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

doi:10.11843/j.issn.0366-6964.2021.012.031

摘要/Abstract

摘要： 本研究旨在探究脂肪间充质干细胞条件培养基（adipose-derived mesenchymal stem cells-condition medium，ADSCs-CM）对小型猪肝缺血再灌注合并肝部分切除炎症反应的作用。基于腹腔镜技术对24头小型猪建立肝缺血再灌注（ischemia reperfusion，IR）合并肝部分切除模型，根据术后对肝移植4种不同物质：生理盐水、浓缩的基础培养基、浓缩的脂肪间充质干细胞培养基、脂肪间充质干细胞，将小型猪分为模型组（IRI）、DMEM组（DMEM）、ADSCs-CM组（CM）和ADSCs组（ADSCs）4组，每组6只。分别于术前、术后1、3、7 d采集血液与肝组织样本，通过病理组织学炎性细胞的观察，血液常规指标的检测，血清皮质醇（COR）、C反应蛋白（CRP）、透明质酸（HA）的ELISA检测，肝组织炎症相关基因IL-1β、IL-6、TNF-α、IL-10 mRNA的qRT-PCR检测综合评价ADSCs-CM对炎症反应的作用。结果显示：术后1和3 d：IRI组、DMEM组的病理切片出现较多炎性细胞，血常规和炎症相关基因结果也表明术后发生炎症反应，而CM组和ADSCs组显著减少组织中炎性细胞的浸润和血液中白细胞（WBC）、中性粒细胞（NE）、淋巴细胞（LY）的细胞数，降低组织中促炎因子IL-1β、IL-6、TNF-α mRNA的表达水平，并提高抗炎因子IL-10 mRNA的表达水平。术后7 d，各组基本恢复到术前水平。综上表明：肝缺血再灌注合并肝部分切除可致炎症的发生，ADSCs-CM和ADSCs均可改善肝缺血再灌注合并肝部分切除的炎症反应，ADSCs-CM移植有潜力成为未来治疗炎症反应的无细胞疗法。

关键词: 小型猪, 腹腔镜, 缺血再灌注, 肝部分切除, ADSCs-CM, 炎症

Abstract: The aim of this study was to explore the efficacy of adipose-derived mesenchymal stem cells condition medium (ADSCs-CM) on the inflammatory response of liver ischemia reperfusion combined with partial hepatectomy in miniature pigs. Based on the laparoscopic technique, a model of liver ischemia reperfusion (IR) combined with partial hepatectomy was established in 24 miniature pigs. Miniature pigs were divided into four groups:model group (IRI), DMEM group (DMEM), ADSCs-CM group (CM) and ADSCs group (ADSCs), with six pigs in each group, according to the four different substances namely physiological saline, concentrated basal medium, concentrated ADSCs-CM, ADSCs transplanted to the liver after surgery. The blood and liver tissue samples were harvested pre-operation, 1 day, 3 days, and 7 days after operation. The effect of ADSCs-CM on inflammation were comprehensively evaluated through histopathological inflammatory cell observation, blood indicators detection, serum cortisol (COR), C-reactive protein (CRP) and hyaluronic acid (HA) detection of ELISA, liver tissue inflammation-related IL-1β, IL-6, TNF-α, IL-10 mRNA detection of qRT-PCR. The results found that 1 and 3 days after the operation:there were more inflammatory cells in the pathological tissue sections of the IRI group and DMEM group. The results of blood routine and inflammation-related genes also showed that there was obvious inflammation after the operation. The ADSCs-CM and ADSCs treatment groups significantly reduced the infiltration of inflammatory cells in the tissues and the number of white blood cells (WBC), neutrophils (NE) and lymphocytes (LY) in the blood, and decreased the expression levels of pro-inflammatory factor IL-1β, IL-6, TNF-α mRNA in tissues, and increased the expression level of anti-inflammatory factor IL-10 mRNA. After 7 days, each group basically recovered to the pre-operation. The results showed that liver ischemia-reperfusion combined with partial hepatectomy can cause inflammation, and both ADSCs-CM and ADSCs improved the inflammatory response of liver ischemia-reperfusion combined with partial hepatectomy. Transplantation of ADSCs-CM has the potential to become a cell-free therapy for the treatment of inflammation in the future.

Key words: miniature pigs, laparoscopy, ischemia-reperfusion, partial hepatectomy, ADSCs-CM, inflammation

中图分类号:

S857.12

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.

JIAO Zhihui, MA Yajun, LIU Xiaoning, ZHANG Qianzhen, WANG Yue, LIU Tao, PIAO Chenxi, LIU Boyang, WANG Hongbin. Study of Adipose-derived Stem Cell Condition Medium on the Inflammatory Response of Miniature Pigs with Laparoscopic Liver Ischemia Reperfusion Combined with Partial Hepatectomy[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(12): 3651-3659.

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