犬脂肪间充质干细胞及其外泌体修复庆大霉素致犬肾小管上皮细胞损伤

Canine adipose-derived mesenchymal stem cells and their exosomes in the repair of gentamicin-induced renal tubular epithelial cell injury

背景:犬肾脏损伤的特点是肾小管上皮细胞凋亡坏死。最近研究表明间充质干细胞及其外泌体在人、大鼠、小鼠肾损伤治疗中显示出良好的效果,但对犬的研究甚少。目的:探讨犬脂肪间充质干细胞及其外泌体对庆大霉素致犬肾小管上皮细胞损伤的影响。方法:采用5 mmol/L硫酸庆大霉素处理犬肾小管上皮细胞,随后分别将犬脂肪间充质干细胞及其条件培养基和外泌体与受损犬肾小管上皮细胞共培养。在24 h及48 h后采用CCK-8法测定各组细胞增殖活力,流式细胞术检测各组细胞凋亡率,最后通过Q-PCR法检测PCNA、Bcl-2和Bax基因的表达。结果与结论:犬脂肪间充质干细胞及其条件培养基和外泌体均可显著促进受损犬肾小管上皮细胞增殖以及减少细胞凋亡(P<0.05),其中犬脂肪间充质干细胞外泌体的效果最好,可显著提高受损犬肾小管上皮细胞PCNA、Bcl-2基因的表达(P<0.05),对庆大霉素诱导的犬肾小管上皮细胞损伤发挥修复作用。

BACKGROUND:Canine kidney injury is characterized by the apoptosis and necrosis of renal tubular epithelial cells.Recent developments in mesenchymal stem cells and their exosomes research have shown great promise for the treatment of kidney injury in humans,rats and mice,but little research has been done on dogs.OBJECTIVE:To investigate the effects of canine adipose-derived mesenchymal stem cells and their exosomes on canine renal tubular epithelial cell injury induced by gentamicin in vitro.METHODS:Canine renal tubular epithelial cells were treated by 5 mmol/L gentamicin sulfate.Subsequently,canine adipose-derived mesenchymal stem cells and their conditional medium and exosomes were co-cultured with damaged canine renal tubular epithelial cells respectively.After 24 and 48 hours,the cell proliferation activity of each group was measured by cell counting kit-8 method,and the apoptosis rate of each group was detected by flow cytometry.Finally,Q-PCR was used to further reveal the effects of canine adipose-derived mesenchymal stem cell exosomes on PCNA,Bcl-2 and Bax genes in these cells.RESULTS AND CONCLUSION:Canine adipose-derived mesenchymal stem cells,their conditioned media,and exosomes could significantly promote proliferation and reduce apoptosis in damaged canine renal tubular epithelial cells(P<0.05).Among them,canine adipose-derived mesenchymal stem cell exosomes worked best,which could significantly increase the expression of PCNA and Bcl-2 genes in damaged canine renal tubular epithelial cells(P<0.05).These results suggest that canine adipose-derived mesenchymal stem cells can repair the canine renal tubular epithelial cell damage induced by gentamicin through their exosomes.