本文選題：COX-2　切入點：HSC-T6　出處：《南華大學》2014年碩士論文

【摘要】：目的： 本章主要探討棕櫚酸或油酸對重新脂化HSC-T6的影響及COX-2在棕櫚酸或油酸誘導的HSC-T6重新脂化過程中對各種Acsl表達影響。 方法: 1、使用含有200mmol/L棕櫚酸或20μg/ml油酸的DMEM培養(yǎng)基處理HSC-T6；油紅O染色檢測棕櫚酸和油酸干預48小時后HSC-T6細胞內脂滴的變化，觀察棕櫚酸和油酸對HSC-T6細胞重新脂化的影響。 2、利用本實驗構建的pYr-1.1-COX-2shRNA，將其轉染至HSC-T6中，然后在熒光顯微鏡下觀察并評估其轉染效率。 3、Western Blot：COX-2shRNA干擾COX-2的表達后再分別給予棕櫚酸或油酸處理細胞，Western blot檢測各組HSC-T6細胞內COX-2，Acsl1，Acsl3，Acsl4，Acsl5，Acsl6的表達。 結果： 1、200mmol/L棕櫚酸或20μg/ml油酸干預48小時后均能夠促進HSC-T6內脂滴的增多；棕櫚酸干預組脂質沉積明顯高于空白對照組，兩者比較具有顯著性差異（P 0.05）；油酸干預組脂質沉積明顯高于空白對照組，兩者比較具有顯著性差異（P 0.05）。 2、轉染48h后COX-2shRNA在熒光顯微鏡下觀察轉染效率達70%以上，符合本實驗要求。 3、200mmol/L棕櫚酸或20μg/ml油酸能夠上調了HSC-T6中COX-2的表達；油酸干預組明顯高于空白對照組，兩者比較具有顯著性差異（P 0.05）；棕櫚酸干預組明顯高于空白對照組，兩者比較具有顯著性差異（P 0.05）；與棕櫚酸處理組或者油酸處理組比較，，COX-2shRNA+棕櫚酸和COX-2shRNA+油酸明顯降低了COX-2的表達，兩者比較具有顯著性差異（P 0.05）。 4、200mmol/L棕櫚酸或20μg/ml油酸能夠上調HSC-T6中Acsl1，Acsl5, Acsl6的表達，具有顯著性差異（P 0.05）；棕櫚酸處理組和油酸處理組均對Acsl3、Acsl4的表達無作用，統(tǒng)計無意義（P0.05）；與棕櫚酸處理組和油酸處理組比較，COX-2shRNA+棕櫚酸組和COX-2shRNA+油酸組進一步上調Acsl5、Acsl6的表達，具有顯著性差異（P 0.05）；與棕櫚酸組和油酸組比較，COX-2shRNA+棕櫚酸組和COX-2shRNA+油酸組對Acsl1、Acsl3、Acsl4的表達無作用，統(tǒng)計無意義（P0.05）。 結論： 1、棕櫚酸或油酸均能夠誘導HSC-T6重新脂化。 2、棕櫚酸或油酸均能夠上調HSC-T6中Acsl1、Acsl5、Acsl6的表達。 3、COX-2對棕櫚酸或油酸上調Acsl5、Acsl6的表達具有拮抗作用。
[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.
【學位授予單位】：南華大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R575.2

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