Department of Geriatric, Cangzhou Central Hospital, Cangzhou City, Hebei Province, 061001, People's Republic of China.
Department of Anesthesiology, Tianjin Medical University General Hospital, Tianjin Research Institute of Anesthesiology, Tianjin, 300052, People's Republic of China.
Inflammation. 2019 Apr;42(2):426-439. doi: 10.1007/s10753-018-00952-5.
Acute lung injury/acute respiratory distress syndrome (ALI/ARDS) undergoes the process of pathological event including lung tissue dysfunction, pulmonary edema, and inflammation in sepsis. Autophagy is a cytoprotective process recognized as one of the major pathways for degradation and recycling of cellular constituents. Autophagy as a protective or maladaptive response was still confused in ALI during sepsis. Acute lung injury was performed by cecal ligation and puncture (CLP). Autophagic inducer rapacymin and inhibitor 3-MA and autophagosomal-lysosome fusion inhibitor bafilomucin (Baf) A1 and chloroquine (CQ) were administrated by intraperitoneal injection at 1 h after CLP operation. Microtubule-associated protein light chain 3 II (LC3II), Beclin 1, Rab7, and lysosome-associated membrane protein type 2 (LAMP2) were detected by western blotting. Seven-day survival rate of septic mice was observed. Histologic scores, lung wet-to-dry (W/D) weight ratio, oxygenation index (PaO/FiO), total cells and polymorphonuclear neutrophils (PMN) in bronchial alveolar lavage fluid (BALF) and myeloperoxidase (MPO) activity and cytokine tumor necrosis factor (TNF)-α, high-mobility group box (HMGB)1, interleukin (IL)-6, IL-10, and monocyte chemotactic protein (MCP)1 were measured after sham or ALI operation. ALI induced the increasing expression of autophagy-related protein LC3II, Beclin 1, Rab7, and LAMP2 in CLP operation. Autophagic inducer rapacymin significantly induced the expression of LC3II, Beclin 1, LAMP2, and Rab7 in mice model of CLP, and inhibitor 3-MA reduced expression of LC3II, Beclin 1, LAMP2, and Rab7 expressions in CLP + RAP mice compared to CLP group. Compared with ALI group, Baf and CQ obviously elevated the level of LC3II and Beclin 1, and reduced the LAMP2 and Rab7 expressions in CLP + Baf group and ALI + CQ group. Compared with CLP group, autophagic inducer rapacymin improved the survival rate, histologic scores, lung wet/dry weight ratio, PaO/FiO, total cells, and PMNS in BALF and MPO activity and cytokines TNF-α, HMGB1, IL-6, IL-10, and MCP1 in CLP + RAP group, but there were exacerbated above indicators in CLP + 3-MA group, CLP + Baf group, and CLP + CQ group. Autophagy activation participated in the pathophysiologic process of sepsis, and alleviated the cytokine excessive release and lung injury in sepsis.
急性肺损伤/急性呼吸窘迫综合征（ALI/ARDS）在脓毒症中经历包括肺组织功能障碍、肺水肿和炎症在内的病理事件。自噬是一种公认的细胞成分降解和再循环的细胞保护过程，作为主要途径之一。自噬作为一种保护性或适应性反应，在脓毒症中的 ALI 中仍然存在混淆。通过盲肠结扎和穿刺（CLP）来进行急性肺损伤。自噬诱导剂 rapacymin 和抑制剂 3-MA 以及自噬体-溶酶体融合抑制剂巴弗洛霉素 A1（Baf）A1 和氯喹（CQ）在 CLP 手术后 1 小时通过腹腔内注射给药。通过蛋白质印迹法检测微管相关蛋白轻链 3 II（LC3II）、Beclin 1、Rab7 和溶酶体相关膜蛋白 2 型（LAMP2）。观察脓毒症小鼠的 7 天存活率。在假手术或 ALI 手术后测量组织学评分、肺湿重/干重（W/D）比、氧合指数（PaO/FiO）、支气管肺泡灌洗液（BALF）中的总细胞和多形核粒细胞（PMN）以及髓过氧化物酶（MPO）活性和细胞因子肿瘤坏死因子（TNF）-α、高迁移率族蛋白（HMGB）1、白细胞介素（IL）-6、IL-10 和单核细胞趋化蛋白（MCP）1。CLP 手术后，自噬相关蛋白 LC3II、Beclin 1、Rab7 和 LAMP2 的表达增加诱导了 ALI。自噬诱导剂 rapacymin 显著诱导了 CLP 小鼠模型中 LC3II、Beclin 1、LAMP2 和 Rab7 的表达，而抑制剂 3-MA 降低了 CLP+RAP 小鼠中 LC3II、Beclin 1、LAMP2 和 Rab7 的表达与 CLP 组相比。与 ALI 组相比，Baf 和 CQ 明显增加了 CLP+Baf 组和 ALI+CQ 组中 LC3II 和 Beclin 1 的水平，并降低了 LAMP2 和 Rab7 的表达。与 CLP 组相比，自噬诱导剂 rapacymin 提高了存活率、组织学评分、肺湿/干重比、PaO/FiO、BALF 中的总细胞数和 PMNS 以及 MPO 活性和细胞因子 TNF-α、HMGB1、IL-6、IL-10 和 MCP1 在 CLP+RAP 组中，但在 CLP+3-MA 组、CLP+Baf 组和 CLP+CQ 组中上述指标加重。自噬激活参与了脓毒症的病理生理过程，并减轻了脓毒症中细胞因子的过度释放和肺损伤。
