自噬抑制细胞焦亡对脓毒症肺损伤的保护作用
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摘要
为研究自噬抑制细胞焦亡对脓毒症肺损伤的保护作用,研究先于体外使用盲肠内容物刺激小鼠腹腔巨噬细胞使其发生炎性反应,通过乳酸脱氢酶检测法、酶联免疫吸附(ELISA)法及Western blottin法检测雷帕霉素激活自噬对巨噬细胞发生细胞焦亡的影响。结果发现激活自噬能降低巨噬细胞炎性小体的活化及炎性细胞因子的释放,抑制细胞焦亡。随后对健康C57小鼠构建脓毒症模型,观察雷帕霉素预处理对各组小鼠死亡率的影响,并通过Western blotting检测肺组织中LC3蛋白表达变化,HE染色观察肺组织形态学改变,发现雷帕霉素预处理能够减轻脓毒症小鼠肺组织病理损伤,降低小鼠死亡率。结果表明自噬能够抑制细胞焦亡,对降低脓毒症急性肺损伤具有重要意义。
Peritoneal macrophages from C57 mice were stimulated by the cecal contents to induce inflammatory response for investigating the effect of autophagy on inhibiting cell pyroptosis and protecting mice from septic lung injury. Lactate dehydrogenase(LDH) assay,enzyme linked immunosorbent assay(ELISA) and Western blotting were used to detect the effect of rapamycin activated autophagy on the occurrence of pyroptosis of macrophages. We found that rapamycin induced autophagy decreased the activation of inflammasomes and the release of inflammatory cytokine from peritoneal macrophages and inhibited pyroptosis. Then,healthy C57 mice were given the operation of cecum ligation and puncture(CLP). Mortality of mice in each group was recorded. Western blotting was used to detect the expression of LC3 in lung samples. Morphological changes of the lung were investigated by HE staining. Results showed that autophagy alleviated the pathological damage of lung tissue and reduced the mortality of septic mice. The results indicated that autophagy can inhibit pyroptosis and reduce acute lung injury induced by sepsis.
Peritoneal macrophages from C57 mice were stimulated by the cecal contents to induce inflammatory response for investigating the effect of autophagy on inhibiting cell pyroptosis and protecting mice from septic lung injury. Lactate dehydrogenase(LDH) assay,enzyme linked immunosorbent assay(ELISA) and Western blotting were used to detect the effect of rapamycin activated autophagy on the occurrence of pyroptosis of macrophages. We found that rapamycin induced autophagy decreased the activation of inflammasomes and the release of inflammatory cytokine from peritoneal macrophages and inhibited pyroptosis. Then,healthy C57 mice were given the operation of cecum ligation and puncture(CLP). Mortality of mice in each group was recorded. Western blotting was used to detect the expression of LC3 in lung samples. Morphological changes of the lung were investigated by HE staining. Results showed that autophagy alleviated the pathological damage of lung tissue and reduced the mortality of septic mice. The results indicated that autophagy can inhibit pyroptosis and reduce acute lung injury induced by sepsis.
引文
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[12] Labbé K,Saleh M.Cell death in the host response to infection [J].Cell Death & Differentiation,2008,15(9):1339-49.
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[18] Kim K H,Jeong Y T,Oh H,et al.Autophagy deficiency leads to protection from obesity and insulin resistance by inducing Fgf21 as a mitokine [J].Nature Medicine,2013,19(1):83-92.
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[20] 韩靖,杜玉明,袁聪俐,等.大鼠盲肠结扎与穿刺脓毒症模型的建立与分析[J].上海交通大学学报(农业科学版),2017(4):1-6.