整合素连接激酶在低氧诱导肺动脉高压发病机制中的作用

鄢高亮; 盛祖龙; 周千星; 袁春菊; 潘啸东; 陈中璞; 汤成春; 马根山

doi:10.13201/j.issn.1001-1439.2017.02.019

整合素连接激酶在低氧诱导肺动脉高压发病机制中的作用

- 东南大学附属中大医院心内科(南京, 210009)
基金项目:
国家自然科学基金(No:81600227,81370225)

详细信息

通讯作者: 鄢高亮,E-mail:yanshipingguo@163.com

中图分类号: R544.1

收稿日期: 2016-09-19

修回日期: 2016-12-13

Effect of integrin linked kinase in the pathogenesis of hypoxia-induced pulmonary hypertension in rats

- Department of Cardiology, Zhongda Hospital of Southeast University Medical School, Nanjing, 210009, China

More Information

Corresponding author: YAN Gaoliang, E-mail: yanshipingguo@163.com

Received Date: 19 September 2016

Revised Date: 13 December 2016

摘要

摘要: 目的:通过了解整合素连接激酶(ILK)及心肌素(myocardin)在低氧性肺动脉高压(PAH)大鼠肺组织及肺血管中的表达,探讨ILK在低氧性肺动脉高压发病机制中的作用。方法:48只SD大鼠按随机数字表法分为常氧对照组和低氧1周、2周、4周组,低氧干预大鼠置于低氧仓内,氧浓度控制在(10.5±0.5)％,每日间断低氧8 h(8:00~16:00)。测定各组大鼠肺动脉平均压(mPAP)、计算右室肥厚指数[右室/(左室+室间隔),RV/(LV+S)]、管壁厚度占血管外径的百分比(WT％)和管壁面积占血管总面积的百分比(WA％);采用髓鞘碱性蛋白为底物通过液体闪烁仪测定ILK活性;实时定量PCR法测定肺动脉内ILK、糖原合成激酶3β(GSK-3β)及myocardin mRNA相对表达量;Western印迹法检测肺动脉及培养的肺动脉平滑肌细胞(PASMC)胞质内ILK、GSK-3β及myocardin蛋白相对表达量。结果:低氧2周组mPAP、右室肥厚指数、WT%、WA%均显著高于常氧对照组(均P＜0.05),低氧4周组差异更为显著。低氧1周组肺动脉内ILK活性即明显下降达36%(P＜0.05),低氧2周、4周组ILK活性均较常氧对照组明显降低(均P＜0.05)。低氧4周组ILK mRNA和蛋白相对表达量均显著低于常氧对照组(均P＜0.05),myocardin mRNA和蛋白表达变化与ILK相似(均P＜0.05),而GSK-3β的mRNA和蛋白表达量则呈相反趋势,显著高于常氧对照组(均P＜0.05)。低氧培养PASMC 24 h后ILK及myocardin蛋白表达明显低于常氧对照组,而GSK-3β蛋白水平则呈相反趋势,低氧可促进其表达增加(均P＜0.05)。结论:慢性低氧引起ILK活性和表达下调,使其下游靶点GSK-3β表达上调,进而降低myocardin表达水平,可能是导致PAH形成的重要机制。
- 整合素连接激酶 /
- 高血压 /
- 肺性 /
- 缺氧 /
- 大鼠
Abstract: Objective:To explore the expression of integrin linked kinase and myocardin in the pathogenesis of hypoxia-induced pulmonary artery hypertension (PAH) in a rat model.Method:Forty-eight Sprague-Dawley rats were randomly divided into 4 groups (n=12 each). And the rats were exposed to normoxia or hypoxia for 1,2,4 weeks respectively. SD rats were exposed to chronic intermittent hypoxia [PO₂:(10.5±0.5)%, 8:00-16:00]. Mean pulmonary arterial pressure (mPAP),right ventricular hypertrophy (RV/(LV+S)) and the index of wall thickness of small pulmonary artery (WT% and WA%) among groups were measured. The kinase activity of ILK was measured through myelin basic protein substrate by liquid scintillation counting. Real-time quantitative Polymerase Chain Reaction was performed to quantify the relatively mRNA levels of ILK, glycogen synthase kinase-3 beta(GSK-3β), and myocardin. And western blot assay were employed to determine the relative expressions of proteins of ILK, GSK-3β and myocardin. Result:The levels of mPAP,RV/(LV+S),WT% and WA% were significantly higher after 2 week hypoxia than those in normoxic control(all P＜0.05). And the levels of above in 4 weeks were further increased compared with normoxic control. The kinase activity of ILK was significantly decreased at 36%(P＜0.05) in 1 week after hypoxia compared with normoxic control, and the levels of kinase activity in 2, 4 weeks were also reduce compared with normoxic control(39%,43%,all P＜0.05).The relative expression of ILK mRNA and protein were significantly decreased after 4 week hypoxia than that in normoxic control(all P＜0.05).And the similar variation tendency were observed in myocardin(all P＜0.05).However, the relative expression of GSK-3β mRNA and protein were significantly increased after 4 week hypoxia than that in normoxic control(all P＜0.05).In vitro, the relative protein expression of ILK and myocardin were significantly reduced by hypoxia for 24 h compared with normoxia treatment in cultured pulmonary artery smooth muscle cells. However, the protein level of GSK-3β were dramatically increased by hypoxia treatment.Conclusion:The down regulation of the kinase activity and expression of ILK under chronic hypoxia may further induce the up regulation of GSK-3β, followed by downregulation of myocardin, which is probably implicated in the pathogenesis of PAH.
- integrin linked kinase /
- hypertension /
- pulmonary /
- hypoxia /
- rats

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参考文献(10)

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