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中华诊断学电子杂志 ›› 2018, Vol. 06 ›› Issue (03) : 202 -206. doi: 10.3877/cma.j.issn.2095-655X.2018.03.013

所属专题： 文献；

基础研究

7，8-二羟基黄酮对自发性高血压大鼠血管平滑肌细胞表型转化的影响
姚天成1, 王伟杰1, 杨莹莹1, 郑祥珍1, 刘海青1, 王炳香1,†()   
  1. 1. 271000　泰安，泰山医学院基础医学院生理学教研室
  • 收稿日期:2018-02-08 出版日期:2018-08-26
  • 通信作者: 王炳香
  • 基金资助:
    国家自然科学基金(81500363); 国家大学生创新创业训练计划项目(201610439028); 山东省自然科学基金(ZR2017LC009); 山东省高等学校科技计划(J17KA138); 泰安市科技局引导项目(2016NS1060)

Effect of 7, 8-dihydroxyflavone on the phenotype transformation of vascular smooth muscle cells in spontaneously hypertensive rats

Tiancheng Yao1, Weijie Wang1, Yingying Yang1, Xiangzhen Zheng1, Haiqing Liu1, Bingxiang Wang1,†()   

  1. 1. Department of Physiology, Basic Medical College of Taishan Medical University, Tai′an 271000, China
  • Received:2018-02-08 Published:2018-08-26
  • Corresponding author: Bingxiang Wang
  • About author:
    Corresponding author: Wang Bingxiang, Email:
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引用本文：

姚天成, 王伟杰, 杨莹莹, 郑祥珍, 刘海青, 王炳香. 7，8-二羟基黄酮对自发性高血压大鼠血管平滑肌细胞表型转化的影响[J/OL]. 中华诊断学电子杂志, 2018, 06(03): 202-206.

Tiancheng Yao, Weijie Wang, Yingying Yang, Xiangzhen Zheng, Haiqing Liu, Bingxiang Wang. Effect of 7, 8-dihydroxyflavone on the phenotype transformation of vascular smooth muscle cells in spontaneously hypertensive rats[J/OL]. Chinese Journal of Diagnostics(Electronic Edition), 2018, 06(03): 202-206.

目的

探讨7，8-二羟基黄酮(7，8-DHF)对自发性高血压大鼠(SHR)血管平滑肌细胞(VSMCs)表型转化的影响。

方法

用Western blotting法检测清洁级雄性Wistar-Kyoto(WKY)大鼠及SHR胸主动脉和VSMCs中平滑肌肌动蛋白(SM-α-actin)及增殖细胞核抗原(PCNA)的蛋白表达水平；用不同浓度7，8-DHF和(或)TrkB特异性抑制剂ANA-12处理SHR胸主动脉源性VSMCs，用CCK-8法检测VSMCs活性；用Western blotting法检测7，8-DHF和(或)ANA-12处理SHR胸主动脉源性VSMCs后SM-α-actin和PCNA的蛋白表达水平。使用Prism 5统计软件，应用独立样本t检验或单因素方差分析进行统计学分析。

结果

同WKY大鼠比较，SHR胸主动脉组织中SM-α-actin的蛋白表达水平显著降低[(0.72±0.06)，(0.41±0.08)；t＝6.35，P<0.05]，PCNA的蛋白表达水平则明显升高[(0.51±0.09)，(0.99±0.15)；t＝6.43，P<0.05]。SHR胸主动脉源性VSMCs中SM-α-actin的蛋白表达水平较WKY大鼠显著降低[(0.71±0.05)，(0.36±0.04)；t＝11.12，P<0.01], PCNA的蛋白表达水平则明显升高[(0.69±0.07)，(1.05±0.08)；t＝8.49，P<0.05]。7，8-DHF能明显抑制SHR胸主动脉源性VSMCs增殖活性，并呈浓度依赖性(F＝17.49，P<0.01)，给予ANA-12(10μmol/L)处理后，SHR胸主动脉源性VSMCs的增殖活性较单独给予7，8-DHF组显著升高[(0.52±0.08)，(0.86±0.14)；t＝5.13，P<0.01]；ANA-12能够显著消除7，8-DHF引起的SHR胸主动脉源性VSMCs中SM-α-actin蛋白表达水平的升高[(0.85±0.13)，(0.35±0.15)；t＝4.37，P<0.01]及PCNA蛋白表达水平的降低[(0.42±0.13)，(0.76±0.10)；t＝3.54，P<0.05]。

