[缺氧诱导因子-1α对大鼠心肌缺血/再灌注损伤的保护作用及蛋白激酶C在信号通路中的作用]

[Protective effects of hypoxia-inducible factor-1 alpha on myocardial ischemia/reperfusion injury in rat and the role of protein kinase C in signal pathway].

作者信息

Niu Tie-sheng, Qi Guo-xian, Fu Peng, Sun Ying-xian

机构信息

Department of Cardiology, the Shengjing Hospital of China Medical University, Shenyang 110004, Liaoning, China.

出版信息

Zhongguo Wei Zhong Bing Ji Jiu Yi Xue. 2010 Feb;22(2):101-4.

PMID:20170615

Abstract

OBJECTIVE

To study the expression of hypoxia-inducible factor-1 alpha (HIF-1 alpha) in a rat model of myocardial ischemia/reperfusion injury (IRI) and the role of protein kinase C (PKC) in signal pathway.

METHODS

A rat model of myocardial IRI was reproduced by 30 minutes of left anterior descending coronary artery (LCA) occlusion followed by 180 minutes of reperfusion. Thirty-two healthy male Wistar rats were randomly divided into four groups. The first group was ischemic preconditioning (IPC) group; the second group was simple IRI group; the third group was IPC plus PKC inhibitor group (IPC+I group); the fourth group was the sham-operation group without ligation of LCA. Eight rats were used in each group. The heart was harvested 180 minutes post-reperfusion, the mRNA and protein expression of HIF-1 alpha and heme oxygenase-1 (HO-1) were assessed. Meanwhile, the protein expression of caspase-3 was assayed. Blood samples were obtained from heart to determine the levels of interleukin-8 (IL-8) and myeloperoxidase (MPO).

RESULTS

The mRNA and protein expression of HIF-1 alpha and HO-1 increased significantly in the IRI group compared with the sham-operation group, while the protein expression of caspase-3 increased significantly in the IRI group (HIF-1 alpha mRNA: 0.849+/-0.032 vs. 0.356+/-0.022, HIF-1 alpha protein: 0.762+/-0.042 vs. 0.324+/-0.016, HO-1 mRNA: 0.862+/-0.045 vs. 0.332+/-0.012, HO-1 protein: 0.792+/-0.044 vs. 0.335+/-0.031, caspase-3 protein: 0.371+/-0.015 vs. 0.061+/-0.012, respectively, all P<0.01). The levels of IL-8 and MPO increased significantly in the IRI group [IL-8: (812+/-26) ng/L vs. (72+/-13) ng/L, MPO: (78.7+/-2.9) kU/L vs. (13.3+/-1.5) kU/L, both P<0.01]. The protein and mRNA expression of HIF-1 alpha and HO-1 increased significantly in the IPC group compared with IRI group (HIF-1 alpha mRNA: 1.412+/-0.039, HIF-1 alpha protein: 1.362+/-0.045, HO-1 mRNA: 1.523+/-0.038, HO-1 protein: 1.420+/-0.041, respectively), meanwhile the protein expression of caspase-3 (0.129+/-0.019) decreased significantly in the IPC group (all P<0.01). The levels of IL-8 [(432+/-59) ng/L] and MPO [(43.2+/-5.9) kU/L] decreased significantly in the IPC group compared with IRI group (both P<0.01). All above parameters showed no significant change between IPC+I group and IRI group.

CONCLUSION

HIF-1 alpha plays a protective role in myocardial IRI, PKC is an important signal pathway of HIF-1 alpha gene expression in IRI.

摘要

目的

研究缺氧诱导因子-1α（HIF-1α）在大鼠心肌缺血/再灌注损伤（IRI）模型中的表达以及蛋白激酶C（PKC）在信号通路中的作用。

方法

采用冠状动脉左前降支（LCA）阻断30分钟后再灌注180分钟的方法建立大鼠心肌IRI模型。32只健康雄性Wistar大鼠随机分为四组。第一组为缺血预处理（IPC）组；第二组为单纯IRI组；第三组为IPC加PKC抑制剂组（IPC+I组）；第四组为未结扎LCA的假手术组。每组8只大鼠。再灌注180分钟后取心脏，检测HIF-1α和血红素加氧酶-1（HO-1）的mRNA和蛋白表达。同时，检测半胱天冬酶-3的蛋白表达。从心脏取血样测定白细胞介素-8（IL-8）和髓过氧化物酶（MPO）水平。

结果

与假手术组相比，IRI组HIF-1α和HO-1的mRNA和蛋白表达显著增加，而IRI组半胱天冬酶-3的蛋白表达显著增加（HIF-1α mRNA：0.849±0.032对0.356±0.022，HIF-1α蛋白：0.762±0.042对0.324±0.016，HO-1 mRNA：0.862±0.045对0.332±0.012，HO-1蛋白：0.792±0.044对0.335±0.031，半胱天冬酶-3蛋白：0.371±0.015对0.061±0.012，均P<0.01）。IRI组IL-8和MPO水平显著升高[IL-8：（812±26）ng/L对（72±13）ng/L，MPO：（78.7±2.9）kU/L对（13.3±1.5）kU/L，均P<0.01]。与IRI组相比，IPC组HIF-1α和HO-1的蛋白和mRNA表达显著增加（HIF-1α mRNA：1.412±0.039，HIF-1α蛋白：1.362±0.045，HO-1 mRNA：1.523±0.038，HO-1蛋白：1.420±0.041），同时IPC组半胱天冬酶-3的蛋白表达（0.129±0.019）显著降低（均P<0.01）。与IRI组相比，IPC组IL-8[（432±59）ng/L]和MPO[（43.2±5.9）kU/L]水平显著降低（均P<0.01）。上述所有参数在IPC+I组和IRI组之间均无显著变化。