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目的:探讨丹酚酸B对急性脑缺血小鼠的抗炎作用及Toll样受体4/核转录因子-κB(TLR4/NF-κB)信号通路的影响。方法:建立小鼠大脑中动脉阻塞(MCAO)模型。造模后阳性组(尼莫地平30μg/kg)、丹酚酸B低剂量组(丹酚酸B 11.25 mg/kg)、丹酚酸B高剂量组(丹酚酸B 22.5 mg/kg)分别尾静脉注射给药1次,模型组和假手术组给予同等体积的生理盐水。造模后6 h,分别采用酶联免疫吸附法测定脑组织中IL-1β及TNF-α的含量,采用实时定量PCR法检测缺血侧脑皮层TLR4、NF-κBp65 mRNA的表达,采用Western blot法检测缺血侧脑皮层TLR4、NF-κBp65蛋白的表达。结果:与模型组比较,丹酚酸B高、低剂量组均能显著降低小鼠脑组织中IL-1β、TNF-α含量(P<0.01,P<0.05);丹酚酸B高、低剂量组小鼠缺血侧脑皮层TLR4 mRNA、NF-κBp65 mRNA表达显著降低(P<0.01,P<0.05);丹酚酸B高剂量组小鼠缺血侧脑皮层TLR4、NF-κB蛋白表达显著降低(P<0.01)。结论:丹酚酸B可通过抑制TLR4/NF-κB信号通路,降低TLR4和NF-κB相关基因蛋白的表达,减少炎症因子IL-lβ和TNF-α的含量,进而发挥抗脑缺血的作用。
Objective: To investigate the anti-inflammatory effect of salvianolic acid B on acute cerebral ischemia mice and the effect of Toll-like receptor 4 / TLR4 / NF-κB signaling pathway. Methods: The mouse model of middle cerebral artery occlusion (MCAO) was established. Positive model group (nimodipine 30μg / kg), salvianolic acid B low dose group (salvianolic acid B 11.25 mg / kg) and salvianolic acid B high dose group (salvianolic acid B 22.5 mg / kg) The tail vein injection administered once, the model group and sham operation group given the same volume of saline. The levels of IL-1β and TNF-α in brain tissue were determined by enzyme-linked immunosorbent assay (ELISA) 6 h after model establishment. The expression of TLR4 and NF-κBp65 mRNA in the ischemic cortex were detected by real-time quantitative PCR. Western blot Method to detect the expression of TLR4 and NF-κBp65 protein in ischemic cortex. Results: Compared with the model group, the levels of IL-1β and TNF-α in the brain of the salvianolic acid B high and low dose groups were significantly decreased (P <0.01, P <0.05) The expression of TLR4 mRNA and NF-κBp65 mRNA in ischemic cortex decreased significantly in the dose group (P <0.01, P <0.05), while the expression of TLR4 and NF-κB in the ischemic cortex of the high dose salvianolic acid group Significantly lower (P <0.01). CONCLUSION: Salvianolic acid B can inhibit the expression of TLR4 and NF-κB gene and decrease the levels of inflammatory cytokines IL-1β and TNF-α by inhibiting TLR4 / NF-κB signaling pathway and then exert anti-cerebral ischemia .