栀子苷对大鼠蛛网膜下腔出血后早期脑损伤氧化应激的保护作用
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1.广州中医药大学第二附属医院 脑病中心,广州 510120; 2.广州中医药大学第二附属医院 儿科, 广州 510120; 3.广州中医药大学第二临床医学院,广州 510120; 4.南京大学医学院附属鼓楼医院 神经外科, 南京 210008

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R-33


Geniposide protects against oxidative stress in early bRain injury after subarachnoid hemorrhage
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Affiliation:

1.Diagnosis and Treatment Center of Encephalopathy, the 2nd Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine, Guangzhou 510120, China. 2. Department of Pediatrics, the 2nd Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine, Guangzhou 510120. 3. the Second Clinical School of Guangzhou University of Chinese Medicine, Guangzhou 510120. 4. Department of Neurosurgery, Nanjing Drum Tower Hospital, School of Medicine, Nanjing University, Nanjing 210008

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    摘要:

    目的 本研究探讨栀子苷(GNP)对大鼠蛛网膜下腔出血( SAH)后早期脑损伤(EBI)氧化应激的保护作用。 方法 采用视交叉前池注血法制备蛛网膜下腔出血大鼠模型,将雄性 Wistar 大鼠随机分为假手术组、模型组、GNP 低、高剂量组,造模后一次腹腔内注射。 24 h 后进行蛛网膜下腔出血量评分及死亡率计算;免疫荧光染色检测颞叶皮层脑组织核因子 2 相关因子 2( nuclear factor erythroiD-2-related factor 2,Nrf2)表达情况;Western blot 测定颞叶皮层血红素加氧酶-1(hemeoxygenase-1,HO-1)、醌氧化还原酶-1(NADPH quinineoxidoreductase-1,NQO-1)、谷胱甘肽-S-转移酶(glutathione S-transferase,GST)蛋白含量;TBA 比色法检测颞叶皮层中丙二醛(malondialdehyde, MDA)含量,TUNEL 染色法检测神经细胞凋亡情况。 结果 与假手术组比较,模型组大鼠死亡率百分比增加;颞叶皮层 Nrf2 阳性细胞数目、HO-1、NQO-1、GST 蛋白水平略有增加;MDA 水平显著升高、大量细胞发生凋亡(P<0. 05)。 与模型组比较,GNP 两剂量组大鼠颞叶皮层 Nrf2 阳性细胞数目、HO-1、NQO-1、GST 蛋白水平均显著增加( P< 0. 05),MDA 水平,细胞凋亡率明显下降(P<0. 05);其中高剂量组死亡率百分比并出现下降。 结论 本研究发现 GNP 对大鼠 SAH 后 EBI 具有一定保护作用,其作用机制可能与通过激活内源性抗氧化系统 Nrf2 通路,上调下游抗氧化酶发挥对神经细胞的保护作用密切相关。

    Abstract:

    Objective This study aimed to test the hypothesis that geniposide(GNP) protects against oxidative stress in early bRain injury(EBI)after experimental subarachnoid hemorrhage(SAH). Methods Male Wistar rats(n= 48) were randomly allocated to four groups, including the sham + vehicle group, SAH + vehicle group, SAH +GNP low dose group, and SAH + GNP high dose group. The experimental SAH model was established by injecting blood into the prechiasmatic cistern and treatments were then administered. Outcomes were measured at 24 hours after treatment. The SAH score and mortality rate were calculated. Nuclear factor erythroiD-2-related factor 2(Nrf2)expression in the temporal cortex was detected by immunofluorescence staining. Heme oxygenase-1(HO-1), NADPH quinineoxidoreductase-1(NQO-1), and glutathione S-transferase ( GST ) protein expression in the temporal cortex was determined by western blotting. Malondialdehyde(MDA)levels in the temporal cortex were detected by the thiobarbituric acid method . Apoptosis of nerve cells was determined using TUNEL staining. Results In the SAH+vehicle group, MDA levels and the rate of cellular apoptosis were significantly higher compared with those in the sham+vehicle group( both P<0. 05). The number of Nrf2- positive cells and HO-1, NQO-1, and GST expression were significantly higher in the SAH+vehicle group compared with the sham+vehicle group(all P<0. 05). Compared with the SAH+vehicle group, Ttreatment of either a high or low dose of GNP result ed in a higher number of Nrf2-positive cells and higher HO-1, NQO-1, and GST expression, and lower MDA levels and cellular apoptosis rate(all P<0. 05). In addition, Llower mortality was also observed in the SAH+GNP high dose group. group treated with a high dose of GNP(100 mg / kg). Conclusions GNP exerts a protective effect against oxidative stress in EBI after SAH. This effect is probably mediated by activating the Nrf2 pathway and upregulating the downstream of pathway—antioxidants.

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黎劭学,肖晓岚,尹 雷,孟苗苗,岑学程,蔡 军,杭春华.栀子苷对大鼠蛛网膜下腔出血后早期脑损伤氧化应激的保护作用[J].中国比较医学杂志,2021,31(4):55~62.

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  • 收稿日期:2020-07-30
  • 在线发布日期: 2021-05-28
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