Objective: To detect the expression of Toll-like receptor 4(TLR-4) and NF-κB and to discuss the mechanism of TLR-4/NF-κB pathway in the myocardial ischemia reperfusion injury of mouse. Methods: TLR-4 mutant mice and...Objective: To detect the expression of Toll-like receptor 4(TLR-4) and NF-κB and to discuss the mechanism of TLR-4/NF-κB pathway in the myocardial ischemia reperfusion injury of mouse. Methods: TLR-4 mutant mice and wild homozygous mice were divided into the model group and sham group. Mice in the model group were given the ligation of left anterior descending coronary artery for the modeling, while mice in the sham group were not given the ligation after threading. The cardiac muscle tissues were collected for the morphological observation. The immuno histochemistry was employed to detect the expression of NF-κB, Western blot was used to detect the expression of TLR-4 and ELISA to detect the expression of serum inflammatory factors. Results: The expression of NF-κB in TLR-4 null mice after the myocardial ischemia reperfusion was significantly lower than that in wild homozygous mice. For the model group and sham group, the expression of TLR-4 in wild homozygous mice was all significantly higher than that in TLR-4 null mice, while the expression of TLR-4 in TLR-4 null mice in the model group was significantly higher than that in sham group, with the statistical difference(P<0.05). The expression of inflammatory factors in TLR-4 null mice and wild homozygous mice in the model group was significantly higher than that in sham group. The expression of all factors in group A with TLR-4 null was significantly lower than that in group B with wild homozygous type, with the statistical difference(P<0.05). Conclusions: TLR-4/NF-κB pathway is closely related to the myocardial ischemia reperfusion injury, which plays its role through the release of inflammatory cytokines.展开更多
文摘目的探讨加味丹参饮预处理是否通过调节Beclin-1和Atg5表达调控自噬抗缺血再灌注损伤大鼠心肌。方法将60只健康SD大鼠随机分为空白对照(control group,C)组、假手术(sham,S)组、缺血再灌注损伤(ischemia reperfusion injury,IRI)组、IRI+加味丹参饮(Jiawei Danshen Yin,JDY)组、IRI+JDY+自噬抑制剂(inhibitor,I)组,每组12只。通过结扎-放松大鼠左冠状动脉前降支制备心肌IRI模型。通过氯化三苯基四氮唑(TTC)染色观察心肌梗死面积率;在透射电镜下观察自噬泡;采用实时荧光定量逆转录-聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)方法检测心肌Beclin-1和Atg5 m RNA表达;采用蛋白质印迹(western blot)法检测心肌Beclin-1蛋白表达变化。结果 IRI+JDY组心肌梗死面积率显著低于IRI组及IRI+JDY+I组(P<0.01)。电镜结果显示,IRI+JDY组适度调节大鼠缺血再灌注损伤心肌细胞的自噬,改善心肌细胞结构。IRI组大鼠心肌细胞中Beclin-1和Atg5 m RNA表达水平及Beclin-1蛋白表达较SG组显著升高(P<0.01);IRI+JDY组、IRI+JDY+I组大鼠心肌细胞中Beclin-1和Atg5 m RNA表达水平及Beclin-1蛋白表达较IRI组显著降低(P<0.01);IRI+JDY组大鼠心肌细胞中Beclin-1和Atg5 m RNA表达水平及Beclin-1蛋白表达较IRI+JDY+I组显著升高(P<0.01)。结论加味丹参饮通过调节缺血再灌注损伤大鼠心肌细胞自噬相关基因Beclin-1和Atg5表达适度,调控缺血再灌注心肌细胞发生适度自噬,从而发挥细胞保护作用。
基金supported by Research Fund of Health Bureau of Hebei Province(Project No.20090601)
文摘Objective: To detect the expression of Toll-like receptor 4(TLR-4) and NF-κB and to discuss the mechanism of TLR-4/NF-κB pathway in the myocardial ischemia reperfusion injury of mouse. Methods: TLR-4 mutant mice and wild homozygous mice were divided into the model group and sham group. Mice in the model group were given the ligation of left anterior descending coronary artery for the modeling, while mice in the sham group were not given the ligation after threading. The cardiac muscle tissues were collected for the morphological observation. The immuno histochemistry was employed to detect the expression of NF-κB, Western blot was used to detect the expression of TLR-4 and ELISA to detect the expression of serum inflammatory factors. Results: The expression of NF-κB in TLR-4 null mice after the myocardial ischemia reperfusion was significantly lower than that in wild homozygous mice. For the model group and sham group, the expression of TLR-4 in wild homozygous mice was all significantly higher than that in TLR-4 null mice, while the expression of TLR-4 in TLR-4 null mice in the model group was significantly higher than that in sham group, with the statistical difference(P<0.05). The expression of inflammatory factors in TLR-4 null mice and wild homozygous mice in the model group was significantly higher than that in sham group. The expression of all factors in group A with TLR-4 null was significantly lower than that in group B with wild homozygous type, with the statistical difference(P<0.05). Conclusions: TLR-4/NF-κB pathway is closely related to the myocardial ischemia reperfusion injury, which plays its role through the release of inflammatory cytokines.
