羟丁酸钠在大鼠海马脑片缺氧复氧损伤中的保护作用机制

中国临床药理学与治疗学 ›› 2008, Vol. 13 ›› Issue (1): 51-56.

羟丁酸钠在大鼠海马脑片缺氧复氧损伤中的保护作用机制

尤文彬¹, 蒋新颖², 马行¹, 谷淑玲¹, 戴体俊³

¹徐州医学院药理学教研室, ²徐州医学院附属医院检验科, ³江苏省麻醉学重点实验室, 徐州 221002, 江苏

收稿日期:2006-11-10 修回日期:2007-03-21 出版日期:2008-01-26 发布日期:2020-10-13
通讯作者: 谷淑玲, 女, 教授, 硕士生导师, 研究方向:心脑血管药理学。Tel:0516-82771510 E-mail:gushling@163.com
作者简介:尤文彬, 男, 硕士研究生, 研究方向:心脑血管药理学。Tel:0516-85748484 　E-mail:njyou@163.com
基金资助:
国家自然科学基金资助项目(39970715); 江苏省教育厅自然科学基金资助项目(03KJB310141)

Protective effects of sodium oxybate on hypoxia reoxygenation injury of hippocampal slices in rats

YOU Wen-bin¹, JIANG Xin-ying², MA Xing¹, GU Shu-ling¹, DAI Ti-jun³

¹Department of Pharmacology, Xuzhou Medical College, Xuzhou 221002, Jiangsu, China;
²Clinical Laboratory of Affiliated Hospital of Xuzhou Medical College, Xuzhou 221002, Jiangsu, China;
³Jiangsu ProvinceKey Laboratory of Anesthesiology, Xuzhou 221002, Jiangsu, China

Received:2006-11-10 Revised:2007-03-21 Online:2008-01-26 Published:2020-10-13

摘要/Abstract

摘要： 目的研究羟丁酸钠(SO) 在大鼠海马脑片缺氧复氧(H R) 损伤中的保护作用, 揭示SO 在缺血性脑损伤中的保护作用机制。方法采用大鼠海马脑片H R 损伤模型。设正常对照组, H R组, SO 1 、10 、100 μmol/L 组, NCS-382 100 μmol/L +SO 100 μmol/L (NCS-382 + SO) 组、NCS-356100 μmol/L (NCS-356) 组。测定脑片孵育液中乳酸脱氢酶(LDH) 释放率, γ-氨基丁酸(GABA) 、谷氨酸(Glu) 含量;脑片进行TTC 染色计算组织损伤百分率;HE 染色观察组织病理形态学变化, 流式细胞仪测定细胞内钙;酶组织化学法检测一氧化氮合酶(NOS) 的表达。结果 SO 明显降低H R 海马脑片LDH 释放率(P<0.01), 提高TTC 染色的A490值, 降低组织损伤百分率(P<0.01), 升高GABA Glu 比值(P<0.01), 减轻H R 所致的组织病理损伤。H R 组脑片细胞内钙荧光强度和NOS阳性神经元明显高于正常对照组(P<0.01) ;SO 100 μmol/L 组、NCS-356 组细胞内钙荧光强度较H R 组显著减弱(P<0.01),NOS 阳性神经元的数目明显减少(P<0.01)。用γ-羟基丁酸(GHB) 受体选择性阻断剂NCS-382 后再使用SO, 细胞内钙荧光强度、NOS 阳性神经元的数目均接近H R组。结论 SO 对大鼠海马脑片H R 损伤有明显的保护作用, 其机制可能与升高GABA Glu 比值, 激动GHB 受体抑制细胞内钙的增高及NOS 的表达有关。

关键词: 羟丁酸钠, γ-羟基丁酸受体, 缺氧复氧损伤, 细胞内钙, 一氧化氮合酶

Abstract: AIM: To study the protective effects of sodium oxybate(SO) on hypoxia reoxygenation injury of hippocampal slices of rats and investigate its protective mechanism SO to ischemic cerebral injury. METHODS: Experimental model of hypoxia reoxygenation injury of hippocampal slices of rats was adopted. Slices were equally divided into seven groups: control group, hypoxia reoxygenation (H R) group, SO1, 10, 100μmol/L group, NCS-382 [γ-hydroxybutyric acid(GHB) receptors antagonist] 100 μmol/L combined with SO100 μmol/L (NCS-382 +SO100) group, NCS-356 (GHB receptors agonist) 100 μmol/L group. LDH release rate and gamma-aminobutyric acid (GABA) and glutamate (Glu) content in incubation fluid of hippocampal slices were measured. Tissue injury in slices was detected by TTC staining method. The Changes of histomorphology by HE staining were observed as well. Changes of intracellular calcium by flow cytometry and the expression of nitric oxide synthase by enzymohistochemistry method were studied. RESULTS: Hypoxia reoxgenation caused an increase in LDH release rate, a decrease in TTC staining, a decline of GABA Glu ratio and tissue injury obviously, while pretreatment with SO was able to reverse the indexes mentioned above(P<0.01). Fluorescence intensity of intracellular calcium and the number of NOS positive neurons in H R group were higher than those in the control group (P<0.01). However, fluorescence intensity of intracellular calcium (P<0.01) in SO100 and NCS-356 groups was lower than that in H R group, were the number of NOS positive neurons was reduced obviously (P< 0.01). Using SO after GHB receptors antagonist NCS-382, fluorescence intensity of intracellular calcium and the number of NOS positive neurons were similar to HR group. CONCLUSION: SO can protect hippocampal slices subjected to hypoxia reoxygenation injury significantly in rats. The protective mechanism of SO may related to the increase of GABA Glu ratio, the reducing of calcium reflux and decrease of NOS activity by activating GHB receptors.

Key words: sodium oxybate, γ-hydroxybutyric acid receptor, hypoxia reoxygenation injury, intracellular calcium, nitric oxide synthase

中图分类号:

尤文彬, 蒋新颖, 马行, 谷淑玲, 戴体俊. 羟丁酸钠在大鼠海马脑片缺氧复氧损伤中的保护作用机制[J]. 中国临床药理学与治疗学, 2008, 13(1): 51-56.

YOU Wen-bin, JIANG Xin-ying, MA Xing, GU Shu-ling, DAI Ti-jun. Protective effects of sodium oxybate on hypoxia reoxygenation injury of hippocampal slices in rats[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2008, 13(1): 51-56.

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