PARP抑制剂对蛛网膜下腔出血后迟发型血管痉挛及炎症因子的影响

The effect of PARP inhibitor on the delayed development cerebral vasospasm after subarachnoid hemorrhage and inflammatory factors

目的研究多聚腺苷二磷酸核糖聚合酶(PARP)抑制剂3-氨基苯甲酰胺(3-AB)对大鼠蛛网膜下腔出血(SAH)后迟发型脑血管痉挛(DCVS)的作用及对炎症因子单核细胞趋化蛋白-1(MCP-1)、超敏C反应蛋白(hsCRP)含量的影响,并探讨其与NF-κB信号通路的关系。方法将80只SD大鼠随机分为正常对照组(n=8)、假手术组(n=8)、SAH组(n=32)和3-AB(n=32)组。采用枕大池2次注血法建立大鼠SAH模型,并给与大鼠腹腔注射3-AB(30mg/kg),于2次注血后3、5、7、14d处死取材,光镜下观察SAH后脑基底动脉形态变化;ELISA检测脑组织中PARP和MCP-1、hsCRP的含量;免疫组化方法检测大鼠脑基底动脉NF-κB的表达。结果与假手术组比较,SAH模型建立后第5天,大鼠基底动脉痉挛程度达到峰值[(30.47±3.89)%],基底动脉管壁厚度和管腔内径分别为(16.44±1.32)μm和(178.21±11.13)μm,脑血流量减少近60%(P<0.01),基底动脉NF-κB在胞质及核内表达明显增强,脑组织中PARP含量显著升高(P<0.01),MCP-1、hsCRP含量亦显著增高(P<0.01),分别为(365.29±28.08)pg/mL和(402.16±48.99)ng/mL;与SAH组比较,3-AB干预5d后,大鼠基底动脉痉挛程度、管腔狭窄、管壁增厚明显缓解(P<0.01),其值分别为(22.65±3.21)%、(14.89±1.27)μm和(198.56±10.91)μm,脑血流量明显增加,基底动脉NF-κB在胞质及核内表达明显降低,脑组织中PARP的含量显著降低(P<0.01),脑组织MCP-1、hsCRP含量显著下降(P<0.01),分别为(126.51±18.67)pg/mL和(285.39±39.07)ng/mL。结论PARP抑制剂3-AB能够减轻SAH后迟发性脑血管痉挛,抑制脑组织的炎症反应,其机制可能与NF-κB信号通路有关。

Objective To investigate the effects of poly ADP-ribose polymerase （PARP） inhibitor ：3- aminobenzene （3-AB） on the delayed development cerebral vasospasm （DCVS） after subarachnoid hemorrhage （SAH） and on the inflammatory factors, namely monocyte chemotactic protein 1 （MCP-1） and hypersensitive c-reactive protein （hsCRP）, and to explore the relationship between these and the signaling pathway of NF-kappa B （NF-kB）. Methods Eighty male Sprague-Dawley rats were randomly divided into four groups： normal group （n = 8）, sham-operation group （n = 8）, SAH model group （n = 32） and 3-AB group （n = 32）. We established 64 SAH model animals by double injection of blood into the cisterna magna. Half of the SAH model animals were treated with 3-AB by intraperitoneal injection （30 mg/kg）. These rats were killed to obtain specimens respectively at days 3, 5, 7 and 14 after the second blood injection. The morphological changes of basilar arteries were observed under thc light microscope. The contents of PARP, MCP-1 and hsCRP in brain tissues were detected with enzyme-linked immunosorbent assay （ELISA）. The expression of NF-kB in basilar arteries was determined by immunohistochcmistry. Results Compared with those in the sham-operation group, the degree of basilar artery spasm reached the peak [-（30.47±3.89）%] at day 5 after established SAH model; the thickness and diameter of basilar artery were （16.44±1.32）μm and （178. 21± 11.13）μm, respectively. Cerebral blood flow was reduced by nearly 60% （P〈0.01）. The expression of NF-KB in the cytoplasm and nucleus and PARP content in brain tissue were both increased significantly （P〈0. 01）. MCP-I [（365. 29±28. 08）pg/mL] and hsCRP [（402. 16 ±48. 99）ng/mL] were significantly enhanced （P〈0.01）. Compared with the SAH group, after 5 days＇ intervention with 3-AB, there was obvious alleviation in the spasm degree of basilar artery [ （22.65±3.21 ）% , the thickness [ （14.89±1.27）μm] and diameter [ （ 198.56± 10.91） μm], respectively （P〈 0.01 ）. Cerebral blood flow was significantly enhanced, but the expression of NF-kB in the cytoplasm and nucleus was decreased and PARP in brain tissue was significantly decreased （P〈0.01）. MCP-1 [（126.51±18. 67）pg/mL] and hsCRP [（285. 39 ± 39. 07）ng/mL] in brain tissue were significantly declined, respectively （P〈0.01）. Oonclusion PARP inhibitor 3-AB can alleviate DCVS and inhibit the inflammatory response in brain tissue after SAH. The mechanism may be related to NF-kB signaling pathway.