结论

7，8-DHF能够有效抑制SHR胸主动脉源性VSMCs由收缩型向增殖型转化，其作用可能是由TrkB受体介导。

关键词: 7，8-二羟基黄酮, 高血压, 血管平滑肌细胞, 表型, 受体，trkB
Objective

To investigate the effect of 7, 8-dihydroxyflavone (7, 8-DHF) on the phenotypic transformation of vascular smooth muscle cells (VSMCs) in spontaneously hypertensive rats (SHR).

Methods

Western blotting assay was carried out to detect the Wistar-Kyoto rats and SHR′s protein expression of smooth muscle actin (SM-α-actin) and proliferating cell nuclear antigen (PCNA). With the different concentrations of 7, 8-DHF and (or) TrkB specific inhibitor ANA-12, the VSMCs derived from thoracic aortic of SHR were processed and the VSMCs activity was detected by CCK-8 method. Western blotting was used to detect the protein expression levels of SM-α-actin and PCNA after 7, 8-DHF and (or) ANA-12 treated in VSMCs derived from thoracic aortic of SHR. The independent sample t-test and single factor variance analysis were used for statistical comparison.

Results

Compared with WKY, the expression of SM-α-actin in aorta of SHR decreased significantly [(0.72±0.06), (0.41±0.08), t=6.35, P<0.05], while PCNA increased[(0.51±0.09), (0.99±0.15), t=6.43, P<0.05], and so was the case in VSMCs derived from thoracic aortic of SHR, SM-α-actin decreased [(0.71±0.05), (0.36±0.04), t=11.12, P<0.01] and PCNA increased[(0.69±0.07), (1.05±0.08), t=8.49, P<0.05]. The proliferative activity of VSMCs derived from thoracic aortic of SHR was greatly attenuated by 7, 8-DHF in a concentration-dependent manner(F=17.49, P<0.01), which could be enhanced by ANA-12 [ (0.52±0.08), (0.86±0.14), t=5.13, P<0.01]. ANA-12 can significantly weaken the effect of 7, 8-DHF on the expression of SM-α-actin[(0.85±0.13), (0.35±0.15), t=4.37, P<0.01]and PCNA protein[(0.42±0.13), (0.76±0.10), t=3.54, P<0.05]in VSMCs derived from thoracic aortic of SHR.

Conclusion

7, 8-DHF can effectively inhibit the transformation of VSMCs derived from thoracic aortic of SHR from contractile to proliferative type, which may be mediated by TrkB receptor.

Key words: 7, 8-dihydroxyflavone, Hypertension, Vascular smooth muscle cell, Phenotype, Receptor, trkB
图1 WKY大鼠及SHR胸主动脉和VSMCs中SM-α-actin和PCNA蛋白免疫印迹图
图2 WKY大鼠及SHR胸主动脉源性VSMCs镜下图像(× 20)
图3 各组SHR胸主动脉源性VSMCs中SM-α-actin和PCNA蛋白免疫印迹图
表1 各组SHR胸主动脉源性VSMCs中SM-α-actin和PCNA蛋白表达水平的比较(±s)
组别 SM-α-actin PCNA
normal组 0.81±0.09 0.63±0.14
con组 0.36±0.11 0.97±0.19
DHF组 0.85±0.13 0.42±0.13
DHF＋ANA-12组 0.35±0.15 0.76±0.10
F值 15.40 7.98
P值 <0.01 <0.01
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