Inhibition of NAD(P)H oxidase reduces fibronectin expression in stroke-prone renovascular hypertensive rat brain
Inhibition of NAD(P)H oxidase reduces fibronectin expression in stroke-prone renovascular hypertensive rat brain
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DOI：
10.1111/j.1440-1681.2007.04554.x
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发表时间：
2007-04-01
影响因子：
2.9
通讯作者：
Zeng, Jinsheng
中科院分区：
文献类型：
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作者：
Cui, Chunmei;Chen, Alex F.;Zeng, Jinsheng
1. 本研究的目的是验证体内慢性抑制NAD(P)H氧化酶可降低脑卒中易发肾血管性高血压大鼠（RHRSP）脑血管纤维连接蛋白表达的假设。用两种夹子诱导RHRSP模型，并用夹竹桃素抑制NAD(P)H氧化酶。采用实时逆转录聚合酶链反应、western blot和免疫组织化学检测RHRSP大鼠和Sprague-Dawley大鼠脑内NAD(P)H氧化酶亚基p22(phox) mRNA和蛋白的表达。用免疫组织化学方法定位大鼠脑血管组织中纤维连接蛋白的表达，并用免疫印迹法测定其表达量。术后8周，RHRSP组（n = 6）脑血管纤维连接蛋白水平显著高于对照组（n = 5）（分别为1.29 +/- 0.04 vs 1.15 +/- 0.02; P = 0.007）。p22（phox）免疫阳性反应局限于对照大鼠和RHRSP的脑血管系统。此外，用选择性NAD(P)H氧化酶抑制剂罗布宁（apocynin）长期治疗RHRSP 4周（术后5周，1.5 mmol/L），导致p22（phox）蛋白表达显著降低（未治疗的RHRSP为0.85 +/- 0.01 vs 0.93 +/- 0.01, n = 5, P = 0.002），同时脑血管纤维连接蛋白水平降低（未治疗的RHRSP为1.31 +/- 0.03 vs 1.56 +/- 0.05, n = 5, P = 0.002）。RHRSP组（肾动脉收缩后4周和8周）与对照组p22(phox) mRNA和蛋白的表达无显著差异。这些发现提示，罗布麻苷在体内慢性抑制NAD(P)H氧化酶可降低脑血管纤维连接蛋白水平，从而减轻高血压脑血管纤维化。
1. The aim of the present study was to test the hypothesis that in vivo chronic inhibition of NAD(P)H oxidase reduces cerebrovascular fibronectin expression in stroke-prone renovascular hypertensive rats (RHRSP).2. The RHRSP model was induced by two clips and NAD(P)H oxidase was inhibited with apocynin. The mRNA and protein expression of NAD(P)H oxidase subunit p22(phox) in brains of RHRSP and Sprague-Dawley (control) rats was determined using real-time reverse transcription-polymerase chain reaction, western blot and immunohistochemistry. The expression of fibronectin protein was localized immunohistochemically in cerebral vessels and then quantified by western blot.3. Cerebrovascular fibronectin levels in RHRSP (n = 6) were significantly higher than control (n = 5) levels 8 weeks after operation (1.29 +/- 0.04 vs 1.15 +/- 0.02, respectively; P = 0.007). The p22(phox) immunopositive reactivity was localized in the cerebral vasculature of control rats and RHRSP. Furthermore, chronic treatment of RHRSP with apocynin, a selective NAD(P)H oxidase inhibitor, in the drinking water for 4 weeks (1.5 mmol/L, 5 weeks after operation) resulted in a significant decrease in the expression of p22(phox) protein (0.85 +/- 0.01 vs 0.93 +/- 0.01 in non-treated RHRSP; n = 5; P = 0.002), with a concomitant reduction of fibronectin levels in the cerebral vasculature (1.31 +/- 0.03 vs 1.56 +/- 0.05 in non-treated RHRSP; n = 5; P = 0.002). No significant differences were detected in the expression of p22(phox) mRNA and protein between RHRSP (4 and 8 weeks after renal artery constriction) and the control group.4. These findings suggest that the chronic inhibition of NAD(P)H oxidase in vivo by apocynin reduces cerebrovascular fibronectin levels, which may lessen hypertensive cerebrovascular fibrosis.