百里醌对大鼠蛛网膜下腔出血后脑细胞凋亡的影响
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摘要
目的研究百里醌对大鼠蛛网膜下腔出血(subarachnoid hemorrhage,SAH)后脑细胞凋亡的作用及其机制。方法采用血管内穿刺法建立SAH模型,将48只成年雄性SD大鼠采用数字表法随机分为假手术组(Sham组)、SAH组、媒介组和百里醌组,SAH造模24h后检测神经功能损伤、脑组织含水率和脑组织内依文蓝(EB)含量;Tunel法检测脑细胞凋亡;ELISA法检测脑组织中IL-13的表达;Western blot法检测脑组织中IL-13Rα2、p-PI_3K、p-AKT和m TOR的表达。结果 SAH组大鼠Garcia神经功能评分、脑组织含水率和EB含量较Sham组均明显升高,脑组织中IL-13的表达上调;SAH组大鼠脑出血后细胞凋亡率显著上升,IL-13Rα2、p-PI_3K、p-AKT和m TOR的表达升高;与媒介组相比较,百里醌组大鼠Garcia神经功能评分、脑组织含水率和EB含量均明显降低,IL-13在脑组织中的表达下调,细胞凋亡率显著下降,IL-13Rα2、p-PI_3K、p-AKT和m TOR的表达下调。结论百里醌可减轻大鼠SAH后脑损伤和脑细胞凋亡,其机制可能通过IL-13Rα2/PI_3K/AKT/m TOR通路实现。
Objective To explore the effects of thymoquinone on early brain injury and apoptosis in rats after subarachnoid hemorrhage( SAH). Methods Total 48 adult male SD rats were randomly divided in to Sham group,SAH group,vehicle group and thymoquinone group. Neurological score,brain water content and blood-brain barrier were evaluated at 24 h after SAH. Tunel staining was performed to detect cell apoptosis in rat brain tissue. ELISA method was performed for quantitative detection of IL-13. The relative levels of IL-13 Rα2,p-PI3 K,p-AKT and m TOR in rat brain tissue were detected by western blot. Results 24 h after modeling,Garcia neurological score,brain water content and EB content of brain tissue of rats in SAH group were significantly higher than that of Sham group.ELISA analysis revealed that SAH induced a significant increase in IL-13 expression in the brain tissue. Tunel staining showed that the number of TUNEL-stained cells was increased in the subcortical brain region after SAH compared to the sham group. In addition,a higher expression of IL-13 Rα2,p-PI3 K,p-AKT and m TOR in rat brain were observed at 24 h post-SAH. However,the neurological deficit scores,brain water content and EB content in rat brains were significantly reduced in thymoquinone group in comparison with vehicle group. The administration of thymoquinone dramatically suppressed the level of IL-13 in the rat brains. Thymoquinone treatment grossly reduced the number of TUNEL positive cells. Furthermore,thymoquinone significantly decreased the levels of IL-13 Rα2,p-PI3 K,p-AKT as well as m TOR. Conclusion Thymoquinone could effectively inhibit early brain injury and apoptosis following SAH in rats,which may be related to down-regulation of IL-13 Rα2/PI_3 K/AKT/m TOR pathway.
Objective To explore the effects of thymoquinone on early brain injury and apoptosis in rats after subarachnoid hemorrhage( SAH). Methods Total 48 adult male SD rats were randomly divided in to Sham group,SAH group,vehicle group and thymoquinone group. Neurological score,brain water content and blood-brain barrier were evaluated at 24 h after SAH. Tunel staining was performed to detect cell apoptosis in rat brain tissue. ELISA method was performed for quantitative detection of IL-13. The relative levels of IL-13 Rα2,p-PI3 K,p-AKT and m TOR in rat brain tissue were detected by western blot. Results 24 h after modeling,Garcia neurological score,brain water content and EB content of brain tissue of rats in SAH group were significantly higher than that of Sham group.ELISA analysis revealed that SAH induced a significant increase in IL-13 expression in the brain tissue. Tunel staining showed that the number of TUNEL-stained cells was increased in the subcortical brain region after SAH compared to the sham group. In addition,a higher expression of IL-13 Rα2,p-PI3 K,p-AKT and m TOR in rat brain were observed at 24 h post-SAH. However,the neurological deficit scores,brain water content and EB content in rat brains were significantly reduced in thymoquinone group in comparison with vehicle group. The administration of thymoquinone dramatically suppressed the level of IL-13 in the rat brains. Thymoquinone treatment grossly reduced the number of TUNEL positive cells. Furthermore,thymoquinone significantly decreased the levels of IL-13 Rα2,p-PI3 K,p-AKT as well as m TOR. Conclusion Thymoquinone could effectively inhibit early brain injury and apoptosis following SAH in rats,which may be related to down-regulation of IL-13 Rα2/PI_3 K/AKT/m TOR pathway.
