骨髓间充质干细胞通过抑制铁死亡改善大鼠重症急性胰腺炎相关肺损伤  

Bone marrow-derived mesenchymal stem cells ameliorate severe acute pancreatitis-induced lung injury in rats via suppression of ferroptosis

作　　者：余伟迪 宋振顺 YU Weidi;SONG Zhenshun(Department of General Surgery,Shanghai Tenth People's Hospital,Clinical College of Nanjing Medical University,Shanghai 200072,China;Department of General Surgery,Shanghai Tenth People's Hospital,Tongji University School of Medicine,Shanghai 200072,China)

机构地区：[1]南京医科大学上海十院临床医学院,上海200072 [2]同济大学附属上海市第十人民医院肝外科,上海200072

出　　处：《中国临床药理学与治疗学》2022年第7期721-728,共8页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：国家自然科学基金项目(81670582)。

摘　　要：目的:探讨骨髓间充质干细胞(BMSCs)对重症急性胰腺炎(SAP)大鼠肺组织内细胞铁死亡的抑制作用及其潜在机制。方法:选择3周龄SD雄性大鼠提取、培养BMSCs。将30只6周龄SD雄性大鼠随机分为5组:假手术组(Sham)、胰腺炎组(SAP)、PBS对照组(SAP+PBS)、干细胞治疗组(SAP+BMSCs)、干细胞联合Nrf2阻断剂ML385治疗组(SAP+BMSCs+ML385),每组6只。使用牛磺胆酸钠(NaT)构建大鼠SAP模型。于造模前2 h,SAP+BMSCs+ML385组腹腔注射ML385(5 mg/kg);于造模后6 h,SAP+BMSCs组和SAP+BMSCs+ML385组尾静脉注射BMSCs(1×10^(7)/只),SAP+PBS组尾静脉注射等量的磷酸盐缓冲液(PBS)。3 d后处死大鼠,取胰腺、肺组织和血清。H&E染色观察组织损伤情况。检测血清中淀粉酶、脂肪酶活性和炎症因子水平,肺组织中铁含量、丙二醛(MDA)、谷胱甘肽(GSH)活性。检测肺组织铁响应元件结合蛋白2(IREB2)、铁蛋白重链1(FTH1)、长链脂酰CoA合成酶4(ACSL4)、谷胱甘肽过氧化物酶4(GPX4)、溶质载体家族7成员11(SLC7A11)等铁死亡相关蛋白表达,以及PI3K/AKT/Nrf2通路相关蛋白表达。结果:NaT诱导后大鼠血清淀粉酶、脂肪酶活性、促炎因子水平和病理评分明显升高(P<0.05)。BMSCs移植能够明显减轻上述指标(P<0.05)。相比SAP组,BMSCs治疗组肺组织铁、MDA水平降低,GSH活性升高,IREB2、ACSL4蛋白表达降低,FTH1、GPX4及SLC7A11蛋白表达升高(P<0.05);ML385能够部分抵消BMSCs移植产生的抗脂质过氧化作用(P<0.05)。BMSCs移植上调了肺组织PI3K、p-AKT总蛋白和Nrf2核蛋白的表达(P<0.05)。结论:BMSCs移植能够有效减轻SAP相关肺损伤的全身炎症反应和局部组织损伤,抑制肺组织铁沉积和脂质过氧化,其抗脂质过氧化作用可能与PI3K/AKT/Nrf2通路有关。AIM:To evaluate the effect of bone marrow-derived mesenchymal stem cells(BMSCs)on ferroptosis in lung injury caused by severe acute pancreatitis(SAP).METHODS:BMSCs were obtained from male SD rats(3 weeks of age).30 male SD rats(6 weeks of age)were randomly divided into 5 groups:Sham group,SAP group,SAP+PBS group,SAP+BMSCs group,and SAP+BMSCs+ML385 group,with 6 rats in each group.SAP model induced by sodium taurocholate(NaT)was established.Rats in the SAP+MSCs+ML385 group were intraperitoneally injected with ML385(5 mg/kg)2 h before SAP induction.For PBS control and BMSCs treatment groups,PBS and BMSCs were respectively injected into rats via the tail vein 6 h after creation of the SAP model.All rats were sacrificed 3 days after SAP induction,and pancreas and lung tissue and serum samples were collected.Pancreatic and pulmonary injury was assessed using H&E staining.Serum levels of amylase and lipase and inflammatory factors,as well as tissue levels of iron,malondialdehyde(MDA),glutathione(GSH)were evaluated.The expressions of ferroptosis-associated proteins,including iron responsive element binding protein 2(IREB2),ferritin heavy chain 1(FTH1),acyl-CoA synthetase long-chain family member 4(ACSL4),glutathione peroxidase 4(GPX4),and solute carrier family 7 member 11(SLC7 A11),as well as the expressions of PI3 K/AKT/Nrf2 pathway-related proteins were assessed.RESULTS:The serum levels of amylase,lipase and proinflammatory factors,as well as pathological scores were significantly increased after SAP induction(P<0.05).Changes above were effectively alleviated by BMSCs(P<0.05).Compared with SAP group,the iron and MDA contents were decreased,the GSH activity was increased,the expression of IREB2 and ACSL4 were downregulated and the expression of FTH1,GPX4 and SLC7 A11 were upregulated in SAP+BMSCs group(P<0.05).Injection of ML385 partially offset the anti-lipid peroxidation effect of BMSCs(P<0.05).In addition,BMSCs upregulated the total protein levels of PI3 K and p-AKT,and the nuclear protein level of Nrf2 in lung tissu

关 键 词：间充质干细胞 重症急性胰腺炎 肺损伤 铁死亡 

分 类 号：R657.5[医药卫生—外科学]