[1]陈新,成名翔,龚建平,等.沉默供体肝脏内核糖体蛋白S3基因减轻大鼠移植肝脏缺血再灌注损伤[J].第三军医大学学报,2011,33(14):1441-1445.
 Chen Xin,Cheng Mingxiang,Gong Jianping,et al.Silencing ribosome protein S3 gene in donor livers reduces ischemia/reperfusion injury following orthotopic liver transplantation in rats[J].J Third Mil Med Univ,2011,33(14):1441-1445.
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沉默供体肝脏内核糖体蛋白S3基因减轻大鼠移植肝脏缺血再灌注损伤(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
33卷
期数:
2011年第14期
页码:
1441-1445
栏目:
论著
出版日期:
2011-07-30

文章信息/Info

Title:
Silencing ribosome protein S3 gene in donor livers reduces ischemia/reperfusion injury following orthotopic liver transplantation in rats
作者:
陈新成名翔龚建平陈勇涂兵
重庆医科大学附属第二医院肝胆外科
Author(s):
Chen Xin Cheng Mingxiang Gong Jianping Chen Yong Tu Bing
Department of Hepatobiliary Surgery, Second Affiliated Hospital, Chongqing Medical University, Chongqing, 400010, China
关键词:
RNAiNF-κB核糖体蛋白S3肝移植缺血再灌注损伤
Keywords:
RNAi NF-κB ribosomal protein S3 liver transplantation ischemia/reperfusion injury
分类号:
R394-33;R657.3;R364.12
文献标志码:
A
摘要:
目的    采用 RNA干扰技术沉默供体肝脏内核糖体蛋白S3(ribosomal protein S3, RPS3)基因的表达,观察其对核转录因子kappa B(nuclear factor-kappa B,NF-κB)的影响以及对肝移植后早期缺血再灌注损伤的保护作用。    方法    构建以腺病毒(Ad)为载体的RPS3基因特异性短发夹RNA(shRNA)表达颗粒,按随机数字表法将实验大鼠分为3组:Ad-RPS3-shRNA组、空白腺病毒对照组(空白对照组)、生理盐水对照组(NS对照组)。于肝移植前48 h分别用Ad-RPS3-shRNA颗粒、空白腺病毒载体、生理盐水处理3组供体大鼠。收集血液及肝脏组织检测血清转氨酶、血浆炎症因子水平、肝脏组织病理改变、炎症因子mRNA、肝组织提取物中NF-κB活性和表达水平指标的变化。    结果    肝移植后恢复灌注3 h Ad-RPS3-shRNA组大鼠血清ALT水平(725.9±67.3)U/L、AST水平(863.7±72.7)U/L,血浆TNF-α水平(223.7±16.8)ng/L、IL-1β水平(242.5±18.7)ng/L、ICAM-1水平(258.4±24.9)ng/L,Suzuki评分(3.72±0.31),肝组织mRNA水平:TNF-α(0.252±0.026)、IL-1β(0.217±0.028)、ICAM-1(0.226±0.017)、RPS3(0.094±0.007),肝组织NF-κB蛋白水平(0.216±0.022)及其活性(92.4±9.5)均显著低于空白对照组和NS对照组(P<0.01),而空白对照组、NS对照组比较无统计学差异(P>0.05);恢复灌注12 h Ad-RPS3-shRNA组各检测指标与3 h变化趋势相同。    结论    沉默供体肝脏内RPS3基因可显著降低肝脏细胞NF-κB活性和表达量,从而降低炎症因子表达,有效减轻肝移植后早期缺血再灌注损伤。
Abstract:
Objective    To silence ribosome protein S3 (RPS3) gene in donor livers using RNAi technique, and to observe the influence on nuclear factor-kappa B (NF-κB) and the protective effect on early ischemia/reperfusion injury (I/RI) after orthotopic liver transplantation in rats.     Methods    Recombinant adenovirus-RPS3-shRNA granules were constructed. Seventy-two SD rats were randomly divided into three groups: an Ad-RPS3-shRNA group, an Ad-blank-control group, and an NS-control group. Ad-RPS3-shRNA granules, blank adenovirus granules, and normal saline were injected into donor rats in the three groups at 48 h before the donor livers were harvested. The liver tissues and blood samples were harvested in 3 and 12 h after the liver transplantation. Liver function, plasma inflammatory factor level, and liver histopathological change were assessed. Meanwhile, the mRNA levels of inflammatory factors in liver tissues were detected by quantitative RT-PCR. Electrophoretic mobility shift assay and Western blotting were used to assess the activity and level of NF-κB.     Results    At 3 h after reperfusion following liver transplantation, the serum ALT level and serum AST level were 725.9±67.3 U/L and 863.7±72.7 U/L in the Ad-RPS3-shRNA group. The TNF-α, IL-1β and ICAM-1 levels in plasma were 223.7±16.8 ng/L, 242.5±18.7 ng/L and 258.4±24.9 ng/L, while the mRNA levels of TNF-α, IL-1β and ICAM-1 in liver tissues were 0.252±0.026 ng/L, 0.217±0.028 ng/L, and 0.226±0.017 ng/L. Suzuki score was 3.72±0.31, and liver RPS3 mRNA level was 0.094±0.007 ng/L. The liver NF-κB level and activity were 0.216±0.022 and 92.4±9.5, respectively. The above indexes in the Ad-RPS3-shRNA group were significantly lower than those in the Ad-blank-control and the NS-control group (P<0.01). There was no statistical difference between the Ad-blank-control group and the NS-control group (P>0.05). At 12 h after reperfusion following liver transplantation, the above indexes showed the same change tendency as those at 3 h after reperfusion.     Conclusion    Silencing RPS3 gene in donor livers significantly down-regulates the level and activity of NF-κB, inhibits the over-expression of inflammatory factors, and reduces early I/RI after orthotopic liver transplantation.

参考文献/References:

陈新, 成名翔, 龚建平, 等. 沉默供体肝脏内核糖体蛋白S3基因减轻大鼠移植肝脏缺血再灌注损伤[J].第三军医大学学报,2011,33(14):1441-1445.

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更新日期/Last Update: 2011-07-12