脂肪间充质干细胞对小型猪肝缺血再灌注合并肝切除组织细胞焦亡的影响

doi:10.11843/j.issn.0366-6964.2024.01.033

畜牧兽医学报 ›› 2024, Vol. 55 ›› Issue (1): 355-364.doi: 10.11843/j.issn.0366-6964.2024.01.033

脂肪间充质干细胞对小型猪肝缺血再灌注合并肝切除组织细胞焦亡的影响

马亚军^1,2, 焦智慧^1,2, 刘笑凝^1,2, 陆翔羽^1,2, 刘涛^1,2, 王月^1,2, 朴晨曦^1,2, 王洪斌^1,2*

1. 东北农业大学动物医学学院, 哈尔滨 150030;
2. 黑龙江省普通高等学校动物普通疾病防治重点实验室, 哈尔滨 150030

收稿日期:2023-06-12 出版日期:2024-01-23 发布日期:2024-01-24
通讯作者: 王洪斌,主要从事腹腔镜微创手术研究,E-mail:hbwang1940@neau.edu.cn
作者简介:马亚军(1994-),女,河北大城人,博士生,主要从事腹腔镜微创手术研究,E-mail:mayajun1994@126.com
基金资助:
国家自然科学基金项目（31972757）

Effects of Adipose-derived Mesenchymal Stem Cells on Pyroptosis of Miniature Pigs with Hepatic Ischemia-Reperfusion Combined with Hepatectomy

MA Yajun^1,2, JIAO Zhihui^1,2, LIU Xiaoning^1,2, LU Xiangyu^1,2, LIU Tao^1,2, WANG Yue^1,2, PIAO Chenxi^1,2, WANG Hongbin^1,2*

1. College of Animal Medicine, Northeast Agricultural University, Harbin 150030, China;
2. Heilongjiang Key Laboratory for Animals and Comparative Medicine, Harbin 150030, China

Received:2023-06-12 Online:2024-01-23 Published:2024-01-24

摘要/Abstract

摘要： 本研究旨在探究脂肪间充质干细胞（adipose derived mesenchymal stem cells， ADSCs）对小型猪肝缺血再灌注损伤（ischemia-reperfusion injury， IRI）合并肝切除组织细胞焦亡的影响。将18头巴马小型猪随机平分成3组，每组6头，分别为假手术组（sham组）、模型组（IRI组）、异体移植ADSCs干预组（ADSCs组，剂量为1×10⁶ cells·kg^-1），通过腹腔镜微创技术建立肝IRI合并肝切除损伤模型，于术后即刻分别向IRI组和ADSCs组小型猪肝实质注射生理盐水和ADSCs。于术后1、3和7 d采集血液和肝组织样本。应用ELISA法对血清中促炎因子白细胞介素18（interleukin-18， IL-18）、白细胞介素1β（interleukin-1β， IL-1β）的含量进行测定，应用RT-qPCR技术和Western blot技术对细胞焦亡相关基因与蛋白进行检测。结果显示，术后1、3 d时，相比于sham组，IRI组血清中促炎因子IL-18、IL-1β含量显著增加（P<0.01），肝组织中IL-18、IL-1β基因与蛋白表达量同样显著升高（P<0.01）；肝组织中细胞焦亡相关因子NOD样受体热蛋白结构域相关蛋白3（NOD-like receptor protein 3， NLRP3）、凋亡相关斑点样蛋白（apoptosis-associated speck-like protein containing a CARD， ASC）、Caspase-1、GSDMD和核因子κBp65（nuclear factor kappa-Bp65， NF-κBp65）基因与蛋白水平均显著升高（P<0.05）。经ADSCs干预后，血清和肝组织中IL-18、IL-1β表达量显著降低（P<0.05），肝组织中细胞焦亡相关因子NLRP3、ASC、GSDMD和NF-κBp65基因与蛋白水平均显著降低（P<0.05）。综上，本研究证明，小型猪腹腔镜肝IRI合并肝切除损伤会诱导细胞焦亡的发生，而ADSCs能够降低及改善这种肝损伤引起的组织细胞焦亡损伤。

