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赵梦鸽, 杨慧敏, 蒋翠花, 张健, 殷志琦. 青钱柳三萜化合物对游离脂肪酸诱导的脂肪变性的干预作用[J]. 中国药科大学学报, 2018, 49(3): 333-340. DOI: 10.11665/j.issn.1000-5048.20180312
引用本文: 赵梦鸽, 杨慧敏, 蒋翠花, 张健, 殷志琦. 青钱柳三萜化合物对游离脂肪酸诱导的脂肪变性的干预作用[J]. 中国药科大学学报, 2018, 49(3): 333-340. DOI: 10.11665/j.issn.1000-5048.20180312
ZHAO Mengge, YANG Huimin, JIANG Cuihua, ZHANG Jian, YIN Zhiqi. Intervention effects of the triterpenoids from Cyclocarya paliurus on free fatty acids-induced steatosis in HepG2 cells[J]. Journal of China Pharmaceutical University, 2018, 49(3): 333-340. DOI: 10.11665/j.issn.1000-5048.20180312
Citation: ZHAO Mengge, YANG Huimin, JIANG Cuihua, ZHANG Jian, YIN Zhiqi. Intervention effects of the triterpenoids from Cyclocarya paliurus on free fatty acids-induced steatosis in HepG2 cells[J]. Journal of China Pharmaceutical University, 2018, 49(3): 333-340. DOI: 10.11665/j.issn.1000-5048.20180312

青钱柳三萜化合物对游离脂肪酸诱导的脂肪变性的干预作用

Intervention effects of the triterpenoids from Cyclocarya paliurus on free fatty acids-induced steatosis in HepG2 cells

  • 摘要: 为探究青钱柳三萜化合物对非酒精性脂肪肝(NAFLD)的治疗作用,采用混合脂肪酸(FFAs)诱导HepG2细胞建立体外脂肪变性模型。MTT法筛选出无细胞毒性的青钱柳三萜化合物,通过检测其对胞内三酰甘油(TG)和超氧化物歧化酶(SOD)的作用筛选出具有良好降脂和抗氧化活性的青钱柳单体化合物 4 [2α,3α,23-三羟基-12,20(30)-二烯-28-乌苏酸,TUA]。通过油红O染色检测TUA对游离脂肪酸诱导的HepG2细胞中脂滴含量的影响,并同时检测其对TG、活性氧(ROS)、丙二醛(MDA)和SOD的作用。通过Western blot检测核因子相关因子2(Nrf2)、血红素氧合酶1(HO-1)和NAD(P)H醌氧化还原酶1(NQO-1)的蛋白表达情况。实验结果显示,TUA可降低FFAs诱导的HepG2细胞脂质堆积和TG的含量,减少ROS及脂质过氧化物MDA的含量,同时升高SOD的活性。此外,TUA还能增加抗氧化相关蛋白Nrf2、HO-1和NQO-1的表达。当同时加入TUA和Nrf2抑制剂鸦胆子苦醇干预后,HepG2细胞内TG、ROS和MDA含量增加,而SOD的活性和HO-1、NQO-1的表达下降。实验结果表明,TUA可能对NAFLD具有一定治疗作用,其机制可能与Nrf2的激活有关。

     

    Abstract: To investigate the therapeutic effects of triterpenoids from Cyclocarya paliurus on non-alcoholic fatty liver disease(NAFLD), the model of NAFLD in HepG2 cells was induced by free fatty acids(FFAs). Cytotoxicity of the triterpenoids from C. paliurus was determined by MTT method, and the effects of triterpenoids without cytotoxicity on intracellular triglyceride(TG)and superoxide dismutase(SOD)were detected by the kits. Data indicated that compound 4 [2α, 3α, 23-trihydroxy-12, 20(30)-dien-28-ursolic acid, TUA] had hypolipidemic and antioxidant activities. After being treated with TUA and FFAs for 24 h, the intracellular lipid content was observed using Oil Red O staining, and intracellular TG, malondialdehyde(MDA), SOD and reactive oxygen species(ROS)levels were determined by the assay kits. The protein expression of nuclear factor erythroid 2-related factor 2(Nrf2), heme oxygenase-1(HO-1)and NAD(P)H quinone oxidoreductase 1(NQO-1)were measured by Western blot. The results showed that TUA significantly increased SOD activity, and decreased intracellular TG, ROS and MDA levels in FFAs-induced HepG2 cells. Moreover, TUA dramatically improved Nrf2, NQO-1, and HO-1 expression. However, the dramatic increase in TG, ROS, MDA levels and the reduction in SOD, NQO-1 and HO-1 expression following Nrf2 inhibitor brusatol treatment were observed. In conclusion, these results suggest that TUA has the therapeutic effect on NAFLD which may be associated with Nrf2 activation.

     

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