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目的初步探讨整合素β1基因在RAW264.7细胞摄脂过程中的作用。方法针对整合素β1基因的不同靶点设计并化学合成3条具有阳性转染率的siRNA,用脂质体法转染RAW264.7细胞,并根据转染的方式将实验分为空白对照组、转染siRNA阴性对照组、脂质体对照组和转染筛选出的阳性转染率最高的siRNA组。用Cy3标记siRNA荧光显微镜下检测转染效率,Real-time PCR筛选沉默效率最高的siRNA,细胞黏附性实验检测细胞黏附性的改变,脂质油红O化学染色法检测细胞的脂质摄取量的变化,上述检测均重复测定3次。结果成功将siRNA转染入RAW264.7细胞,并成功筛选出沉默效率最高(65%)的siRNA为siRNA1,转染后实验组整合素β1mRNA表达明显低于其他组,细胞的黏附性从其他组的70%以上降低到53%,泡沫细胞转化率由其他组的90%以上显著降低到17%,摄脂能力显著降低,与其他组比较,差异有统计学意义(P<0.05)。结论整合素β1基因明显影响RAW264.7细胞的摄脂功能,进而影响RAW264.7细胞向泡沫细胞的转化。
Objective To investigate the role of integrin β1 in liposuction in RAW264.7 cells. Methods Three siRNAs with positive transfection efficiency were designed and synthesized based on the different targets of integrin β1 gene. Lipofectamine 2000 was transfected into RAW264.7 cells. According to the transfection method, the experiment was divided into blank control group, The siRNA negative control group, the liposome control group and the transfected screened siRNA group with the highest positive transfection rate were transfected. Transfection efficiency was detected by Cy3-labeled siRNA fluorescence microscope, siRNA with the highest silencing efficiency was screened by Real-time PCR, cell adhesion was detected by cell adhesion assay, and lipid uptake by lipid-oil red O staining Changes, the above tests were repeated 3 times. Results The siRNA was successfully transfected into RAW264.7 cells and the siRNA with the highest silencing efficiency (65%) was successfully screened out. The expression of integrin β1 mRNA in the experimental group was significantly lower than that in other groups after transfection. (P <0.05). Compared with the other groups, the foam cell transformation rate decreased significantly from above 90% to 17% and the lipogenic capacity decreased significantly from 70% to 53%. Conclusion The integrin β1 gene obviously affects the lipofunction of RAW264.7 cells, and then affects the transformation of RAW264.7 cells into foam cells.