关键词: 脂肪间充质干细胞, 肝缺血再灌注损伤, 细胞焦亡, 腹腔镜, 小型猪

Abstract: The purpose of this study was to investigate the effect of adipose derived mesenchymal stem cells (ADSCs) on pyroptosis of miniature pigs with liver Ischemia-reperfusion injury (IRI) combined with hepatectomy. Eighteen Bama mini pigs were randomly divided into three groups, 6 in each group, which were Sham surgery group (sham group), model group (IRI group) and ADSCs intervention group (ADSCs group, dose is 1×10⁶ cells·kg^-1). The model of liver IRI combined with hepatectomy injury was established by laparoscopic minimally invasive technique. The IRI group and ADSCs group mini pigs were injected with normal saline and ADSCs into the liver parenchyma immediately after operation. Blood and liver tissue samples were collected 1, 3, and 7 d after surgery. Application of ELISA method to detect pro-inflammatory factors IL-18 and IL-1β in serum. The content of pyroptosis related genes and proteins were detected by RT-qPCR and Western blot. The results showed that on the 1 and 3 d, compared to the sham group, the serum pro-inflammatory factors IL-18 and IL-1β in the IRI group were significantly higher than those in the sham group (P<0.01). The expression levels of IL-18 and IL-1β genes and proteins in liver tissue were also significantly increased (P<0.01); The levels of the pyroptosis related factors NOD-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), Caspase-1, GSDMD and nuclear factor kappa-Bp65 (NF-κBp65) genes and proteins in liver tissue were significantly elevated (P<0.05). After intervention with ADSCs, the expression levels of IL-18 and IL-1β in serum and liver tissue were significantly reduced (P<0.05). The gene and protein expression of pyroptosis related factors NLRP3, ASC, Caspase-1, GSDMD and NF-κBp65 in liver tissue was significantly reduced (P<0.05). Therefore, this study proves that laparoscopic liver IRI combined with hepatectomy injury can induce pyroptosis in miniature pigs, and ADSCs can reduce and improve the pyroptosis injury caused by this liver injury.

Key words: adipose derived mesenchymal stem cells, liver ischemia-reperfusion injury, pyroptosis, laparoscope, miniature pig

中图分类号:

S854

马亚军, 焦智慧, 刘笑凝, 陆翔羽, 刘涛, 王月, 朴晨曦, 王洪斌. 脂肪间充质干细胞对小型猪肝缺血再灌注合并肝切除组织细胞焦亡的影响[J]. 畜牧兽医学报, 2024, 55(1): 355-364.

MA Yajun, JIAO Zhihui, LIU Xiaoning, LU Xiangyu, LIU Tao, WANG Yue, PIAO Chenxi, WANG Hongbin. Effects of Adipose-derived Mesenchymal Stem Cells on Pyroptosis of Miniature Pigs with Hepatic Ischemia-Reperfusion Combined with Hepatectomy[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 355-364.

参考文献 13

[1]	LI J, LI R J, LV G Y, et al.The mechanisms and strategies to protect from hepatic ischemia-reperfusion injury[J].Eur Rev Med Pharmacol Sci, 2015, 19(11):2036-2047.
[2]	BERGSBAKEN T, FINK S L, COOKSON B T.Pyroptosis:host cell death and inflammation[J].Nat Rev Microbiol, 2009, 7(2):99-109.
[3]	DING J J, WANG K, LIU W, et al.Pore-forming activity and structural autoinhibition of the gasdermin family[J].Nature, 2016, 535(7610):111-116.
[4]	XIA S Y, HOLLINGSWORTH IV L R, WU H.Mechanism and regulation of gasdermin-mediated cell death[J].Cold Spring Harbor Perspect Biol, 2020, 12(3):a036400.
[5]	BERGSBAKEN T, FINK S L, COOKSON B T.Pyroptosis:host cell death and inflammation[J].Nat Rev Microbiol, 2009, 7(2):99-109.
[6]	DU Y C, ZHONG F R, CHENG H L, et al.The dietary supplement γ-oryzanol attenuates hepatic ischemia reperfusion injury via inhibiting endoplasmic reticulum stress and HMGB1/NLRP3 inflammasome[J].Oxid Med Cell Longev, 2021, 2021:4628050.
[7]	TSUNG A, SAHAI R, TANAKA H, et al.The nuclear factor HMGB1 mediates hepatic injury after murine liver ischemia-reperfusion[J].J Exp Med, 2005, 201(7):1135-1143.
[8]	COLL R C, SCHRODER K, PELEGRÍN P.NLRP3 and pyroptosis blockers for treating inflammatory diseases[J].Trends Pharmacol Sci, 2022, 43(8):653-668.
[9]	SHI C Y, WANG Q, RAO Z Q, et al.Diabetes induces hepatocyte pyroptosis by promoting oxidative stress-mediated NLRP3 inflammasome activation during liver ischaemia and reperfusion injury[J].Ann Transl Med, 2020, 8(12):739.
[10]	BIANCO P."Mesenchymal" stem cells[J].Ann Rev Cell Dev Biol, 2014, 30:677-704.
[11]	PRIETO GONZÁLEZ E A.Heterogeneity in adipose stem cells[M]//BIRBRAIR A.Stem Cells Heterogeneity-Novel Concepts.Cham:Springer, 2019:119-150.
[12]	KONNO M, HAMABE A, HASEGAWA S, et al.Adipose-derived mesenchymal stem cells and regenerative medicine[J].Dev Growth Differ, 2013, 55(3):309-318.
[13]	AL-GHADBAN S, BUNNELL B A.Adipose tissue-derived stem cells:immunomodulatory effects and therapeutic potential[J].Physiology (Bethesda), 2020, 35(2):125-133.

[1]	焦广明, 吕英光, 桑金芳, 寇志鹏, 刘涛, 王月, 陆翔宇, 朴晨曦, 马亚军, 张建涛, 王洪斌. 脂肪间充质干细胞与甲泼尼龙联合用药对小型猪异体皮肤移植的影响[J]. 畜牧兽医学报, 2023, 54(8): 3533-3545.
[2]	连玉举, 张致远, 廖晓波, 魏红江, 印遇龙, 刘梅. 医用小型猪选育方法和应用进展[J]. 畜牧兽医学报, 2023, 54(7): 2667-2682.
[3]	孙浩, 者小书, 张文奇, 郝斐, 李文楠, 刘洁, 刘东军. 毛壳素对绒山羊ADSCs组蛋白甲基化修饰的影响研究[J]. 畜牧兽医学报, 2022, 53(7): 2380-2389.
[4]	田威龙, 司景磊, 刘笑笑, 綦文晶, 陈奎蓉, 程锋, 李月月, 吕冬玲, 梁靓, 高九昱, 奉玲丽, 莫家远, 兰干球, 梁晶. 高脂高糖饮食对小型猪肠道微生物的影响[J]. 畜牧兽医学报, 2022, 53(4): 1143-1153.
[5]	梁文清, 刘忠华, 常晓月, 何婷, 陈嘉, 马晓莉, 唐兆新, 余文兰. 长期高铜暴露通过影响线粒体自噬和细胞焦亡诱导大鼠肝组织损伤[J]. 畜牧兽医学报, 2022, 53(12): 4490-4500.
[6]	石玉华, 傅心雨, 郑梦洁, 吕倩, 师福山. 非经典细胞焦亡体外模型的构建[J]. 畜牧兽医学报, 2021, 52(8): 2284-2290.
[7]	焦智慧, 张千振, 王月, 刘涛, 刘博洋, 马亚军, 刘笑凝, 朴晨曦, 王洪斌. 脂肪来源间充质干细胞条件培养基对小型猪腹腔镜肝损伤氧化应激反应的影响[J]. 畜牧兽医学报, 2021, 52(6): 1734-1743.
[8]	焦智慧, 马亚军, 刘笑凝, 张千振, 王月, 刘涛, 朴晨曦, 刘博洋, 王洪斌. 脂肪间充质干细胞条件培养基对小型猪肝缺血再灌注合并肝部分切除炎症反应的影响[J]. 畜牧兽医学报, 2021, 52(12): 3651-3659.
[9]	聂雪伊, 刘蕾, 郑雪迪, 杨易, 徐金瑞, 王玉炯. BCG感染巨噬细胞后内质网应激对细胞焦亡的调控作用[J]. 畜牧兽医学报, 2021, 52(10): 2842-2851.
[10]	高泽乾，朱学亮，张志东，窦永喜. 病毒感染激活炎症小体的分子机制[J]. 畜牧兽医学报, 2016, 47(11): 2167-2174.
[11]	李完波，朱亚玲，艾华水，郭添福. 猪小体型相关受选择基因位点的研究[J]. 畜牧兽医学报, 2016, 47(10): 1977-1985.
[12]	李新，姜胜，侯金龙，范宏刚，王洪斌. 小型猪复合麻醉剂对大鼠中枢神经系统p-p38MAPK蛋白表达的影响[J]. 畜牧兽医学报, 2015, 46(7): 1253-1258.
[13]	李妍，侯金龙，牛栋梁，姜胜，时静，范宏刚，王洪斌. 小型猪复合麻醉颉颃剂对大鼠各脑区p-p38蛋白及c-myc mRNA表达的影响[J]. 畜牧兽医学报, 2015, 46(2): 317-322.
[14]	刘正亚，徐学文，方勇，张丹丹，任莹，崔敏，赵胜军. 两种安装多位点血管瘘手术方法对山羊血清蛋白质含量、肝功能及营养物质消化和代谢的影响[J]. 畜牧兽医学报, 2014, 45(7): 1129-1134.
[15]	牛栋梁，姜胜，李新，陈皓，范宏刚，王洪斌. 小型猪专用复合麻醉剂对大鼠不同脑区β-arrestin2基因mRNA转录的影响[J]. 畜牧兽医学报, 2014, 45(5): 839-843.

脂肪间充质干细胞对小型猪肝缺血再灌注合并肝切除组织细胞焦亡的影响

Effects of Adipose-derived Mesenchymal Stem Cells on Pyroptosis of Miniature Pigs with Hepatic Ischemia-Reperfusion Combined with Hepatectomy

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献 13

相关文章 15

编辑推荐

Metrics

本文评价