引文
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16 庄宗,赵旭东,吴颐,等.PI3K-AKT信号通路在蛛网膜下腔出血后的抗凋亡作用[J].医学研究生学报,2010,6(23):579-582
17 Tu M,Wange W,Cai L,et al.IL-13 receptorα2 stimulates human glioma cell growth and metastasis through the Src/PI3K/AKT/m TOR signaling pathway[J].Tumour Biol,2016,37(11):14701-14709
2 Turek G,Lewszuk A,Kochanowicz J,et al.Early outcomes and perioperative complications of endovascular embolization in patients with aneurysmal SAH[J].Neurol Neurochir Pol,2016,50(5):342-348
3 Chu F,Borthakur A,Sun X,et al.The Siva-1 putative amphipathic helical region(SAH)is sufficient to bind to BCL-XL and sensitize cells to UV radiation induced apoptosis[J].Apoptosis,2004,9(1):83-95
4 Liu H,Yang M,Pan L,et al.Hyperbaric oxygen intervention modulates early brain injury after experimental subarachnoidhemorrhage in rats:possible involvement of TLR4/NF-x03BA;B-mediated signaling pathway[J].Cell Physiol Biochem,2016,38(6):2323-2336
5 Karaca G,Aydin O,Pehlivanli F,et al.Effectiveness of thymoquinone,zeolite,and platelet-rich plasma in model of corrosive oesophagitis induced in rats[J].Ann Surg Treat Res,2017,92(6):396-401
6 Alobaedi OH,Talib WH,Basheti IA.Antitumor effect of thymoquinone combined with resveratrol on mice transplanted with breast cancer[J].Asian Pac J Trop Med,2017,10(4):400-408
7 Mahmud HA,Seo H,Kim S,et al.Thymoquinone(TQ)inhibits the replication of intracellular mycobacterium tuberculosis in macrophages and modulates nitric oxide production[J].BMC Complement Altern Med,2017,17(1):279
8 Bederson JB,Germano IM,Guarino L.Cortical blood flow and cerebral perfusion pressure in a new noncraniotomy model of subarachnoid hemorrhage in the rat[J].Stroke,1995,26(6):1086-1091
9 Gu H,Fei ZH,Wang YQ,et al.Angiopoietin-1 and Angiopoietin-2 Expression Imbalance Influence in Early Period After Subarachnoid Hemorrhage[J].Int Neurourol J,2016,20(4):288-295
10 Tao C,Fan C,Hu X,et al.The effect of fenestration of the lamina terminalis on the incidence of shunt-dependent hydrocephalus after aneurysmal subarachnoid hemorrhage(FISH):study protocol for a randomized controlled trial[J].Medicine(Baltimore),2016,95(52):e5727
11 Jin Q,Cai Y,Li S,et al.Edaravone-encapsulated agonistic micelles rescue ischemic brain tissue by tuning blood-brain barrier permeability[J].Theranostics,2017,7(4):884-898
12 Fuss IJ,Joshi B,Yang Z,et al.IL-13Rα2-bearing,typeⅡNKT cells reactive to sulfatide self-antigen populate the mucosa of ulcerative colitis[J].Gut,2014,63(11):1728-1736
13 Joshi BH,Puri RK.IL-13 receptor-alpha2:a novel target for cancer therapy[J].Immunotherapy,2009,1(3):321-327
14 Sivaprasad U,Warrier MR,Gibson AM,et al.IL-13Rα2 has a protective role in a mouse model of cutaneous inflammation[J].J Immunol,2010,185(11):6802-6808
15 刘俊杰,李建民,赵雅宁,等.PI3K-m TOR信号通路对蛛网膜下腔出血大鼠海马区神经细胞自噬的调控作用[J].西安交通大学学报:医学版,2017,38(2):188-192
16 庄宗,赵旭东,吴颐,等.PI3K-AKT信号通路在蛛网膜下腔出血后的抗凋亡作用[J].医学研究生学报,2010,6(23):579-582
17 Tu M,Wange W,Cai L,et al.IL-13 receptorα2 stimulates human glioma cell growth and metastasis through the Src/PI3K/AKT/m TOR signaling pathway[J].Tumour Biol,2016,37(11):14701-14709