COX-2对HSC-T6细胞中Acsl家族相关基因表达的影响

发布时间：2018-04-28 09:55

本文选题：COX-2 + shRNA　；参考：《南华大学》2014年硕士论文

【摘要】：目的研究COX-2对HSC-T6细胞中Acsl家族相关基因表达的影响，明确COX-2调节HSC-T6细胞脂类代谢的作用环节。方法 1、实验分组：HSC-T6细胞组（空白组），pYr-1.1-COX-2shRNA组（COX-2shRNA组）；pYr-1.1-NC组（空载体组）。 2、将构建好的pYr-1.1-COX-2shRNA及pYr-1.1-NC分别转染至HSC-T6细胞中，并于转染后48小时用荧光倒置显微镜观察转染效率。 3、利用Real-time PCR技术检测转染后各组细胞内COX-2、Acsl1、Acsl3、Acsl4、Acsl5、Acsl6基因表达情况。 4、根据Real-time PCR技术检测结果，选取Acsl家族中有意义的基因，再利用Western Blot技术检测HSC-T6细胞中Acsl家族相关基因的蛋白表达。结果 1、转染HSC-T6细胞48小时后，在荧光倒置显微镜下可见细胞质中分布着绿色荧光，且转染效率达75%以上。 2、HSC-T6细胞不同组别中COX-2mRNA表达情况：COX-2mRNA在COX-2shRNA组中表达低于空白组和空载体组，统计学处理有显著意义(p＜0.05)。 3、HSC-T6细胞不同组别中Acsl家族相关基因mRNA表达情况：COX-2shRNA组中Acsl6mRNA表达较空白组及空载体组明显增加，统计学处理有显著意义(p＜0.05)，，但Acsl1、Acsl3、Acsl4、Acsl5mRNA表达变化不大，没有统计学意义（p＞0.05）。 4、HSC-T6细胞不同组别中Acsl6蛋白质表达情况：COX-2shRNA组中Acsl6蛋白质表达较空白组及空载体组明显增加，统计学处理有显著意义(p＜0.05)。结论 1、靶向COX-2的shRNA可以明显沉默HSC-T6细胞中COX-2的表达。 2、在大鼠HSC-T6细胞中，Acsl1、Acsl3、Acsl4、Acsl5、Acsl6五种亚型均有表达；但沉默HSC-T6细胞COX-2表达后，仅Acsl6mRNA表达明显增强，Acsl1、Acsl3、Acsl4、Acsl5mRNA的表达无显著作用。 3、沉默HSC-T6细胞COX-2表达后，Acsl6蛋白质表达增加。 4、COX-2可能通过下调Acsl6的表达来调控HSC-T6细胞的脂类代谢。
[Abstract]:Purpose To study the effect of COX-2 on the expression of Acsl family related genes in HSC-T6 cells and to clarify the role of COX-2 in regulating lipid metabolism in HSC-T6 cells. Method 1. HSC-T6 cells were divided into two groups: pYr-1.1-COX-2shRNA group (pYr-1.1-COX-2shRNA group) and pYr-1.1-NC group (empty vector group). 2. The constructed pYr-1.1-COX-2shRNA and pYr-1.1-NC were transfected into HSC-T6 cells respectively, and the transfection efficiency was observed by fluorescence inverted microscope 48 hours after transfection. 3. Real-time PCR technique was used to detect the expression of Acsl6 gene in the transfected cells. 4According to the results of Real-time PCR technique, the significant genes of Acsl family were selected and the protein expression of Acsl family related genes in HSC-T6 cells was detected by Western Blot technique. Result 1. Green fluorescence was observed in the cytoplasm of HSC-T6 cells 48 hours after transfection, and the transfection efficiency was over 75%. 2The expression of COX-2mRNA in different groups of HSC-T6 cells was significantly lower in COX-2shRNA group than that in blank group and empty vector group (P < 0.05). 3The expression of Acsl family related gene mRNA in different groups of HSC-T6 cells was significantly higher in the group of Acsl family associated with mRNA than in the control group and empty vector group (P < 0.05), but there was no significant difference in the expression of Acsl6mRNA mRNA between the two groups (p > 0.05), but there was no significant difference in the expression of Acsl6mRNA gene between the two groups (p > 0.05). The expression of Acsl6mRNA family related gene in different groups of HSC-T6 cells was significantly higher than that in the control group and empty vector group. (4) the expression of Acsl6 protein in different groups of HSC-T6 cells was significantly higher than that in control group and empty vector group, and there was significant difference in Acsl6 protein expression between two groups (P < 0.05). Conclusion 1. ShRNA targeting COX-2 could significantly inhibit the expression of COX-2 in HSC-T6 cells. 2. All the five subtypes of Acsl1, Acsl3, Acsl4, Acsl5 and Acsl6 were expressed in rat HSC-T6 cells, but after silencing the expression of COX-2 in HSC-T6 cells, only the expression of Acsl6mRNA increased significantly. 3. After silencing the expression of COX-2 in HSC-T6 cells, the expression of Acsl6 protein increased. 4 COX-2 may regulate lipid metabolism of HSC-T6 cells by down-regulating the expression of 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】：R575.5

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