清肝降脂方对脂肪变性LO2细胞HO-1、Egr-1基因及蛋白表达的影响

发布时间：2018-11-25 09:28

【摘要】：目的:通过建立游离脂肪酸体外诱导的LO2细胞非酒精脂肪肝模型,探讨不同剂量的清肝降脂方对肝脏脂肪变LO2细胞HO-1、Egr-1基因及蛋白表达水平的影响。方法:建立游离脂肪酸诱导脂肪变性LO2肝细胞体外模型。人肝LO2细胞采用含10%胎牛血清的RPMI 1640培养基培养,待细胞均匀分布,随机分为6组:正常对照组、模型组、清肝降脂方高剂量组、中剂量组、低剂量组、水林佳对照组,每组6瓶。除正常对照组外,其余5组细胞均以0.5 mmol/L的游离脂肪酸诱导LO2非酒精脂肪肝细胞模型,诱导24 h后,各组随机抽取1瓶进行油红O染色,观察LO2细胞的脂肪化程度。确定模型诱导成功后,正常对照组和模型组分别加入10%正常大鼠血清,低、中、高剂量清肝降脂方药物血清组及水林佳药物血清组添加10%含相对应药物的大鼠血清,培养24h后,收集细胞;采用RT-PCR及Western blot法检测血红素氧化酶-1(HO-1)、早期生长反应因子-1(Egr-1)的基因及蛋白表达水平。结果:与正常对照组比较,模型对照组LO2细胞HO-1、Egr-1 mRNA和蛋白表达有所增加;与模型对照组比较,自拟清肝降脂方三剂量组和对照药物组HO-1 mRNA及蛋白表达有所增高,Egr-1mRNA和蛋白表达明显降低;与对照药物组比较,清肝降脂方高剂量组HO-1 mRNA表达有所增高,Egr-1 mRNA表达有所降低。结论:Egr-1高表达可能会导致肝损伤机制之一;HO-1高表达可能是保护肝损伤机制之一;两者可能都是通过调节TNF-α表达来调节NAFLD细胞的损伤进展。清肝降脂方可明显改善LO2细胞Egr-1、HO-1 mRNA及蛋白的表达,可能为其治疗机制、临床诊断和判断进展参考指标。
[Abstract]:Aim: to study the effect of Qinggan Jiangzhi recipe (QQQQF) on the expression of HO-1,Egr-1 gene and protein in LO2 cells induced by free fatty acids in vitro. Methods: the model of LO2 hepatocytes induced by free fatty acids was established in vitro. Human liver LO2 cells were cultured on RPMI 1640 medium containing 10% fetal bovine serum and were randomly divided into 6 groups: normal control group, model group, high dose group, middle dose group, low dose group and Shuilingjia control group. Six bottles per group. In addition to the normal control group, the other five groups were treated with free fatty acids for 0.5 mmol/L to induce LO2 non-alcoholic fatty liver cells. After 24 h of induction, 1 bottle was randomly selected for oil red O staining in each group to observe the fatty degree of LO2 cells. After the model was induced successfully, the normal control group and the model group were added with 10% normal rat serum, the low, middle and high doses of Qinggan Jiangzhi prescription drug serum group and the water linjia drug serum group supplemented with 10% rat serum containing corresponding drugs, respectively. After 24 hours of culture, the cells were collected. The gene and protein expression of heme oxygenase-1 (HO-1) and early growth response factor-1 (Egr-1) were detected by RT-PCR and Western blot. Results: compared with the normal control group, the expression of HO-1,Egr-1 mRNA and protein in LO2 cells in the model control group was increased. Compared with the model control group, the expression of HO-1 mRNA and protein was increased and the expression of Egr-1mRNA and protein was significantly decreased in the three dose group and the control group. Compared with the control group, the expression of HO-1 mRNA was increased and the expression of Egr-1 mRNA was decreased in the high dose group of Qinggan Jiangzhi recipe. Conclusion: the high expression of Egr-1 may lead to one of the mechanisms of liver injury, the high expression of HO-1 may be one of the mechanisms of protecting liver injury, and both of them may regulate the progression of NAFLD cell injury by regulating the expression of TNF- 伪. Qinggan Jiangzhi decoction can obviously improve the expression of Egr-1,HO-1 mRNA and protein in LO2 cells, which may be a reference index for its therapeutic mechanism, clinical diagnosis and judgement of progress.
【作者单位】：辽宁中医药大学;辽宁中医药大学附属医院;
【基金】：沈阳市科技创新专项基金-生物与制药科技攻关专项项目(F13-208-9-00)
【分类号】：R285

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