κ-阿片受体激动剂对体外循环大鼠肺细胞焦亡和NLRP3炎症小体的影响  

Effects ofκ-opioid receptor agonist U50448H on pyroptosis and NLRP3 of lung in rats with cardiopulmonary bypass

作　　者：李龙[1] 高光洁 Li Long;Gao Guangjie(Department of Anesthesiology,Air Force Hospital of Northern Theater Command of Chinese PLA,Liaoning Shenyang 110042,China)

机构地区：[1]中国人民解放军北部战区空军医院麻醉科,沈阳市110042

出　　处：《中国体外循环杂志》2024年第4期308-312,共5页Chinese Journal of Extracorporeal Circulation

基　　金：辽宁省自然基金资助项目(2020-MS-043)。

摘　　要：目的通过观察κ-阿片受体(KOR)激动剂U50488H对心肺转流(CPB)大鼠肺组织NLRP3炎症小体和细胞焦亡的影响,探讨U50488H减轻CPB致急性肺损伤(ALI)的可能机制。方法24只成年雄性清洁级SD大鼠,350~450 g,随机分为假手术组(Sham组)、CPB组和CPB+KOR激动剂组(U50448H组),每组8只。U50448H组于CPB前30 min静脉注射1.5 mg/kg U50488H。分别于CPB后0 h、1 h和2 h各时点行动脉血气分析,计算肺泡-动脉氧分压差(AaDO2)和呼吸指数(RI)。三组大鼠均在停CPB后2 h时处死,取完整右肺下叶,采用重力法测定血管外肺水(EVLW),HE染色观察肺组织形态学变化。采用ELISA方法检测血浆脂多糖(LPS)、肿瘤坏死因子-α(TNF-α)、白介素(IL)-6和IL-4的含量,Western blot测定肺组织GSDMD-C、GSDMD-N和NLRP3、半胱氨酸天冬氨酸酶前体(pro-Caspase-1)蛋白的表达。结果CPB组大鼠在CPB后0 h、1 h和2 h各时点的AaDO2和RI、LPS明显高于Sham组(P<0.05);U50488组三个时点的AaDO2和RI、LPS明显低于CPB组(P<0.05)。与Sham组比较,CPB组大鼠EVLW、血浆TNF-a和IL-6、肺组织GSDMD-C、GSDMD-N和NLRP3、pro-Caspase-1表达明显增高,血浆IL-4水平明显降低,其差异有统计学意义(P<0.05);而U50488组上述指标与Sham组无明显差异(P>0.05)。CPB组大鼠出现了严重的肺损伤和肺泡内充血/出血,并伴有广泛的炎症细胞浸润,U50448H组肺损伤明显减轻。结论KOR激动剂U50488H对CPB所致ALI期间NLRP3炎性小体诱导的细胞焦亡具有抑制作用,从而减轻了CPB后的肺损伤。Objective To explore the potential protective mechanism ofκ-opioid receptor agonist U50488H in acute lung injury(ALI)induced by cardiopulmonary bypass(CPB)through observing its effects on NLRP3 inflammasome and pyroptosis in lung tissues of rats undergoing CPB.Methods 24 adult male SD rats weighing 350-450 g,were randomly divided into three groups:sham operation group(sham group),CPB group,CPB+KORs agonist group(U50488H group).U50488H 1.5 mg/kg was intravenously injected 30 minutes before CPB in group U50488H.Arterial blood gas analysis was performed at 0 h,1 h,and 2 h after CPB,and the alveolar-arterial oxygen difference(AaDO2)and respiratory index(RI)were calculated.All rats in the three groups were sacrificed 2 hours after wean off CPB.The intact right lower lobe of the lung was taken,and the extravascular lung water(EVLW)was measured by the gravimetric method.HE staining was used to observe the morphological changes of lung tissue.ELISA was applied to test the level of LPS,TNF-α,IL-6 and IL-4 in serum.Western blot was performed for examining the expression of GSDMD-C,GSDMD-N,NLRP3 and pro-Caspase-1 in lung tissue.Results The values of AaDO2,RI and LPS at 0 h,1 h and 2 h after CPB in CPB group were higher than those in sham group(P<0.05),while the values of AaDO2,RI and LPS in U50488H group were significantly lower than those in CPB group(P<0.05).Compared with the sham group,the levels of EVLW,plasma TNF-α,IL-6 and lung GSDMD-C,GSDMD-N,NLRP3,pro-Caspase-1 were increased,and the levels of plasma IL-4 was decreased in CPB group,which were statistically significant(P<0.05),while the above indexes were not significantly different in U50488H group(P>0.05).The rats of CPB group performed serious lung damage,intra-alveolar congestion/hemorrhage and massive inflammatory cell infiltration.The lung injury was alleviated in U50448H group.Conclusion The KOR agonist U50488H inhibits NLRP3 inflammasome-induced pyroptosis during ALI induced by CPB,thereby mitigating post-CPB lung injury.

关 键 词：κ-阿片受体激动剂 心肺转流 急性肺损伤 NLRP3炎症小体 细胞焦亡 

分 类 号：R654.1[医药卫生—外科学]