CAJ | 학술논문

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-11 mRNA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-α、IL-1β、IL-6、COX-2、IRF-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.
목적 관찰니고정대지다당(LPS)유도적소효질세포격활급활화후세포사망적영향. 방법 건립만성니고정폭로적소서동물모형,복강주사LPS유도소효질세포격활,응용면역조직화학방법 관찰피질、해마、흑질CD-11b양성소효질세포표체적변화;BV2세포(소서소효질류세포계)전대배양,운용CCK-8시제합검측세포활성,일양화담검측시제합검측일양화담(NO)석방정황,RT-PCR분석유도형일양화담합매(iNOS)、종류배사인자(TNF-α)、백세포개소1(IL-1β)、백세포개소6(IL-6)、배양화매-2(COX-2)、간우소조절인자1(IRF-1)、Caspase-11 mRNA적표체,면역인적법분석P-I-κB、Caspase-3적표체변화. 결과 니고정억제LPS유도적피질、해마、흑질CD-11b양성소효질세포적표체;니고정억제LPS자격인기적BV2세포적사망,NO적석방,iNOS、TNF-α、IL-1β、IL-6、COX-2、IRF-1、Caspase-11 mRNA적표체,P-I-κB、Caspase-3단백적표체. 결론 니고정가이억제LPS유도적소효질세포활화급격활유도적세포사망(AICD),대뇌내염증반응구유신경보호작용.
Objective To observe the effect of nicotine(NIC) on the activation and resultant death of microglia induced by LPS. Methods The animal model that exposed to chronic nicotine treatment was established and LPS was injected intraperitoneally to induce the activation of microglia. Furthermore, the CD11b-positive microglia in cerebral cortex, hippocampal and substantia ngra were observed through immunohistochemical staining. BV2 cells(Microglial cell line of mouse) were subcultured, simultaneously the following kits were used including CCK-8 kit assay for cell activity, Nitric oxide assay kit assay for NO release, RT-PCR assay for the iNOS,TNF-α,IL-1β,IL-6,COX-2,IRF-1,Caspase-11 mRNA expression, Western blotting assay for the protein expression of P-I-κB and Caspase-3. Results Nicotine suppressed the CD11b-positive microglia expression in cerebral cortex,hippocampal and substantia ngra induced by LPS;Nicotine inhibited the activation-induced cell death (AICD), attenuated NO release, reduced iNOS,TNF-α,IL-1β,IL-6,COX-2,IRF-1,Caspase-11 mRNA expression, decreased the protein expression including P-I-κB and Caspase-3 of BV2 cells. Conclusion Nicotine pretreatment can suppress the activation and resultant death of microglial cells induced by LPS, which suggests that nicotine may play a neuroprotective role on inflammatory reaction of brain.;