低氧预处理胎盘间充质干细胞减轻重症急性胰腺炎模型小鼠胰腺组织损伤

doi:10.16352/j.issn.1001-6325.2023.09.1346

基础医学与临床 ›› 2023, Vol. 43 ›› Issue (9): 1346-1352.doi: 10.16352/j.issn.1001-6325.2023.09.1346

低氧预处理胎盘间充质干细胞减轻重症急性胰腺炎模型小鼠胰腺组织损伤

景光旭^1,2, 王张鹏^1,2, 刘忠钰^1,2, 吕双^2,3, 孙红玉^1,2,3*

1.西南医科大学临床医学院肝胆外科,四川泸州 646000;
2.中国人民解放军西部战区总医院四川省胰腺损伤与修复重点实验室,四川成都 610083;
3.西南交通大学医学院, 四川成都 610063

收稿日期:2022-12-12 修回日期:2023-04-14 出版日期:2023-09-05 发布日期:2023-09-01
通讯作者: ^*shongyu2008@163.com
基金资助:
国家自然科学基金(81772001)

Hypoxic preconditioning placental mesenchymal stem cells alleviate pathological lesion in mouse models with severe acute pancreatitis

JING Guangxu^1,2, WANG Zhangpeng^1,2, LIU Zhongyu^1,2, LYU Shuang^2,3, SUN Hongyu^1,2,3*

1. Department of Hepatobiliary Surgery, School of Clinical Medicine, Southwest Medical University, Luzhou 646000;
2. Sichuan Provincial Key Laboratory of Pancreatic Injury and Repair, the Chinese PLA Western Theater Command General Hospital,Chengdu 610083;
3. College of Medicine, Southwest Jiaotong University, Chengdu 610063, China

Received:2022-12-12 Revised:2023-04-14 Online:2023-09-05 Published:2023-09-01
Contact: ^*shongyu2008@163.com

摘要/Abstract

摘要： 目的探讨低氧预处理胎盘间充质干细胞(HP-MSCs)对小鼠重症急性胰腺炎(SAP)组织损伤的保护作用,并探索其可能机制。方法分别于常氧(21% O₂)和低氧(1% O₂)培养48 h的胎盘间充质干细胞(P-MSCs),获得常压和低氧预处理胎盘间充质干细胞(N-MSCs和HP-MSCs),CCK-8和MTT法检测P-MSCs活力与迁移能力;C57BL/6雄性小鼠32只,分为假手术组(sham)、SAP组(胰胆管逆行注射4%牛磺胆酸钠0.1 mL/100 g)、常氧组(N-MSCs)和低氧组(HP-MSCs)。造模结束后6 h,经尾静脉注射N-MSCs和HP-MSCs(1×10⁶个/100 g)干预SAP,每组8只。在造模后12 h收集血清和胰腺组织。HE染色评估胰腺组织损伤程度并进行病理评分;ELISA测定血清淀粉酶、脂肪酶活性及炎性因子水平;Western blot检测受体相互作用蛋白3(RIP3)、MLKL、MLKL的磷酸化含量(p-MLKL)、NLRP3在胰腺组织中的表达。结果 HP-MSCs在体外表现出较N-MSCs更好的活力、增殖能力;在小鼠体内,与SAP组相比,N-MSCs组中胰腺病理评分降低(P＜0.05),血清中淀粉酶、脂肪酶活性降低,IL-6、 TNF-α水平降低(P＜0.05);与N-MSCs组相比,HP-MSCs组中胰腺病理评分降低(P＜0.05),炎性因子TNF-α、IL-6水平降低(P＜0.05)。在HP-MSCs中,RIP3、p-MLKL、NLRP3在胰腺组织的表达中明显低于SAP组与N-MSCs组(P＜0.05)。结论 HP-MSCs 较N-MSCs更能显著减轻 SAP,其机制可能与抑制RIP1/RIP3-MLKL信号通路发挥其作用有关。

关键词: 低氧预处理, 胎盘间充质干细胞, 重症急性胰腺炎, 受体相互作用蛋白3(RIP3)

Abstract: Objective To investigate the protective effect of hypoxia-preconditioned placental mesenchymal stem cells (HP-MSCs) on severe acute pancreatitis (SAP) of mice model and to explore its mechanisms. Methods Placental-derived mesenchymal stem cells (P-MSCs) were cultured in normoxia treatment(N-MSCs)(21% O₂) or hypoxia preconditioning(HP-MSCs)(1% O₂) for 48 h. CCK-8 and MTT assay were used to detect the viability andmigration of N-MSCs and HP-MSCs respectively. Thirty-two male C57BL/6 mice were divided into sham operation group (sham), SAP group (retrograde injection of 4% sodium taurocholate 0.1 mL/100 g into the pancreaticobiliary duct), normoxia group (N-MSCs) and hypoxia group (HP-MSCs). Six hours after the end of modeling, N-MSCs and HP-MSCs (1×10⁶/100 g) were injected to treat SAP, with 8 rats in each group. Serum and pancreatic tissue were collected at 12 h after modeling. HE staining and microscopy were used to evaluate the degree of pancreatic tissue injury and pathological score. ELISA was used to measure the activities of serum amylase and lipase and the levels of inflammatory factors. Western blot was used to detect the expression of receptor interacting protein 3 (RIP3), MLKL, phosphorylated MLKL (p-MLKL) and NLRP3 in pancreatic tissue. Results HP-MSCs showed better viability and higher capacity proliferation than N-MSCs in vitro. In vivo, compared with SAP group, the pancreatic pathological score was significantly decreased in N-MSCs group (P＜0.05). The biological activity of amylase and lipase in serum were decreased, and the level of IL-6 and TNF-α was decreased (P＜0.05). Compared with N-MSCs group, the pancreatic pathological score and the level of TNF-α and IL-6 in HP-MSCs group were significantly decreased(P＜0.05). The expression of RIP3, p-MLKL and NLRP3 in pancreatic tissue of HP-MSCs group was significantly lower than those of SAP group and N-MSCs group (P＜0.05). Conclusions HP-MSCs can significantly alleviate SAP than N-MSCs, and the mechanism is potentially related to the inhibition of RIP1/RIP3-MLKL signaling pathway.

Key words: hypoxic preconditioning, placental mesenchymal stem cells, severe acute pancreatitis, receptor interacting protein 3(RIP3)

中图分类号:

R657.5+1

景光旭, 王张鹏, 刘忠钰, 吕双, 孙红玉. 低氧预处理胎盘间充质干细胞减轻重症急性胰腺炎模型小鼠胰腺组织损伤[J]. 基础医学与临床, 2023, 43(9): 1346-1352.

JING Guangxu, WANG Zhangpeng, LIU Zhongyu, LYU Shuang, SUN Hongyu. Hypoxic preconditioning placental mesenchymal stem cells alleviate pathological lesion in mouse models with severe acute pancreatitis[J]. Basic & Clinical Medicine, 2023, 43(9): 1346-1352.

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低氧预处理胎盘间充质干细胞减轻重症急性胰腺炎模型小鼠胰腺组织损伤

Hypoxic preconditioning placental mesenchymal stem cells alleviate pathological lesion in mouse models with severe acute pancreatitis

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