Rac1-siRNA抑制大鼠视网膜NF-κB表达的实验研究

Experimental study of small interfering RNA targeting Rac1 inhibits NF-κB expression in retina of rat model

目的观察Rac1-siRNA重组载体对大鼠视网膜核因子-κB(nuclearfactor-κB,NF-κB)表达的抑制作用。方法将25只成年SD大鼠,采用光动力法诱导双眼视网膜静脉阻塞后,一眼玻璃体内转染Rac1-siRNA重组载体作为基因干预组,另一眼注射空白载体作为空白对照组。同时选取正常同龄SD大鼠25只一眼玻璃体内转染Rac1-siRNA重组载体作为空白干预组,另一眼作为阴性对照组。免疫组织化学法和RT-PCR法检测各组NF-κB的表达情况。结果NF-κB免疫组织化学染色后,空白干预组、空白对照组、基因干预组OD值分别为:28.698±4.064、93.462±12.398、64.799±10.895,组间比较差异均有统计学意义(P<0.05)。RT-PCR结果显示:在基因干预组和空白对照组中与空白干预组相比表达较强,空白对照组最强。各组NF-κB表达率差异具有统计学意义(P<0.05)。结论Rac1-siRNA重组载体能有效抑制视网膜内NF-κB的表达。

Objective To observe the inhibitory effects of small interfering RNA targeting Racl （Racl-siRNA） on nuclear factor-κB（NF-κB） expression in retina of rat model. Methods Twenty-five SD rats＇ retinal vein were laser photocoagulated,and injected the Racl-siRNA vector into vltrous as the group of gene intervention, and injected empty vector in another eye as blank contrast group;Racl-siRNA vector was injected in one of the eyes in other 25 SD normal rats as blank intervention group. Two weeks after trangeetion, the expression of NF-κB was detected by immunohistochemistry and RT-PCR. Results The OD of in the groups of blank intervention,blank contrast, gene intervention were 28. 698 ± 4. 064,93. 462 ± 12.398, and 64.799 ± 10. 895, respectively. There were significant differences between each two groups （ P 〈 0. 05 ）. The results indicated that in the group of gene intervention and blank contrast group, the expression of NF-κB of RT-PCR is higher than blank intervention group, and the blank contrast group was the highest. There was signiflcant difference （ P 〈 0. 05）. Conclusion Reconstructive vector Racl-siRNA can effectively inhabit the expression of NF-κB in retina.