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[干扰HepG2细胞中肝细胞核因子-1α对载脂蛋白M、载脂蛋白A-I及胆固醇代谢相关关键酶表达的影响]

[Effect of interfering hepatocyte nuclear factor-1 alfa in HepG2 on the expressions of apoM, apoA-I and the correlative key enzyme of cholesterol metabolism].

作者信息

Zhang Yao, Chen Chang-Jie, Yang Qing-Ling, Cheng Long-Qiang, Wang Hui, Huang Li-Zhu

机构信息

Department of Biochemistry and Molecular Biology, Wannan Medical College, Wuhu Anhui 241000, China.

出版信息

Zhonghua Gan Zang Bing Za Zhi. 2011 Feb;19(2):121-6. doi: 10.3760/cma.j.issn.1007-3418.2011.02.012.

Abstract

To determine wether there were connections among hepatocyte nuclear factor-1 alfa (HNF-1a), liver receptor homolog-1 (LRH-1), apolipoprotein M (apoM) and to investigate the effects of HNF-1a in HepG2 on the expressions of apoM, apolipoprotein A-I (apoA-I) and the key enzymes in cholesterol metabolism and biotransformation. The mRNA expressions of apoM, LRH-1 and HNF-1a were detected by RT-PCR. HNF-1a was interfered and RT-PCR was used to detect the changes of apo M, apo A-I, Cyp7A1, farnesoid X receptor (FXR) and small heterodimer partner-1 (SHP-1). Western blot was used to detect the change of apo M protein. The expressions of apoM, LRH-1 and HNF-1amRNA were obviously higher in HCC tissue than that in para-cancer tissue (the vaule of t is -7.167, -7.075, -8.803, P less than 0.01 respectively). HNF-1a and LRH-1 positively correlated with the expression of apoM (r=0.353, P less than 0.01; r=0.523, P less than 0.01 respectively); RT-PCR and western blot results showed that the expressions of apoM, FXR and SHP-1 mRNA, could be obviously suppressed by HNF-1a interfering as compared to the negative controls by 47.4%, 47.9%and 65.2% (P less than 0.01) respectively, and the expression of apoM protein also decreased by 54.3% (F = 43.482, P less than 0.01). The expressions of HMGCR and CYP7A mRNA increased by 101.1% and 138.5% (P less than 0.01) respectively as compared to the negative control. But there is no effect on expression of apoA-I mRNA (F = 0.170, P more than 0.05). HNF-1a could promote cholesterol biotransformation by increasing the expression of apoM and the key enzymes in cholesterol metabolism and decreasing inhibiting factor. So HNF-1a provided protection against cardiovascular disease.

摘要

确定肝细胞核因子-1α(HNF-1α)、肝脏受体同源物-1(LRH-1)、载脂蛋白M(apoM)之间是否存在联系,并研究HNF-1α在HepG2细胞中对apoM、载脂蛋白A-I(apoA-I)表达以及胆固醇代谢和生物转化关键酶的影响。采用逆转录聚合酶链反应(RT-PCR)检测apoM、LRH-1和HNF-1α的mRNA表达。干扰HNF-1α后,用RT-PCR检测apo M、apo A-I、细胞色素P450 7A1(Cyp7A1)、法尼醇X受体(FXR)和小异二聚体伴侣-1(SHP-1)的变化。用蛋白质免疫印迹法(Western blot)检测apo M蛋白的变化。肝癌组织中apoM、LRH-1和HNF-1α mRNA的表达明显高于癌旁组织(t值分别为-7.167、-7.075、-8.803,P均小于0.01)。HNF-1α和LRH-1与apoM的表达呈正相关(r分别为0.353,P小于0.01;r为0.523,P小于0.01);RT-PCR和Western blot结果显示,与阴性对照相比,干扰HNF-1α后apoM、FXR和SHP-1 mRNA的表达分别明显受到抑制,抑制率分别为47.4%、47.9%和65.2%(P小于0.01),apoM蛋白表达也下降了54.3%(F = 43.482,P小于0.01)。与阴性对照相比,3-羟基-3-甲基戊二酰辅酶A还原酶(HMGCR)和CYP7A mRNA的表达分别增加了101.1%和138.5%(P小于0.01)。但对apoA-I mRNA的表达无影响(F = 0.170,P大于0.05)。HNF-1α可通过增加apoM和胆固醇代谢关键酶的表达以及减少抑制因子来促进胆固醇生物转化。因此,HNF-1α对心血管疾病具有保护作用。

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