G蛋白偶联受体30在大鼠蛛网膜下腔出血后对神经炎症和血脑屏障破坏的影响  

GPR30 attenuates neuroinflammation and blood-brain barrier disruption in rats with subarachnoid hemorrhage

作　　者：李智勇 陈政纲 彭俊[3] LI Zhiyong;CHEN Zhenggang;PENG Jun(Medical Quality Management Department,The First Affiliated Hospital of Hainan Medical College,Haikou,Hainan 570102,China;Department of Neurosurgery,The First Affiliated Hospital of Hainan Medical College,Haikou Hainan 570102,China;Department of Neurosurgery,Haikou Affiliated Hospital of Central South University Xiangya School of Medicine,Haikou,Hainan 570208,China)

机构地区：[1]海南医学院第一附属医院医疗质量管理科,海南海口570102 [2]海南医学院第一附属医院神经外科,海南海口570102 [3]中南大学湘雅医学院附属海口医院神经外科,海南海口570208

出　　处：《国际神经病学神经外科学杂志》2024年第2期29-34,共6页Journal of International Neurology and Neurosurgery

基　　金：海南省自然科学基金青年项目(821QN422)。

摘　　要：目的探讨G蛋白偶联受体30(GPR30)在大鼠蛛网膜下腔出血(SAH)早期脑损伤(EBI)过程中对神经炎症和血脑屏障(BBB)破坏的影响。方法36只雄性大鼠随机分为6组(n=6/组):假手术(Sham)组,SAH(3 h、6 h、12 h、24 h、72 h)组。此外,72只大鼠随机分为4组(n=18/组):Sham组、SAH组、SAH联合过表达GPR30慢病毒阴性载体(SAH+Lv-NC)组、SAH联合过表达GPR30慢病毒载体(SAH+Lv-GPR30)组。通过血管内穿孔建立SAH模型,于SHA大鼠脑室内注射Lv-GPR30。通过神经学评分、脑组织含水量(BWC)检测、伊文思蓝(EBP)染色、苏木精和伊红(HE)染色来分析GPR30对EBI的影响;采用蛋白质印迹法(Western blotting)和实时荧光定量PCR(qRT-PCR)分析各种蛋白质和转录水平;通过酶联免疫吸附测定法(ELISA)分别测定肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、白细胞介素1β(IL-1β)、白细胞介素10(IL-10)水平。结果SAH大鼠脑内注射Lv-GPR30后脑组织中GPR30的表达增加,并改善了大鼠神经功能、神经炎症、BBB破坏和脑水肿程度。过表达GPR30抑制SAH大鼠脑组织中基质金属蛋白肽酶9(MMP-9)和基质金属蛋白肽酶2(MMP-2)的表达,以及炎症因子TNF-α、IL-6、IL-1β表达水平,同时提高IL-10的表达水平。结论GPR30能减轻SAH大鼠的神经炎症和BBB破坏。Objective To investigate the effect of G protein-coupled receptor 30(GPR30)on neuroinflammation and blood-brain barrier(BBB)disruption during early brain injury(EBI)in rats with subarachnoid hemorrhage(SAH).Methods Thirty-six male rats were randomly divided into six groups(n=6 per group):sham operation(Sham)group and SAH(3 h,6 h,12 h,24 h,72 h)groups.In addition,72 rats were randomly divided into four groups(n=18 per group):Sham group,SAH group,SAH combined with overexpressed GPR30 lentivirus negative vector(SAH+Lv-NC) group, SAH combined with overexpressed GPR30 lentivirus vector (SAH+Lv-GPR30) group. A Sprague-Dawley rat modelof SAH was established by intravascular perforation. Intraventricular injection of Lv-GPR30 was performed in the rats withSHA. The effect of GPR30 on EBI was analyzed through neurological scoring, brain tissue water content measurement,Evans blue staining, and HE staining. The levels of proteins and their transcription levels were determined by Westernblotting and real-time fluorescence quantitative PCR, respectively. ELISA was employed to measure the levels of tumornecrosis factor-alpha (TNF- α), interleukin-6 (IL-6), interleukin-1 beta (IL-1β), and interleukin-10 (IL-10).Results Intraventricular injection of Lv-GPR30 increased the expression of GPR30 in the brain tissue of rats with SAHand improved their neurological function, neuroinflammation, BBB disruption, and cerebral edema. Overexpression ofGPR30 inhibited the expression of matrix metallopeptidase-9 and matrix metallopeptidase-2 as well as the inflammatoryfactors TNF-α, IL-6, and IL-1β in the brain tissue of SAH rats but increased the IL-10 level. Conclusions GPR30 canalleviate neuroinflammation and BBB disruption in rats with SAH.

关 键 词：蛛网膜下出血 G蛋白偶联受体30 早期脑损伤 血脑屏障 神经炎症 

分 类 号：R743[医药卫生—神经病学与精神病学]