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目的:探讨不同浓度的谷红注射液对乳鼠海马神经元因缺氧缺糖(OGD)所造成损伤的保护作用。方法:培养原代乳鼠海马神经元并用神经元特异性烯醇化酶(NSE)免疫组化染色法鉴定。将培养成熟的细胞随机分成6组:正常组、OGD模型组、尼莫地平组(200g/m L)、谷红注射液低、中、高剂量(30、60、90g/m L)组,除正常组之外,余组都加药并进行OGD处理,造模后检测各组乳鼠海马神经元细胞中丙二醛(MDA)、一氧化氮(NO)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-PX)表达活性及细胞上清液乳酸脱氢酶(LDH)外漏量的变化情况,采用酶联免疫吸附法检测各组细胞上清液中白细胞介素-6(IL-6)以及单核细胞趋化蛋白-1(MCP-1)含量,采用流式细胞术检测谷红注射液对各组海马神经元细胞凋亡的影响。结果:鉴定表明,海马神经元纯度>90%,可用于实验。OGD使原代培养的海马神经元受损,与OGD模型组相比,谷红注射液高、中、低剂量组可以有效减轻缺氧造成的损伤,能够有效提高SOD与GSH-PX活性,减轻MDA表达,减少LDH外漏以及NO含量,减弱MCP-1与IL-6表达,抑制海马神经元凋亡(P<0.05,P<0.01)。结论:谷红注射液可以减轻新生乳鼠海马神经元OGD损伤,其保护机制可能与增强细胞抗氧化应激能力、减轻炎性反应与抗细胞凋亡有关。
Objective: To investigate the protective effects of different concentrations of Gu-hong injection on damage of hippocampal neurons induced by oxygen and glucose deprivation (OGD). Methods: Primary cultured neonatal rat hippocampal neurons were cultured and identified by neuron specific enolase (NSE) immunohistochemical staining. The cultured mature cells were randomly divided into 6 groups: normal group, OGD model group, nimodipine group (200g / m L), Guhong injection low, medium and high doses (30,60,90 g / m L) Except for the normal group, the rest of the groups were dosed with OGD and treated with OGD. MDA, NO, SOD (supersonic superoxide dismutase) ) And glutathione peroxidase (GSH-PX) expression and cell supernatant lactate dehydrogenase (LDH) leakage were measured by enzyme-linked immunosorbent assay in each group of cell supernatant (IL-6) and monocyte chemoattractant protein-1 (MCP-1) in hippocampal neurons were detected by flow cytometry. Results: Identification showed that hippocampal neurons purity> 90%, can be used for experiments. Compared with OGD model group, OGD can damage primary cultured hippocampal neurons. Compared with OGD model group, high, medium and low doses of Guhong injection can effectively reduce the damage caused by hypoxia, and can effectively improve the activity of SOD and GSH-PX, reduce (P <0.05, P <0.01). The expression of MCP-1 and IL-6 in the hippocampal neurons were decreased after the treatment with LDH. Conclusion: Gu-Hong injection can reduce OGD injury in hippocampal neurons of neonatal neonatal rats. Its protective mechanism may be related to its ability to enhance anti-oxidative stress and reduce inflammatory reaction and anti-apoptosis.