Wu Yu, Wang Ling, Meng Lei, Cao Guang-Ke, Zhao Yu-Liang, Zhang Yang
Department of Nephrology, Xuzhou Municipal Hospital Affiliated to Xuzhou Medical University, Xuzhou, Jiangsu 221000, P.R. China.
Department of Intensive Care Unit, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, Jiangsu 221000, P.R. China.
Exp Ther Med. 2019 Jan;17(1):316-322. doi: 10.3892/etm.2018.6899. Epub 2018 Oct 30.
This study investigated whether autophagy is activated after sepsis-induced acute kidney injury (AKI) and explored its biological role. Seventy-two normal C57 mice were randomly divided into sham operation group, cecal ligation and puncture (CLP) group and CLP+3-MA (autophagy inhibitor) group; 24 mice in each group. Mice in CLP and CLP+3-MA group were treated with cecal ligation to establish sepsis, while mice in sham operation group were treated with the same surgical operations, but not cecal ligation. Blood samples were collected from 12 mice of each group and the levels of serum creatinine (Cr) and blood urea nitrogen (BUN) were measured. The pathological changes were observed. The remaining 12 mice in each group were kept and the survival rate was recorded. Changes in the expressions of autophagy-related proteins were detected by reverse transcription-semi-quantitative PCR and western blotting. The results revealed that the levels of Cr and BUN in CLP and CLP+3-MA group were significantly higher than those in sham operation group (P<0.05), and the levels of Cr and BUN in CLP+3-MA group were higher than those in CLP group (P<0.05). The pathological score of renal injury in CLP+3-MA group was significantly higher than that of CLP group (P<0.01). The expression levels of Beclin1 and LC3-II/I were significantly increased in CLP group compared to sham operation group (P<0.01), while the expression of p62 was decreased (P<0.01). After 3-MA treatment the expression levels of Beclin1 and LC3-II/I were decreased, compared with CLP group, but accumulation of p62 occurred, and the degree of renal injury was increased. In conclusion, AKI induced by sepsis in mice can induce apoptosis and activate autophagy. The activation of autophagy aggravates the renal injury in mice, which in turn inhibits AKI after sepsis.
本研究调查了脓毒症诱导的急性肾损伤（AKI）后自噬是否被激活，并探讨了其生物学作用。将72只正常C57小鼠随机分为假手术组、盲肠结扎穿孔（CLP）组和CLP + 3 - MA（自噬抑制剂）组；每组24只小鼠。CLP组和CLP + 3 - MA组小鼠行盲肠结扎以建立脓毒症模型，而假手术组小鼠接受相同的外科手术操作，但不行盲肠结扎。从每组12只小鼠采集血样，检测血清肌酐（Cr）和血尿素氮（BUN）水平。观察病理变化。每组剩余12只小鼠饲养并记录存活率。通过逆转录 - 半定量PCR和蛋白质印迹法检测自噬相关蛋白表达的变化。结果显示，CLP组和CLP + 3 - MA组的Cr和BUN水平显著高于假手术组（P < 0.05），且CLP + 3 - MA组的Cr和BUN水平高于CLP组（P < 0.05）。CLP + 3 - MA组肾损伤病理评分显著高于CLP组（P < 0.01）。与假手术组相比，CLP组中Beclin1和LC3 - II/I的表达水平显著升高（P < 0.01），而p62表达降低（P < 0.01）。3 - MA处理后，与CLP组相比，Beclin1和LC3 - II/I的表达水平降低，但p62出现蓄积，且肾损伤程度增加。总之，小鼠脓毒症诱导的AKI可诱导凋亡并激活自噬。自噬的激活加重了小鼠的肾损伤，进而抑制脓毒症后的AKI。
