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本研究旨在探讨三七总皂苷(Panax notoginseng saponins,PNS)对大鼠肺缺血/再灌注(ischemia/reperfusion,I/R)损伤时细胞凋亡及凋亡相关蛋白和c-Jun氨基末端激酶(c-Jun N-terminal kinase,JNK)的影响。实验用健康清洁级Wistar大鼠30只,随机分为假手术对照组(Control组)、缺血/再灌注组(I/R组)与缺血/再灌注+三七总皂苷干预组(PNS组)。采用阻断左肺门30min后松开结扎的方法复制在体大鼠原位单肺缺血/再灌注模型,PNS干预方法为缺血前60min与再灌注前10min腹腔注射PNS。实验结束时取部分左肺组织测湿/干重比(wet/dry weight ratio,W/D);采用蛋白免疫印迹法检测肺组织JNK、磷酸化JNK(p-JNK)蛋白的表达;免疫组化法检测肺组织Bcl-2、Bax、Caspase-3蛋白的表达;原位末端标记法(TUNEL)检测肺组织细胞凋亡指数(apoptotic index,AI),光镜、电镜观察肺组织形态结构变化,并测定肺泡损伤数(injured alveolus rate,IAR)。结果显示,与Control组相比,I/R组肺组织p-JNK、Bcl-2、Bax、Caspase-3蛋白的表达均显著增加(均P<0.01),Bcl-2/Bax的比值明显下降(P<0.05),AI、W/D及IAR均显著升高(均P<0.01),光镜、电镜示肺组织结构呈明显损伤性变化;与I/R组相比,PNS组肺组织p-JNK、Bax、Caspase-3蛋白的表达均显著下调(均P<0.01),Bcl-2蛋白的表达及Bcl-2/Bax的比值显著上调(均P<0.01),AI、W/D及IAR也显著降低(均P<0.01),肺组织形态学异常改变不同程度减轻。以上结果提示:PNS对缺血/再灌注肺具有保护作用,该作用可能与其抑制JNK信号转导通路、上调Bcl-2/Bax的比值从而减少Caspase-3依赖性的肺细胞凋亡有关。
This study was designed to investigate the effects of panax notoginseng saponins (PNS) on the apoptosis and the expression of apoptosis-related protein and c-Jun amino terminal in rat lung after ischemia / reperfusion (I / R) injury Effect of c-Jun N-terminal kinase (JNK). Thirty healthy Wistar rats were randomly divided into three groups: control group, ischemia / reperfusion group (I / R group) and ischemia / reperfusion + group). The model of orthotopic lung ischemia / reperfusion in vivo was reproduced by occlusion of the left hilar in 30 minutes and then ligation. The PNS intervention method was intraperitoneal injection of PNS 60 min before ischemia and 10 min before reperfusion. At the end of the experiment, the left / right lung wet weight ratio (W / D) was measured. The expression of JNK and p-JNK protein in lung tissue was detected by Western blotting. Immunohistochemistry The expression of Bcl-2, Bax and Caspase-3 protein in lung tissue was detected by TUNEL method. The apoptotic index (AI) of lung tissue was detected by TUNEL. The morphological changes of lung tissue were observed under light microscope and electron microscope. , And the injured alveolus rate (IAR) was measured. The results showed that the expression of p-JNK, Bcl-2, Bax and Caspase-3 in lung tissue of I / R group were significantly increased (all P <0.01) and the ratio of Bcl-2 / Bax was significantly decreased (P <0.05). The AI, W / D and IAR were significantly increased (all P <0.01). The lung tissue structure was obviously damaged by light microscope and electron microscope. Compared with I / R group, The expressions of Bcl-2 protein and Bcl-2 / Bax were significantly up-regulated (all P <0.01), AI, W / D And IAR were also significantly reduced (all P <0.01), lung tissue morphology changes to varying degrees reduce. The above results suggest that PNS can protect the lung from ischemia / reperfusion injury, which may be related to the inhibition of JNK signal transduction pathway, the up-regulation of Bcl-2 / Bax ratio and the decrease of Caspase-3-dependent apoptosis of lung cells.