白杨素对野百合碱诱导的肺动脉高压大鼠右心室NOX4及NF-κB表达的影响
Effect of chrysin on expression of NOX4 and NF-κB in right ventricle of monocrotaline-induced pulmonary arterial hypertension of rats
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摘要: 观察白杨素(chrysin, 5,7-二羟基黄酮)对野百合碱(monocrotaline, MCT)诱导的肺动脉高压(pulmonaryarterial hypertension, PAH)大鼠右心室重构的影响及其机制。Sprague-Dawley大鼠适应性喂养1周,随机分为正常对照组、MCT组、MCT+chrysin (50 mg·kg-1·d-1)及MCT+chrysin (100 mg·kg-1·d-1)剂量组。MCT (60 mg·kg-1)皮下注射诱导PAH大鼠模型。连续给药4周后,右颈外静脉插管测定大鼠右心室收缩压(RVSP)、平均肺动脉压(mPAP)。分离大鼠右心室(RV)、左心室+室间隔(LV+S)并称重,剥离大鼠胫骨并测量其长度,计算RV/(LV+S)及RV/胫骨长度的比值。HE染色观察右心室病理学变化, Masson染色观察右心室胶原沉积的变化。比色法测定右心室总抗氧化能力(T-AOC)和丙二醛(MDA)含量。qPCR、Western blot和(或)免疫组化检测右心室collagen I、collagen III、NADPH氧化酶4(NADPH oxidase 4, NOX4)、核转录因子κB (nuclear factor-kappa B, NF-κB) mRNA和蛋白表达。结果发现,白杨素连续给药4周后能明显降低MCT诱导的PAH大鼠RVSP及mPAP,减轻RV/(LV+S)及RV/胫骨长度的比值,改善右心室病理变化,降低右心室胶原的沉积及collagen I、collagen III的表达,提高右心室T-AOC水平,降低NOX4的表达及MDA含量,抑制NF-κB的表达。结果表明,白杨素能缓解MCT诱导的PAH大鼠右心室重构,其机制可能与其抑制NOX4介导的氧化应激损伤,进而抑制NF-κB所介导的胶原沉积有关。Abstract: The aim of the present study is to investigate the protective effect of chrysin (5,7-dihydroxyflavone) on right ventricular remodeling in a rat model of monocrotaline-induced pulmonary arterial hypertension (PAH). PAH rats were induced by a single injection of monocrotaline (60 mg·kg-1, sc) and were administered with chrysin (50 or 100 mg·kg-1·d-1) for 4 weeks. At the end of experiment, the right ventricular systolic pressure (RVSP) and mean pulmonary artery pressure (mPAP) were monitored via the right jugular vein catheterization into the right ventricle. Right ventricle (RV) to left ventricle (LV) + septum (S) and RV to tibial length were calculated. Right ventricular morphological change was observed by HE staining. Masson 's trichrome stain was used to demonstrate collagen deposition. The total antioxidative capacity (T-AOC) and malondialdehyde (MDA) levels in right ventricle were determined according to the manufacturer 's instructions. The expressions of collagen I, collagen III, NADPH oxidase 4 (NOX4) and nuclear factor-kappa B (NF-κB) were analyzed by immunohistochemisty, qPCR and (or) Western blot. The results showed that chrysin treatment for 4 weeks attenuated RVSP, mPAP and right ventricular remodeling index (RV/LV+S and RV/Tibial length) of PAH rats induced by monocrotaline. Furthermore, monocrotaline-induced right ventricular collagen accumulation and collagen I and collagen III expression were both significantly suppressed by chrysin. The expressions of NOX4, NF-κB and MDA contents were obviously decreased, while the T-AOC was significantly increased in right ventricule from PAH rats with chrysin treatment. These results suggest that chrysin ameliorates right ventricular remodeling of PAH induced by monocrotaline in rats through its down-regulating of NOX4 expression and antioxidant activity, and inhibiting NF-κB expression and collagen accumulation.
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