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  • 孔二亮,陈默,拜云虎,邵金鹏,凤旭东.瑞芬太尼通过激活PI3K/Akt/eNOS通路减轻肾缺血/再灌注损伤[J].第二军医大学学报,2019,40(12):1337-1343    [点击复制]
  • KONG Er-liang,CHEN Mo,BAI Yun-hu,SHAO Jin-peng,FENG Xu-dong.Remifentanil alleviates renal ischemia/reperfusion injury by activating PI3K/Akt/eNOS pathway[J].Acad J Sec Mil Med Univ,2019,40(12):1337-1343   [点击复制]
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瑞芬太尼通过激活PI3K/Akt/eNOS通路减轻肾缺血/再灌注损伤
孔二亮1,陈默2,拜云虎3,邵金鹏4,凤旭东1*
0
(1. 中国人民解放军联勤保障部队 988 医院麻醉科, 郑州 450042;
2. 海军军医大学(第二军医大学)东方肝胆外科医院麻醉科, 上海 200438;
3. 中国人民解放军联勤保障部队 988 医院肝胆外科, 郑州 450042;
4. 中国人民解放军联勤保障部队988医院泌尿外科, 郑州 450042
*通信作者)
摘要:
目的 探讨术中应用瑞芬太尼对肾缺血/再灌注损伤的作用及机制。方法 取雄性C57/BL小鼠50只,随机分为5组:假手术组、缺血/再灌注损伤组(I/R组)、缺血/再灌注损伤+磷脂酰肌醇3-激酶(PI3K)抑制剂LY294002处理组(I/R+LY294002组)、缺血/再灌注损伤+瑞芬太尼处理组(I/R+RF组)、缺血/再灌注损伤+LY294002处理+瑞芬太尼处理组(I/R+RF+LY294002组),每组10只。各组大鼠分别于手术结束6 h后取静脉血或肾脏组织,采用全自动生化分析仪测定静脉血尿素氮(BUN)、血肌酐(SCr)水平,采用蛋白质印迹法检测肾脏组织PI3K/蛋白激酶B(Akt)/内皮型一氧化氮合酶(eNOS)通路相关蛋白的表达,H-E染色观察肾脏组织炎症细胞聚集情况,酶联免疫吸附试验测定肾脏组织中肿瘤坏死因子α(TNF-α)、白细胞介素(IL)-1β、IL-6、IL-10等炎性因子的表达,实时定量PCR测定肾脏组织中抗凋亡因子Bcl2和凋亡因子半胱氨酸蛋白酶3(Caspase-3)的mRNA表达。结果 与假手术组相比,I/R组小鼠静脉血BUN、SCr水平均升高,肾脏组织中PI3K表达及磷酸化Akt(p-Akt)/Akt、磷酸化eNOS(p-eNOS)/eNOS均降低,炎性因子TNF-α、IL-1β、IL-6、IL-10释放均增加,Bcl2 mRNA表达下降而Caspase-3 mRNA表达增高,差异均有统计学意义(P均<0.05),并且肾脏组织中炎症细胞聚集增多。给予瑞芬太尼处理后,I/R+RF组BUN、SCr水平较I/R组降低,肾脏组织中PI3K表达及p-Akt/Akt、p-eNOS/eNOS均增高,TNF-α、IL-1β、IL-6、IL-10释放均减少,Bcl2 mRNA表达上调、Caspase-3 mRNA表达下调,与I/R组相比差异均有统计学意义(P均<0.05),炎症细胞募集也减轻。而使用PI3K抑制剂LY294002处理后,I/R+RF+LY294002组BUN、SCr水平升高,肾脏组织中p-eNOS/eNOS降低,IL-1β、L-6释放增加,Caspase-3 mRNA表达上调,与I/R+RF组相比差异均有统计学意义(P均<0.05),炎症细胞募集也增加。结论 肾缺血/再灌注损伤时,瑞芬太尼通过激活PI3K/Akt/eNOS通路减轻肾脏组织炎症反应和细胞凋亡发挥肾脏保护作用。
关键词:  瑞芬太尼  磷脂酰肌醇3-激酶  蛋白激酶B  内皮型一氧化氮合酶  肾缺血/再灌注损伤  炎症反应  细胞凋亡
DOI:10.16781/j.0258-879x.2019.12.1337
投稿时间:2019-07-01修订日期:2019-09-18
基金项目:
Remifentanil alleviates renal ischemia/reperfusion injury by activating PI3K/Akt/eNOS pathway
KONG Er-liang1,CHEN Mo2,BAI Yun-hu3,SHAO Jin-peng4,FENG Xu-dong1*
(1. Department of Anesthesiology, No. 988 Hospital of Logistic Support Forces of PLA, Zhengzhou 450042, Henan, China;
2. Department of Anesthesiology, Eastern Hepatobiliary Surgery Hospital, Naval Medical University (Second Military Medical University), Shanghai 200438, China;
3. Department of Hepatobiliary Surgery, No. 988 Hospital of Logistic Support Forces of PLA, Zhengzhou 450042, Henan, China;
4. Department of Urology, No. 988 Hospital of Logistic Support Forces of PLA, Zhengzhou 450042, Henan, China
*Corresponding author)
Abstract:
