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
作者:
作者单位:

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

作者简介:

李智勇,海南医学院第一附属医院医疗质量管理科,邮箱:lizy0060@163.com。

通信作者:

彭俊,邮箱:pengj060@163.com。

基金项目:

海南省自然科学基金青年项目(821QN422)。


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

1.Medical Quality Management Department, The First Affiliated Hospital of Hainan Medical College, Haikou, Hainan 570102, China;2.Department of Neurosurgery, The First Affiliated Hospital of Hainan Medical College, Haikou Hainan 570102, China;3.Department of Neurosurgery, Haikou Affiliated Hospital of Central South University Xiangya School of Medicine, Haikou, Hainan 570208, China

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    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。

    Abstract:

    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 model of SAH was established by intravascular perforation. Intraventricular injection of Lv-GPR30 was performed in the rats with SHA. 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 Western blotting and real-time fluorescence quantitative PCR, respectively. ELISA was employed to measure the levels of tumor necrosis 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 SAH and improved their neurological function, neuroinflammation, BBB disruption, and cerebral edema. Overexpression of GPR30 inhibited the expression of matrix metallopeptidase-9 and matrix metallopeptidase-2 as well as the inflammatory factors TNF-α, IL-6, and IL-1β in the brain tissue of SAH rats but increased the IL-10 level.Conclusions GPR30 can alleviate neuroinflammation and BBB disruption in rats with SAH.

    表 4 各组大鼠BWC和BBB通透性的比较Table 4
    表 5 各组大鼠脑组织中MMP-9及MMP-2的表达量比较Table 5
    表 2 各组大鼠脑组织中GPR30的表达量比较Table 2
    Fig.
    Fig.
    图3 各组大鼠脑组织中GPR30的表达水平Fig.3
    Fig.
    表 1 各组大鼠脑组织中GPR30蛋白的表达量比较Table 1
    表 6 各组大鼠脑组织中IL-6、IL-1β、TNF-α、IL-10的水平比较Table 6
    表 3 各组大鼠的Garcia评分比较Table 3
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李智勇,陈政纲,彭俊456. 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[J].国际神经病学神经外科学杂志,2024,(2):29-34111LI Zhiyong, CHEN Zhenggang, PENG Jun222. GPR30 attenuates neuroinflammation and blood-brain barrier disruption in rats with subarachnoid hemorrhage[J]. Journal of International Neurology and Neurosurgery,2024,(2):29-34

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  • 收稿日期:2023-11-05
  • 最后修改日期:2024-04-12
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  • 在线发布日期: 2024-06-19
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