Background
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)].
Methods
Animals (n = 24) were categorized into group 1 (sham-operated control), group 2 (DKD), group 3 [DKD + HUCDMSCs (2.1 × 105/IRA injection at day 28 after CKD induction)] and group 4 [(DKD + HUCDMSCs (6.3 × 105/IRA injection)].
Results
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.
Conclusion
IRA injection of xenogeneic MSCs was safe and effectively protected the residual renal function and architectural integrity in DKD rat.
This study tested the impact of single dose and two doses of endothelial progenitor cells (EPCs) and EPCs-derived condition medium (CM) on protecting the left-ventricular myocardium (LVM) from acute ischemia-reperfusion (IR) injury. In vitro study showed EPCs and CM had comparably higher capacity for enhancement of angiogenesis as compared with the controls (all P < .001). