【摘要】 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。更多还原

【Abstract】 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.更多还原