Salt-inducible kinase 2 regulates energy metabolism in rats with cerebral ischemia-reperfusion

doi:10.3724/zdxbyxb-2021-0164

J Zhejiang Univ (Med Sci)

2021, Vol. 50

Issue (3): 352-360 DOI: 10.3724/zdxbyxb-2021-0164

Salt-inducible kinase 2 regulates energy metabolism in rats with cerebral ischemia-reperfusion

ZHANG Ran¹²(

),LIU Yun¹²,ZHANG Cui¹,MA Mengyao¹,LI Shu^12,*(

),HONG Yun^3,*(

)

1. Department of Pathophysiology, Wannan Medical College, Wuhu 241002, Anhui Province, China;
2. Clinical Colloge of Wannan Medical College, Wuhu 241002, Anhui Province, China; 3. Department of Ultrasonic Medicine, Yijishan Hospital of Wannan Medical College, Wuhu 241001, Anhui Province, China

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Abstract

Objective:To investigate the effects of salt-inducible kinase 2 (SIK2) on energy metabolism in rats with cerebral ischemia-reperfusion. Methods: Adult SD male rats (240-260?g) were divided into 5 groups: sham group, ischemia group, reperfusion group, adenovirus no-load group, and SIK2 overexpression group with 5 animals in each group. The middle cerebral artery occlusion (MCAO) was induced with the modified Zea-Longa line thrombus method to establish the cerebral ischemia reperfusion model. Eight days before the MCAO, SIK2 overexpression was induced by injecting 7 μL adenovirus in the right ventricle, then MCAO was performed for 2?h, followed by reperfusion 24?h. HE staining was used to observe the pathological changes of cerebral tissue in rats; TTC staining was used to observe the volume of cerebral infarct. The levels of adenosine triphosphate (ATP) and adenosine diphosphate (ADP) in rat brain tissue were detected by ELISA; the levels of SIK2 and hypoxia-inducible factor 1α (HIF-1α) in the rat brain tissues were detected by RT-qPCR and Western blotting. Results:Compared with the sham group, SIK2 level was decreased in the ischemia group, and it was further declined in the reperfusion group (P<0.05). Compared with the sham group and ischemic group, the pathological injury in reperfusion group were more severe, and the infarct size was larger; compared with the reperfusion group and adenovirus no-load group, the pathological injury of the SIK2 overexpression group was milder, and the infarct size is less. Compared with the sharn group, HIF-1α was increased in both ischemia group and reperfusion group, especially in ischemia group (allP<0.05); HIF-1α level in the SIK2 overexpression group was higher than that in the reperfusion group and adenovirus no-load group (allP<0.05). ATP level in ischemia group and reperfusion group was lower than that in the sham group, and the reperfusion group decreased more significantly than the ischemia group (P<0.05); ADP content was increased in the ischemia and reperfusion group, and the ADP content in reperfusion group was significantly higher than that in the ischemia group (P<0.05). ATP level in the SIK2 overexpression group was higher than that in the reperfusion group and adenovirus no-load group (allP<0.05), and ADP was decreased in the SIK2 overexpression group (allP<0.05).Conclusion:SIK2 can up-regulate the ATP level and down-regulate the ADP level in rat brain tissue and alleviate cerebral ischemia-reperfusion injury by increase the level of HIF-1α.

Key words： Ischemia reperfusion Energy metabolism Salt-inducible kinase 2 Hypoxia-inducible factor-1α Adenosine triphosphate Adenosine diphosphate SD rat

Received: 11 January 2021 Published: 16 August 2021

CLC:

R363

Corresponding Authors: LI Shu,HONG Yun E-mail: 18255366693@163.com;yxx2003@126.com

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	ZHANG Ran
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Cite this article:

ZHANG Ran,LIU Yun,ZHANG Cui,MA Mengyao,LI Shu,HONG Yun. Salt-inducible kinase 2 regulates energy metabolism in rats with cerebral ischemia-reperfusion. J Zhejiang Univ (Med Sci), 2021, 50(3): 352-360.

URL:

http://www.zjujournals.com/med/10.3724/zdxbyxb-2021-0164 OR http://www.zjujournals.com/med/Y2021/V50/I3/352

盐诱导激酶2对脑缺血再灌注大鼠脑组织能量代谢的影响

目的：探究盐诱导激酶2（SIK2）对脑缺血再灌注大鼠能量代谢相关物质表达的影响。方法：选用成年SD雄性大鼠（240~260?g），参照改良Zea-Longa线栓法建立大鼠短暂性中动脉栓塞模型（MCAO），在构建MCAO模型前8?d予以右侧脑室注射7?μL腺病毒使大鼠脑组织SIK2过表达，随后建立缺血2?h再灌注24?h模型。实验分为手术对照组、缺血对照组、缺血再灌注组、腺病毒空载组及SIK2过表达组。苏木精-伊红（HE）染色观察大鼠神经细胞损伤的病理学改变；氯化三苯基四氮唑（TTC）染色观察大鼠脑组织梗死情况；酶联免疫吸附试验（ELISA）检测各组大鼠脑组织中腺苷三磷酸（ATP）、腺苷二磷酸（ADP）的含量；荧光定量聚合酶链反应和蛋白质印迹法检测各组大鼠脑组织中SIK2及缺氧诱导因子1α（HIF-1α）的表达量。结果：与手术对照组比较，缺血对照组及缺血再灌注组SIK2表达减少，且缺血再灌注组较缺血对照组减少更明显（P<0.05）；与手术对照组和缺血对照组比较，缺血再灌注组病理损伤较重，梗死体积较大；与缺血再灌注组和腺病毒空载组比较，SIK2过表达组病理损伤较轻，梗死体积减少。与手术对照组比较，缺血对照组和缺血再灌注组HIF-1α表达均增加，其中缺血对照组增加更明显（均P<0.05）；SIK2过表达组HIF-1α表达比缺血再灌注组和腺病毒空载组均增多（均P<0.05）。与手术对照组比较，缺血对照组和缺血再灌注组ATP含量均减少，且缺血再灌注组较缺血对照组减少更为显著（P<0.05）；ADP含量在缺血对照组和缺血再灌注组均增多，且缺血再灌注组较缺血对照组明显增多（P<0.05）；与缺血再灌注组和腺病毒空载组比较，SIK2过表达组ATP含量增加（均P<0.05），ADP含量减少（均P<0.05）。结论：SIK2可通过上调HIF-1α的表达，增加大鼠脑组织中ATP的含量，减少ADP含量，减轻大鼠脑缺血再灌注损伤。

关键词： 缺血再灌注, 能量代谢, 盐诱导激酶2, 缺氧诱导因子1α, 腺苷三磷酸, 腺苷二磷酸, SD大鼠

Figure 1 Electrophoretogram of SIK2 expression in brain tissues of sham group, ischemia group and ischemia-reperfusion group

Table 1 Comparison of SIK2 levels among brain tissues of sham group, ischemia group and ischemia-reperfusion group

Figure 2 HE staining of brain tissues in each groups

Figure 3 Infarction of brain tissues in each group

Figure 4 Electro phoretogram of HIF-1α expression in each group

Table 2 The level of HIF-1α in ischemia and reperfusion rats

Table 3 The level of HIF-1α in ischemic-reperfusion rat brain tissue with SIK2 overexpression

Table 4 The content of ATP and ADP in brain tissues of each group


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