Institute of Nephrology and Blood Purification, The First Affiliated Hospital of Jinan University, Jinan University, Guangzhou, 510632, China.
Department of Cardiology, The First Affiliated Hospital, Jinan University, Guangzhou, 510632, China.
Stem Cell Res Ther. 2022 May 7;13(1):186. doi: 10.1186/s13287-022-02857-5.
This experimental study was designed as a preclinical study for testing the hypothesis that intrarenal arterial (IRA) transfusion of human umbilical cord-derived mesenchymal stem cells (HUCDMSCs) therapy preserved the residual renal function of diabetic kidney disease (DKD) in rat [induction by 5/6 nephrectomy of left kidney and right nephrectomy, followed by intraperitoneal administration of aminoguanidine (180 mg/kg) and streptozotocin (30 mg/kg)].
Animals (n = 24) were categorized into group 1 (sham-operated control), group 2 (DKD), group 3 [DKD + HUCDMSCs (2.1 × 10/IRA injection at day 28 after CKD induction)] and group 4 [(DKD + HUCDMSCs (6.3 × 10/IRA injection)].
By day 60 after DKD induction, the kidneys were harvested and the result showed that the creatinine level, ratio of urine protein/urine creatinine and kidney injury score were lowest in group 1, highest in group 2 and significantly lower in group 4 than in group 3 (all p < 0.0001). The protein expressions of apoptotic (cleaved caspase-3/cleaved PARP/mitochondrial Bax), fibrotic (TGF-ß/p-Smad3), autophagic (ratio of LC3B-II/LC3B-I, Atg5/Beclin-1), oxidative stress (NOX-1/NOX-2/oxidized protein/p22phox), mitochondrial/DNA-damaged (cytosolic-cytochrome-C/DRP1/γ-H2AX) and inflammatory (MMP-9/TNF-α/p-NF-κB) biomarkers exhibited an identical pattern, whereas the protein expressions of angiogenesis factors (CD31/vWF/vascularity) exhibited an opposite pattern of creatinine level among the groups (all p < 0.0001). Histopathological findings demonstrated the renal tubular-damaged (KIM-1)/kidney fibrosis area/oxidative stress (8-OHdG + cells) expressed an identical pattern, whereas the podocyte components (ZO-1/synaptopodin/podocin) exhibited an opposite pattern of creatinine level among the groups (all p < 0.0001). No tumorigenesis or immune rejection event was identified.
IRA injection of xenogeneic MSCs was safe and effectively protected the residual renal function and architectural integrity in DKD rat.
本实验研究旨在进行临床前研究，以验证以下假设：经肾内动脉（IRA）输注人脐带间充质干细胞（HUCDMSCs）治疗可保留糖尿病肾病（DKD）大鼠的残余肾功能[通过左肾 5/6 切除术和右肾切除术诱导，随后腹腔内给予氨基胍（180mg/kg）和链脲佐菌素（30mg/kg）]。
动物（n=24）分为 4 组：第 1 组（假手术对照）、第 2 组（DKD）、第 3 组[DKD+HUCDMSCs（2.1×10 个/IRA 注射，在 CKD 诱导后 28 天）]和第 4 组[DKD+HUCDMSCs（6.3×10 个/IRA 注射]。
DKD 诱导后 60 天，收获肾脏，结果显示，第 1 组肌酐水平、尿蛋白/尿肌酐比值和肾脏损伤评分最低，第 2 组最高，第 4 组明显低于第 3 组（均 p<0.0001）。凋亡（cleaved caspase-3/cleaved PARP/mitochondrial Bax）、纤维化（TGF-β/p-Smad3）、自噬（LC3B-II/LC3B-I 比值、Atg5/Beclin-1）、氧化应激（NOX-1/NOX-2/氧化蛋白/p22phox）、线粒体/DNA 损伤（细胞质细胞色素-C/DRP1/γ-H2AX）和炎症（MMP-9/TNF-α/p-NF-κB）生物标志物的蛋白表达呈现相同的模式，而各组间血管生成因子（CD31/vWF/vascularity）的蛋白表达与肌酐水平呈相反的模式（均 p<0.0001）。组织病理学检查显示肾小管损伤（KIM-1）/肾纤维化面积/氧化应激（8-OHdG+细胞）的表达呈现相同的模式，而足细胞成分（ZO-1/synaptopodin/podocin）的表达与肌酐水平呈相反的模式（均 p<0.0001）。未发现肿瘤发生或免疫排斥事件。
肾内动脉注射异种间充质干细胞是安全的，可有效保护 DKD 大鼠的残余肾功能和结构完整性。
