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.

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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

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