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目的研究姜黄素(curcumin,CUR)对肺缺血/再灌注(ischemia/reperfusion,I/R)损伤小鼠天冬氨酸特异性半胱氨酸蛋白酶-12(cysteinyl aspartate specific proteinase-12,Caspase-12)及细胞凋亡的干预作用。方法采用C57BL/6J小鼠制作在体单侧肺原位I/R损伤模型。采用随机数字表法将60只小鼠分为6组,每组10只。假手术组(简称Sham组)、I/R组、I/R加二甲基亚砜(简称I/R加DMSO)组,I/R分别加100、150、200 mg/kg CUR(简称I/R加CUR-100、I/R加CUR-150、I/R加CUR-200)组。实验结束留取左肺,测定肺湿/干重比(W/D)和总肺水含量(TLW),光镜观察肺组织形态学改变,进行肺组织损伤定量评估(IQA),电镜观察肺组织超微结构改变,逆转录-聚合酶链反应(RT-PCR)和蛋白免疫印迹法(Western blot)分别检测Caspase-12和葡萄糖调节蛋白78(GRP78)mRNA及蛋白表达水平,原位末端标记(TUNEL)法检测肺组织细胞凋亡指数(AI)。结果与Sham组比较,I/R组Caspase-12和GRP78 mRNA及蛋白表达水平均升高(P<0.05),W/D、TLW、IQA和AI均增加(P<0.05,P<0.01),肺组织形态结构均发生明显损伤;与I/R加DMSO组比较,I/R加CUR-1 00组、I/R加CUR-1 50组、I/R加CUR-200组GRP78 mRNA及蛋白表达水平均升高(P<0.05),而Caspase-12 mRNA及蛋白表达水平均降低(P<0.05),W/D、TLW、IQA和AI亦均下降(P<0.05,P<0.01),肺组织形态学异常改变趋近于正常。结论姜黄素对I/R损伤发生的小鼠肺脏具有较好的保护作用,其机制可能与其对抗过度的未折叠蛋白反应(unfolded protein response,UPR)中Caspase-12引起的细胞凋亡有关。
Objective To investigate the effect of curcumin (CUR) on the expression of cysteinyl aspartate specific proteinase-12 (Caspase) in mice with lung ischemia / reperfusion (I / R) -12) and apoptosis. Methods C57BL / 6J mice were used to establish I / R injury model in situ in unilateral lung. Sixty mice were divided into 6 groups using random number table method, 10 in each group. Sham group, I / R group, I / R plus dimethylsulfoxide (I / R plus DMSO for short), I / R were added 100,150,200 mg / kg CUR / R plus CUR-100, I / R plus CUR-150, I / R plus CUR-200). At the end of the experiment, left lungs were taken for determination of wet / dry weight ratio (W / D) and total lung water content (TLW). Lung morphological changes were observed under light microscope. Quantitative assessment of lung injury (IQA) Tissue ultrastructural changes, RT-PCR and Western blot were used to detect the expression of Caspase-12 and GRP78 mRNA and protein, respectively. The in situ terminal markers (TUNEL) method was used to detect the apoptosis index (AI) of lung tissue. Results Compared with Sham group, the mRNA and protein expressions of Caspase-12 and GRP78 in I / R group were increased (P <0.05), W / D, TLW, IQA and AI were increased Compared with the I / R group plus the CUR-100 group, I / R plus the CUR-1 50 group, the I / R plus the CUR-200 group, the GRP78 mRNA and protein (P <0.05), while the expression of Caspase-12 mRNA and protein were decreased (P <0.05), W / D, TLW, IQA and AI also decreased Abnormal changes in lung morphology approached normal. CONCLUSION: Curcumin has a good protective effect on the lungs of mice with I / R injury. The mechanism may be related to the apoptosis induced by caspase-12 in the unfolded protein response (UPR).