these cells were treated in the low-dose group (CPS-L
0.10 g·L-1) and the high-dose group (CPS-H
0.20 g·L-1)
Oile red staining was performed to determine lipid content in the cells. After treatment of the cells for 2 h
mRNA levels were measured by Real-time PCR
while phosphorylation levels of target proteins were detected by Western blot after treatment for 30 min. Result: It was found that the cell viability indexes in the cells intervened by 0.10
0.20 g·L-1 CPS increased significantly
compared with normal group (P<0.05). After treatment for 72 h
Oile red staining showed that FFA (20 μmol·L-1)-induced cells contained a large number of red lipid droplets
and lipid content increased significantly (P<0.01). However
after treatment CPS
red lipid droplets decreased
and lipid content significantly decreased (P<0.05
P<0.01). After intervention with CPS
phosphorylation levels of mitogen-activated protein kinase (AMPK)
and acetylcoacarboxylase (ACC) in CPS-L and CPS-H groups were higher than those in control group (P<0.05
P<0.01). And sterol regulatory element-binding protein 1c (SREBP1c) and fatty acid synthase (FAS) mRNA expressions in CPS-L and CPS-H groups increased
compared with control group (P<0.05
P<0.01). Conclusion: CPS can play a significant role in reducing FFA-induced lipid deposition in liver cells in vitro; The mechanism may be correlated with up-regulation of AMPK
ACC protein and phosphorylation levels
and down-regulation of SREBP1c
FAS mRNA expressions.
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