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

Abstract

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

CLC Number:

S857.12

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.

References 30

[1]	何剑戈, 任爱红, 施懿诚, 等. 肝缺血再灌注损伤的机制及其预防[J]. 河南科技大学学报:医学版, 2012, 30(3):235-237.HE J G, REN A H, SHI Y C, et al. Mechanism and prevention of hepatic ischemia-reperfusion injury[J]. Journal of Henan University of Science & Technology:Medical Science, 2012, 30(3):235-237. (in Chinese)
[2]	REICH H, MCGLYNN F, DECAPRIO J, et al. Laparoscopic excision of benign liver lesions[J]. Obstet Gynecol, 1991, 78(5 Pt 2):956-958.
[3]	PANISELLO-ROSELLÓ A, ROSELLÓ-CATAFAU J. Molecular mechanisms and pathophysiology of ischemia-reperfusion injury[J]. Int J Mol Sci, 2018, 19(12):4093.
[4]	JIAO Z H, MA Y J, LIU X N, et al. Adipose-derived stem cell transplantation attenuates inflammation and promotes liver regeneration after ischemia-reperfusion and hemihepatectomy in swine[J]. Stem Cells Int, 2019, 2019:2489584.
[5]	李仕来, 尚丽明, 秦权林, 等. 橘皮苷通过激活Nrf2/HO-1通路增强自噬保护肝脏缺血再灌注损伤[J]. 中华中医药学刊, 2021, 39(4):105-108.LI S L, SHANG L M, QIN Q L, et al. Hesperidin induces autophagy to protect against liver ischemia/reperfusion injury via activation of the Nrf2/HO-1 pathway[J]. Chinese Archives of Traditional Chinese Medicine, 2021, 39(4):105-108. (in Chinese)
[6]	高伟东, 冯赞杰, 彭慈军, 等. 杜仲不同部位及不同提取方法的提取物对大鼠肝脏缺血再灌注损伤的保护作用[J]. 中国普通外科杂志, 2020, 29(8):958-965.GAO W D, FENG Z J, PENG C J, et al. Protective effects of extracts from different parts of Eucommia ulmoides and by different extraction methods against hepatic ischemia-reperfusion injury in rats[J]. Chinese Journal of General Surgery, 2020, 29(8):958-965. (in Chinese)
[7]	王双龙, 于美, 吕镔, 等. 声脉冲辐射力成像定量评估兔肝缺血再灌注损伤实验研究[J]. 陕西医学杂志, 2020, 49(2):140-143, 147.WANG S L, YU M, LV B, et al. Quantitative assessment of hepatic ischemia reperfusion injury with acoustic radiation force impulse in rabbit[J]. Shaanxi Medical Journal, 2020, 49(2):140-143, 147. (in Chinese)
[8]	董婧, 温吉昱, 陈玉婷, 等. 丙泊酚对老年犬心肌缺血再灌注损伤的影响[J]. 沈阳农业大学学报, 2018, 49(2):209-213.DONG J, WEN J Y, CHEN Y T, et al. The effects of propofol on myocardial ischemia reperfusion injury in aged canines[J]. Journal of Shenyang Agricultural University, 2018, 49(2):209-213. (in Chinese)
[9]	ABDO E E, CUNHA J E M, DELUCA P, et al. Protective effect of N2-mercaptopropionylglycine on rats and dogs liver during ischemia/reperfusion process[J]. Arq Gastroenterol, 2003, 40(3):177-180.
[10]	张千振, 葛延松, 焦智慧, 等. 小型猪腹腔镜肝缺血再灌注合并肝部分切除对肝细胞凋亡及相关基因影响的研究[J]. 中国兽医杂志, 2018, 54(4):22-25, 28.ZHANG Q Z, GE Y S, JIAO Z H, et al. Effect of laparoscopic ischemia reperfusion combined with partial hepatectomy on hepatocyte apoptosis and related gene expression in miniature pigs[J]. Chinese Journal of Veterinary Medicine, 2018, 54(4):22-25, 28. (in Chinese)
[11]	JIAO Z H, LIU X N, MA Y J, et al. Adipose-derived stem cells protect ischemia-reperfusion and partial hepatectomy by attenuating endoplasmic reticulum stress[J]. Front Cell Dev Biol, 2020, 8:177.
[12]	NAHAR S, NAKASHIMA Y, MIYAGI-SHIOHIRA C, et al. Cytokines in adipose-derived mesenchymal stem cells promote the healing of liver disease[J]. World J Stem Cells, 2018, 10(11):146-159.
[13]	JIAO Z H, MA Y J, ZHANG Q Z, et al. The adipose-derived mesenchymal stem cell secretome promotes hepatic regeneration in miniature pigs after liver ischaemia-reperfusion combined with partial resection[J]. Stem Cell Res Ther, 2021, 12(1):218.
[14]	LEE S K, LEE S C, KIM S J. A novel cell-free strategy for promoting mouse liver regeneration:utilization of a conditioned medium from adipose-derived stem cells[J]. Hepatol Int, 2015, 9(2):310-320.