Objective To explore the effect and mechanism of intraoperative remifentanil (RF) on renal ischemia/reperfusion (I/R) injury. Methods Fifty male C57/BL mice were randomly divided into 5 groups:sham group, I/R group, I/R+LY294002 (a phosphatidylinositol 3-kinase[PI3K] inhibitor) group, I/R+RF group and I/R+RF+LY294002 group, with 10 mice in each group. Venous blood or renal tissue samples were collected from the mice of each group 6 h after I/R operation. The levels of blood urea nitrogen (BUN) and serum creatinine (SCr) were detected using automatic biochemical analyzer. The expression levels of PI3K/protein kinase B (Akt)/endothelial nitric oxide synthase (eNOS) pathway-related proteins in renal tissues of mice were detected using Western blotting. The aggregation of inflammatory cells was observed by H-E staining. The expression levels of tumor necrosis factor α (TNF-α), interleukin (IL)-1β, IL-6 and IL-10 in renal tissues of mice were detected by enzyme-linked immunosorbent assay. The mRNA expression levels of anti-apoptotic factor Bcl2 and apoptotic factor Caspase-3 in renal tissues were determined by real-time quantitative PCR. Results Compared with the sham group, the BUN and SCr levels in venous blood were increased in the I/R group, the PI3K expression, phosphorylated-Akt (p-Akt)/Akt ratio and phosphorylated-eNOS (p-eNOS)/eNOS ratio in renal tissues were decreased, the release levels of inflammatory factors (TNF-α, IL-1β, IL-6 and IL-10) were increased, Bcl2 mRNA expression was decreased, and Caspase-3 mRNA expression was increased; and the differences were significant (all P<0.05). The mice of the I/R group had increased inflammatory cell recruitment in renal tissues. After RF treatment, the mice of the I/R+RF group had decreased levels of BUN and SCr in venous blood, increased PI3K expression, p-Akt/Akt ratio and p-eNOS/eNOS ratio in renal tissues, decreased release levels of inflammatory factors (TNF-α, IL-1β, IL-6 and IL-10), increased Bcl2 mRNA expression, and decreased Caspase-3 mRNA expression; and the differences were significant compared with the mice of the I/R group (all P<0.05). The inflammatory cell recruitment was decreased in the I/R+RF group. Moreover, compared with the mice of the I/R+RF group, the mice of the I/R+RF+LY294002 group had increased levels of BUN and SCr in venous blood, decreased p-eNOS/eNOS ratio in renal tissues, increased IL-1β and IL-6 release, and increased Caspase-3 mRNA expression; and the differences were significant (all P<0.05). The inflammatory cell recruitment was increased in the I/R+RF+LY294002 group. Conclusion RF exerts protective effect on kidney with I/R injury by alleviating renal inflammation and cell apoptosis through activating PI3K/Akt/eNOS pathway.
Key words:  remifentanil  phosphatidylinositol 3-kinase  protein kinase B  endothelial nitric oxide synthase  renal ischemia/reperfusion injury  inflammation  apoptosis