白藜芦醇对实验性兔肺动脉高压相关炎症因子表达的影响

Effect of resveratrol on expression of inflammatory factors related to pulmonary arterial hypertension in rabbits

摘要:

目的  通过检测白藜芦醇（Res）干预后实验性兔肺动脉高压（PAH）模型肺组织中相关细胞因子表达水平的差异，探讨其在PAH发病机制中的作用。方法  将36只新西兰兔随机分为6组。对照组（A组）：动物皮下注射二甲基亚砜30 mg/kg。造模组：实验组动物适应性饲养1周后，皮下注射野百合碱30 mg/kg，连续注射7 d造模，然后将造模动物随机分为5组：模型组（B组）、实验药物组（C组）、高剂量组（D1组）、中剂量组（D2组）、低剂量组（D3组），每组6只。注射后第45天，分别进行B超、心肌细胞苏木精-伊红染色和Tunnel染色，检测造模是否成功，造模成功后按体重计算给药剂量，受试药物高剂量120 mg/（kg·d），中剂量60 mg/（kg·d），低剂量30 mg/（kg·d），阳性对照药物前列腺环素1 mg/（kg·d）喂饲相应的动物，1次/d，连续6周，每日观察动物的食量、活动、毛色及大小便变化，每周称体重。每隔2周随机采集各组实验动物外周血样本，采用酶联免疫吸附法试剂盒，分别对实验动物血清3种生物学标志物[转录因子-κB（NF-κB）、环氧化酶-2（COX-2）、诱导型一氧化氮合酶（iNOS）]的含量进行检测。结果  Res干预实验性兔PAH模型后，NF-κB、COX-2、iNos血清表达水平改善，其中高剂量组改善效果优于阳性对照药物前列腺环素。且在实验中经对比发现，NF-κB的特异性最明显，其他指标虽然有变化，但显著性不如NF-κB。结论  Res抑制血清中分泌NF-κB、COX-2、iNos的能力优于前列腺环素，Res可以通过抑制NF-κB/iNOS/COX2通路避免细胞损伤，改善PAH症状，表现出和前列腺环素相同或更好的作用。

Abstract:

Objective To investigate the effect of resveratrol on the expression of related cytokines in lung tissue of pulmonary arterial hypertension (PAH) rabbits, and to explore their roles in the pathogenesis of PAH. Methods Thirty-six New Zealand rabbits were randomly divided into six groups. In the model control group (group A), the rabbits were subcutaneously injected with DMSO (30 mg/kg). To build the pulmonary arterial hypertension (PAH) model, after one week of adaptive feeding in the experimental group, the rabbits had subcutaneous injection of monocrotaline (MCT, 30 mg/kg) for 7 consecutive days. Then the PAH model animals were randomly divided into 5 groups: model group (group B), positive drug control group (group C), high-dose group (group D1), midium-dose group (group D2), low-dose group (group D3) with 6 in each group. On the 45th day after injection, ultrasonic examination and myocardial cell observation after HE staining and tunnel staining were used to make sure the success of modeling. After successful modeling, the dosage of drug was calculated according to body weight. The rabbits in the groups D1, D2 and D3 were fed with 120 mg/(kg·d) (high dose), 60 mg/(kg·d) (midium dose) and 30 mg/(kg·d) (low dose) of resveratrol respectively; prostacyclin 1 mg/(kg·d) was used to feed the rabbits in the positive drug control group, once a day for 6 week. The changes of food intake, activity, hair color, and urine and feces were observed daily, the rabbits were weighed weekly. Peripheral blood samples were randomly collected from each group every 2 weeks. ELISA kits were used to detect serum levels of transcription factor kappa B (NF-κB), cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) respectively. Results The serum levels of NF-κB, COX-2 and iNOS were significantly improved after resveratrol intervention in the PAH rabbits, and the effect in the high-dose group was better than that in the positive control group. And among the three indicators NF-κB had the most obvious specificity. Conclusions Resveratrol inhibits serum levels of NF-κB, COX-2 and, iNOS, which is better than that of the prostacyclin. Resveratrol can prevent cell damage and improve symptoms of PAH by inhibiting the NF-κB/iNOS/COX-2 pathway, which is the same as or better than the effect of prostacyclin.