CAJ | 학술논문

目的观察Toll样受体9(TLR9)信号通路在大鼠脑梗死灶周围皮质组织中的转导变化. 方法采用线栓法成功制备48只SD大鼠大脑中动脉闭塞(MCAO)模型,并设置假手术组(24只)作为对照.在脑缺血90 min再灌注6h、3d、7d、14d时分别对大鼠进行神经功能缺损评分,取脑组织行TTC染色以测定梗死体积,取梗死灶周围皮质和对侧相应皮质行Western blotting,测定TLR9、核因子NFκB(P65)、肿瘤坏死因子-α(TNFα)、干扰素调节因子-7(IRF7)、干扰素-3(IFNβ)的蛋白表达水平. 结果再灌注6h、3d、7d、14d时,模型组大鼠的神经功能缺损评分和梗死体积随时间的延长逐渐减小；梗死侧TLR9、IRF7、IFNβ蛋白的表达逐渐增加,而NFκB(P65)、TNFα蛋白的表达呈先上升后下降的趋势,表达高峰在7d.同一蛋白的表达量在不同时间点间比较差异均有统计学意义(P＜0.05)；同一观察时间点比较,梗死侧各蛋白的表达量高于对侧相应皮质,差异有统计学意义(P＜0.05).再灌注6h、3d和7d时NFKB(P65)、TNF-α蛋白表达分别高于IRF7、IFNβ,而再灌注14d时NFκB(P65)、TNFα蛋白表达分别低于IRF7、IFNβ,差异均有统计学意义(P＜0.05).在假手术组各时间点均未检测到上述蛋白的表达. 结论大鼠短暂性脑缺血再灌注后,TLR9的炎症通路和细胞保护通路相继被激活,可能参与了脑梗死后炎症损伤和组织修复过程.
목적 관찰Toll양수체9(TLR9)신호통로재대서뇌경사조주위피질조직중적전도변화. 방법 채용선전법성공제비48지SD대서대뇌중동맥폐새(MCAO)모형,병설치가수술조(24지)작위대조.재뇌결혈90 min재관주6h、3d、7d、14d시분별대대서진행신경공능결손평분,취뇌조직행TTC염색이측정경사체적,취경사조주위피질화대측상응피질행Western blotting,측정TLR9、핵인자NFκB(P65)、종류배사인자-α(TNFα)、간우소조절인자-7(IRF7)、간우소-3(IFNβ)적단백표체수평. 결과 재관주6h、3d、7d、14d시,모형조대서적신경공능결손평분화경사체적수시간적연장축점감소；경사측TLR9、IRF7、IFNβ단백적표체축점증가,이NFκB(P65)、TNFα단백적표체정선상승후하강적추세,표체고봉재7d.동일단백적표체량재불동시간점간비교차이균유통계학의의(P＜0.05)；동일관찰시간점비교,경사측각단백적표체량고우대측상응피질,차이유통계학의의(P＜0.05).재관주6h、3d화7d시NFKB(P65)、TNF-α단백표체분별고우IRF7、IFNβ,이재관주14d시NFκB(P65)、TNFα단백표체분별저우IRF7、IFNβ,차이균유통계학의의(P＜0.05).재가수술조각시간점균미검측도상술단백적표체. 결론 대서단잠성뇌결혈재관주후,TLR9적염증통로화세포보호통로상계피격활,가능삼여료뇌경사후염증손상화조직수복과정.
Objective To study the changes of toll-like receptor 9 (TLR9) signaling transduction in the peri-infarct cortex of rats with middle cercbral artery occlusion (MCAO). Methods The MCAO models were established in 48 SD rats using intraluminal filament method and reperfusion was performed by removing the filament 90 min after occlusion.Sham-operated group was established as controls (n=24).At 6 h,and 3,7 and 14 d after reperfusion,National Institutes of Health of neurological deficit scale was performed; the brain tissues were dyed by TTC staining; the cerebral infarct volumes were measured,and the protein expressions of TLR9,nuclear factor-kappaB (NF-kappaB,P65),tumor necrosis factor-α (TNFα),interferon regulatory factor-7 (IRF7) and interferon-β (IFNβ) in the peri-infarct cortex and contralateral cortex were detected by Western blotting. Results Neurological scale scores and infract volumes of the MCAO rats gradually decreased at 6 h,and 3,7 and 14 d after repeffusion;with the time prolongation,the protein expression of TLR9,IRF7 and IFNβ gradually increased,while the protein expressions of NFκ B (P65) and TNFα ascended first and then descended with the highest expression level at 7 d.The differences of the same protein expressions between each 2 time points were statistically significant (P＜0.05). At the same observation time point, the protein expression in the peri-infarct cortex was significantly higher than that in the contralateral cortex (P＜0.05).The expressions of NFκB (P65) and TNF-α were significantly higher than those of IRF7and IFN-β at 6 h,and 3 and 7 d,respectively; but the results were opposite at 14 h (P＜0.05).These proteins were not detected in the sham-operated group at all time points. Conclusions TLR9 inflammatory pathways and cell protection pathways were activated in ischemic cortex, which may be involved in the process of inflammatory impairment and tissue repair after cerebral infarction.