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目的观察尼古丁对脂多糖(LPS)诱导的小胶质细胞激活及活化后细胞死亡的影响。方法建立慢性尼古丁暴露的小鼠动物模型,腹腔注射LPS诱导小胶质细胞激活,应用免疫组织化学方法观察皮质、海马、黑质CD-11b阳性小胶质细胞表达的变化;BV2细胞(小鼠小胶质瘤细胞系)传代培养,运用CCK-8试剂盒检测细胞活性,一氧化氮检测试剂盒检测一氧化氮(NO)释放情况,RT-PCR分析诱导型一氧化氮合酶(iNOS)、肿瘤坏死因子(TNF-α)、白细胞介素1(IL-1β)、白细胞介素6(IL-6)、环氧化酶-2(COX-2)、干扰素调节因子1(IRF-1)、Caspase-11mRNA的表达,免疫印迹法分析P-I-κB、Caspase-3的表达变化。结果尼古丁抑制LPS诱导的皮质、海马、黑质CD-11b阳性小胶质细胞的表达;尼古丁抑制LPS刺激引起的BV2细胞的死亡,NO的释放,iNOS、TNF-αI、L-1βI、L-6、COX-2I、RF-1、Caspase-11 mRNA的表达,P-I-κB、Caspase-3蛋白的表达。结论尼古丁可以抑制LPS诱导的小胶质细胞活化及激活诱导的细胞死亡(AICD),对脑内炎症反应具有神经保护作用。
Objective To observe the effects of nicotine on lipopolysaccharide (LPS) -induced microglial activation and cell death after activation. Methods The animal model of chronic nicotine exposure was established. The microglia were activated by intraperitoneal injection of LPS. The expression of CD-11b positive microglial cells in cortex, hippocampus and substantia nigra was observed by immunohistochemistry. Microglioma cell lines) were subcultured. Cell viability was measured by CCK-8 kit. Nitric oxide (NO) release was measured by nitric oxide kit. Inducible nitric oxide synthase (iNOS) , TNF-α, IL-1β, IL-6, COX-2 and IRF- 1), Caspase-11mRNA expression, Western blot analysis of PI-κB, Caspase-3 expression changes. Results Nicotine inhibited the expression of CD-11b positive microglial cells induced by LPS in cortex, hippocampus and substantia nigra. Nicotine inhibited the death of BV2 cells induced by LPS, the release of NO, the expression of iNOS, TNF-αI, L-1βI, L- 6, COX-2I, RF-1, Caspase-11 mRNA expression, PI-κB, Caspase-3 protein expression. Conclusion Nicotine can inhibit LPS-induced microglial activation and activation-induced cell death (AICD), which has neuroprotective effects on inflammatory reaction in the brain.