[15]	VAN POLL D, PAREKKADAN B, CHO C H, et al. Mesenchymal stem cell-derived molecules directly modulate hepatocellular death and regeneration in vitro and in vivo[J]. Hepatology, 2008, 47(5):1634-1643.
[16]	CURTIS T M, HANNETT J M, HARMAN R M, et al. The secretome of adipose-derived mesenchymal stem cells protects SH-SY5Y cells from arsenic-induced toxicity, independent of a neuron-like differentiation mechanism[J]. Neurotoxicology, 2018, 67:54-64.
[17]	LI X T, MA T, SUN J C, et al. Harnessing the secretome of adipose-derived stem cells in the treatment of ischemic heart diseases[J]. Stem Cell Res Ther, 2019, 10(1):196.
[18]	NIADA S, GIANNASI C, GOMARASCA M, et al. Adipose-derived stromal cell secretome reduces TNFα-induced hypertrophy and catabolic markers in primary human articular chondrocytes[J]. Stem Cell Res, 2019, 38:101463.
[19]	AKHONDZADEH F, KADKHODAEE M, SEIFI B, et al. Adipose-derived mesenchymal stem cells and conditioned medium attenuate the memory retrieval impairment during sepsis in rats[J]. Mol Neurobiol, 2020, 57(9):3633-3645.
[20]	李辉, 葛延松, 白鸽, 等. 富氢生理盐水干预小型猪肝脏缺血再灌注合并肝脏部分切除损伤中内质网应激影响的研究[J]. 中国畜牧兽医, 2018, 45(6):1692-1699.LI H, GE Y S, BAI G, et al. Hydrogen-rich saline protects effect against liver ischemia-reperfusion injury and hepatectomy by inhibiting endoplasmic reticulum stress in miniature pig[J]. China Animal Husbandry & Veterinary Medicine, 2018, 45(6):1692-1699. (in Chinese)
[21]	UCHIDA Y, KE B B, FREITAS M C S, et al. The emerging role of T cell immunoglobulin mucin-1 in the mechanism of liver ischemia and reperfusion injury in the mouse[J]. Hepatology, 2010, 51(4):1363-1372.
[22]	LEE S C, KIM J O, KIM S J. Secretome from human adipose-derived stem cells protects mouse liver from hepatic ischemia-reperfusion injury[J]. Surgery, 2015, 157(5):934-943.
[23]	KIM Y H, CHO K A, PARK M, et al. Conditioned medium from tonsil-derived mesenchymal stem cells relieves CCl4-induced liver fibrosis in mice[J]. Tissue Eng Regen Med, 2019, 16(1):51-58.
[24]	葛延松. 小型猪ADSCs对腹腔镜肝脏I/R合并部分切除损伤保护作用的研究[D]. 哈尔滨:东北农业大学, 2019.GE Y S. Study of the protective effect of miniature pig adipose derived stem cells on hepatic ischemia-reperfusion combined with laparoscopic hepatectomy Injury[D]. Harbin:Northeast Agricultural University, 2019. (in Chinese)
[25]	张华. 腹腔镜技术建立小型猪肝损伤模型及肝脏环境变化的研究[D]. 哈尔滨:东北农业大学, 2015.ZHANG H. Study of the establishment of liver injury models by laparoscopic technique in miniture pigs and the changes of liver enviroment[D]. Harbin:Northeast Agricultural University, 2015. (in Chinese)
[26]	FRASER J R E, ALCORN D, LAURENT T C, et al. Uptake of circulating hyaluronic acid by the rat liver:cellular localization in situ[J]. Cell Tissue Res, 1985, 242(3):505-510.
[27]	GEORGE J, STERN R. Serum hyaluronan and hyaluronidase:very early markers of toxic liver injury[J]. Clin Chim Acta, 2004, 348(1-2):189-197.
[28]	CHEN Y X, ZENG Z C, SUN J, et al. Mesenchymal stem cell-conditioned medium prevents radiation-induced liver injury by inhibiting inflammation and protecting sinusoidal endothelial cells[J]. J Radiat Res, 2015, 56(4):700-708.
[29]	ABAZOV V M, ABBOTT B, ACHARYA B S, et al. Precision measurement of the ratio B(t→Wb)/B(t→Wq) and extraction of Vtb[J]. Phys Rev Lett, 2011, 107(12):121802.
[30]	ZANGI L, MARGALIT R, REICH-ZELIGER S, et al. Direct imaging of immune rejection and memory induction by allogeneic mesenchymal stromal cells[J]. Stem Cells, 2009, 27(11):2865-2874.

Study of Adipose-derived Stem Cell Condition Medium on the Inflammatory Response of Miniature Pigs with Laparoscopic Liver Ischemia Reperfusion Combined with Partial Hepatectomy

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