摘要
为探究脂肪间充质干细胞分泌蛋白(adipose-derived mesenchymal stem cells-secretome,ADSCs-secretome)对小型猪腹腔镜肝缺血再灌注损伤(ischemia reperfusion injury,IRI)合并肝部分切除术肝细胞自噬损伤修复的影响,本试验选取24头健康小型猪,随机分为4组(n=6),分别为IRI组、DMEM组、ADSCs组和ADSCs-sec组。各组分别于术前、术后3 d、术后7 d通过腹腔镜技术采取肝脏组织样本。结果显示,术后3 d,DMEM组与IRI组相比,自噬相关基因(Beclin-1、ATG5、ATG12、p62)与蛋白(Beclin-1、LC3Ⅱ、p62),自噬通路相关基因(PI3K、Akt、mTOR)与蛋白比值(p-Akt/Akt、p-mTOR/mTOR)表达水平无显著性差异(P>0.05);ADSCs-sec组与DMEM组相比,Beclin-1、ATG5、ATG12 mRNA与Beclin-1、LC3Ⅱ蛋白表达水平显著性降低(P<0.01),p62 mRNA与蛋白表达水平显著性升高(P<0.01),PI3K、Akt、mTOR mRNA与p-Akt/Akt、p-mTOR/mTOR比值表达水平显著性升高(P<0.01)。术后7 d,各组之间数据无显著性差异(P>0.05)。免疫组化结果显示,检测LC3Ⅱ蛋白结果与Western blot结果相一致。结果表明,ADSCs-secretome能够促进肝IRI合并肝部分切除肝细胞过度自噬损伤修复,且可能是通过激活自噬负调控通路PI3K/Akt/mTOR进行调节的。
This study aims to investigate the effect of adipose-derived mesenchymal stem cells secretome(ADSCs-secretome)on the repair of autophagic injury during laparoscopic hepatic ischemia reperfusion injury(IRI)combined with partial hepatectomy in miniature pigs.Twenty-four healthy minipigs were selected and randomly divided into four groups(n=6):IRI group,DMEM group,ADSCs group and ADSCs-sec group.For each group,liver tissue samples were taken by laparoscopic surgery 3 d before surgery,and 3,7 d after surgery.The results showed that at 3 d after surgery,there was no significant difference in the expression levels of autophagy-related genes(Beclin-1,ATG5,ATG12,p62)and autophagy-related proteins(Beclin-1,LC3Ⅱ,p62),autophagy pathway-related genes(PI3 K,Akt,mTOR)and proteins(p-Akt/Akt,p-mTOR/mTOR)between the DMEM group and the IRI group(P>0.05);the expression levels of Beclin-1,ATG5,ATG12 mRNA and Beclin-1,LC3Ⅱproteins in the ADSCs-sec group were significantly decreased(P<0.01)and the expression levels of p62,PI3 K,Akt and mTOR mRNA and p62,p-Akt/Akt and pmTOR/mTOR proteins were significantly increased(P<0.01)compared with the DMEM group.At 7 dafter surgery,there was no significant difference in data between groups(P>0.05).Immunohistochemical results showed that the detection results of LC3Ⅱprotein were consistent with the Western blot results.The results showed that ADSCs-secretome could promote the repair of injury caused by excessive autophagy in hepatic IRI combined with partial hepatectomy,which may be regulated by activating the negative regulation pathway of autophagy PI3 K/Akt/mTOR.
作者
马亚军
刘笑凝
焦智慧
朴晨曦
徐嘉元
张千振
王洪斌
MA Yajun;LIU Xiaoning;JIAO Zhihui;PIAO Chenxi;XU Jiayuan;ZHANG Qianzhen;WANG Hong-bin(Heilongjiang Key Laboratory for Laboratory Animals and Comparative Medicine istry of Education,ColLege of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2021年第9期1828-1834,1856,共8页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(31972757,31772807)。
