COX-2对棕榈酸或油酸诱导重新脂化HSC-T6 Acsl家族基因表达的影响

发布时间：2018-03-30 07:15

本文选题：COX-2　切入点：HSC-T6　出处：《南华大学》2014年硕士论文

【摘要】：目的：本章主要探讨棕榈酸或油酸对重新脂化HSC-T6的影响及COX-2在棕榈酸或油酸诱导的HSC-T6重新脂化过程中对各种Acsl表达影响。方法: 1、使用含有200mmol/L棕榈酸或20μg/ml油酸的DMEM培养基处理HSC-T6；油红O染色检测棕榈酸和油酸干预48小时后HSC-T6细胞内脂滴的变化，观察棕榈酸和油酸对HSC-T6细胞重新脂化的影响。 2、利用本实验构建的pYr-1.1-COX-2shRNA，将其转染至HSC-T6中，然后在荧光显微镜下观察并评估其转染效率。 3、Western Blot：COX-2shRNA干扰COX-2的表达后再分别给予棕榈酸或油酸处理细胞，Western blot检测各组HSC-T6细胞内COX-2，Acsl1，Acsl3，Acsl4，Acsl5，Acsl6的表达。结果： 1、200mmol/L棕榈酸或20μg/ml油酸干预48小时后均能够促进HSC-T6内脂滴的增多；棕榈酸干预组脂质沉积明显高于空白对照组，两者比较具有显著性差异（P 0.05）；油酸干预组脂质沉积明显高于空白对照组，两者比较具有显著性差异（P 0.05）。 2、转染48h后COX-2shRNA在荧光显微镜下观察转染效率达70%以上，符合本实验要求。 3、200mmol/L棕榈酸或20μg/ml油酸能够上调了HSC-T6中COX-2的表达；油酸干预组明显高于空白对照组，两者比较具有显著性差异（P 0.05）；棕榈酸干预组明显高于空白对照组，两者比较具有显著性差异（P 0.05）；与棕榈酸处理组或者油酸处理组比较，，COX-2shRNA+棕榈酸和COX-2shRNA+油酸明显降低了COX-2的表达，两者比较具有显著性差异（P 0.05）。 4、200mmol/L棕榈酸或20μg/ml油酸能够上调HSC-T6中Acsl1，Acsl5, Acsl6的表达，具有显著性差异（P 0.05）；棕榈酸处理组和油酸处理组均对Acsl3、Acsl4的表达无作用，统计无意义（P0.05）；与棕榈酸处理组和油酸处理组比较，COX-2shRNA+棕榈酸组和COX-2shRNA+油酸组进一步上调Acsl5、Acsl6的表达，具有显著性差异（P 0.05）；与棕榈酸组和油酸组比较，COX-2shRNA+棕榈酸组和COX-2shRNA+油酸组对Acsl1、Acsl3、Acsl4的表达无作用，统计无意义（P0.05）。结论： 1、棕榈酸或油酸均能够诱导HSC-T6重新脂化。 2、棕榈酸或油酸均能够上调HSC-T6中Acsl1、Acsl5、Acsl6的表达。 3、COX-2对棕榈酸或油酸上调Acsl5、Acsl6的表达具有拮抗作用。
[Abstract]:Objective:. In this chapter, the effects of palmitic acid or oleic acid on HSC-T6 and the effect of COX-2 on Acsl expression in palmitic or oleic acid-induced HSC-T6 resetting were studied. Methods:. 1. HSC-T6 was treated with DMEM medium containing 200mmol/L palmitic acid or 20 渭 g/ml oleic acid. The changes of lipid droplets in HSC-T6 cells were detected by oil red O staining after 48 hours of intervention by palmitic acid and oleic acid, and the effects of palmitic acid and oleic acid on the resaturation of HSC-T6 cells were observed. 2. PYr-1.1-COX-2shRNAs were used to transfect pYr-1.1-COX-2shRNAs into HSC-T6, and then the transfection efficiency was evaluated under fluorescence microscope. (3) Western Blot:COX-2shRNA interfered with the expression of COX-2 and then was treated with palmitic acid or oleic acid to detect the expression of HSC-T6 cells by Western blot. Results:. After 48 hours of 1200mmol/L palmitic acid or 20 渭 g/ml oleic acid treatment, lipid droplets in HSC-T6 were increased, and lipid deposition in palmitic acid intervention group was significantly higher than that in blank control group. The lipid deposition in the oleic acid intervention group was significantly higher than that in the blank control group, and there was a significant difference between the two groups (P 0.05). 2After 48 hours of transfection, the transfection efficiency of COX-2shRNA was more than 70% under fluorescence microscope, which met the requirements of this experiment. 3200mmol/L palmitic acid or 20 渭 g/ml oleic acid could up-regulate the expression of COX-2 in HSC-T6, oleic acid intervention group was significantly higher than blank control group, the difference was significant (P 0.05), palmitic acid intervention group was significantly higher than blank control group, Compared with palmitic acid group or oleic acid treatment group, the expression of COX-2 was significantly decreased in the two groups, and the difference was significant (P 0.05). 4200mmol/L palmitic acid or 20 渭 g/ml oleic acid could upregulate the expression of Acsl1, Acsl5, Acsl6 in HSC-T6, there was significant difference between palmitic acid group and oleic acid treatment group. Compared with palmitic acid treatment and oleic acid treatment, COX-2shRNA palmitic acid group and COX-2shRNA oleic acid group further upregulated the expression of Acsl5 + Acsl6. Compared with palmitic acid group and oleic acid group, COX-2shRNA palmitic acid group and COX-2shRNA oleic acid group had no effect on the expression of Acsl1, Acsl3, Acsl4, and had no statistical significance (P 0.05). Conclusion:. 1. Palmitic acid or oleic acid could induce HSC-T6 to be resollified. 2, both palmitic acid and oleic acid could up-regulate the expression of Acsl1, Acsl5 and Acsl6 in HSC-T6. 3 COX-2 antagonizes palmitic acid or oleic acid to up-regulate the expression of Acsl5 and Acsl6.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R575.2

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