摘要
目的:探讨自噬相关的AMPK/mTOR信号通路对急性肾损伤(acute kidney injury,AKI)诱导的急性肺损伤(acute lung injury,ALI)的影响及作用机制。方法:48只雄性SD大鼠随机分为4组(n=12):假手术组(sham组),缺血-再灌注损伤组(ischemia-reperfusion injury,IRI组),IRI+雷帕霉素组(rapamycin,RA组),IRI+3-甲基腺嘌呤(3-methyladenine,3-MA组)。进行右侧侧腹切口,结扎并切除右肾。左肾经受60 min的缺血,然后再灌注12、24、48和72 h。测定血清肌酐、血尿素氮、肺湿/干比、炎症指标和氧化应激水平,对肺、肾进行病理学检测。通过免疫组织化学和蛋白质印迹法检测AMPK、mTOR、p62、LC3-Ⅱ/LC3-Ⅰ比值、Beclin-1、ULK1的蛋白表达。结果:与sham组大鼠相比,IRI组大鼠在AKI后表现出显著的肺损伤,血清肌酐、血尿素氮、肺湿/干比、炎症指标和氧化应激水平增加(P<0.05)。AMPK、LC3-Ⅱ/LC3-Ⅰ比值、Beclin-1、ULK1的表达增加(P<0.05),而p62和mTOR减少(P<0.01)。此外,RA处理通过激活AMPK/mTOR通路促进自噬显著减轻肺损伤,而3-MA处理通过抑制AMPK/mTOR通路而抑制自噬使肺损伤加重。结论:肾脏IRI后可激活AMPK/mTOR信号通路,并通过该通路诱导自噬,抑制炎症反应、氧化应激和细胞凋亡,从而显著减轻AKI和AKI诱导的ALI。
Objective:To investigate the effects of autophagy-related AMPK/mTOR signaling pathway on acute kidney in-jury(AKI)-induced acute lung injury(ALI)and its mechanism.Methods:The 48 male Sprague-Dawley rats were divided into four groups randomly(n=12):(1)sham,(2)ischemia-reperfusion injury(IRI),(3)IRI+rapamycin(RA),and(4)IRI+3-methyladenine(3-MA).Unilateral flank incision was made and right kidney was ligated and excised.The left kidney was subjec-ted to 60 min of ischemia followed by 12,24,48,and 72 h of reperfusion.The levels of Scr,blood urea nitrogen(BUN),lung of Wet/Dry ratio,indexes of inflammation,and oxidative stress were assayed.Histological examinations of lung and kidney were per-formed.The protein expression of AMPK,mTOR,p62,LC3-II/LC3-I ratio,and Beclin-1,ULK1 was evaluated by western blotting and immunohistochemistry.Results:Compared to the rats from the sham group,IRI rats showed significantly pulmonary dam-age after AKI with increased Scr,BUN,lung of Wet/Dry ratio,indexes of inflammation,and oxidative stress(P<0.05).The ex-pression of AMPK,LC3-I/LC3-I ratio,Beclin-1,and ULK1 and were increased(P<0.05),while p62 and mTOR were de-creased(P<0.01).In addition,RA treatment significantly attenuated lung injury by promoting autophagy through the activation of the AMPK/mTOR signaling pathway,and 3-MA treatment exhibited adverse effects inversely.Conclusion:Renal IRI could activate AMPK/mTOR signaling pathway and induce autophagy through this pathway,inhibit inflammatory response,oxidative stress and ap-optosis,thereby significantly alienate AKI and AKI-induced ALI.
作者
简禄勇
袁琦
曹海虹
张星炜
黄仁发
JIAN Luyong;YUAN Qi;CAO Haihong(Department of Nephropathy,Guangzhou University of Chinese Medicine Sixth Clinical Medical College,Shenzhen518034)
出处
《中国中西医结合肾病杂志》
2024年第5期395-402,407,I0004-I0007,共13页
Chinese Journal of Integrated Traditional and Western Nephrology
基金
广东省自然科学基金资助项目(No.2020A1515010566)
深圳市科技研发基金资助项目(No.JCYJ20190809102413156)。
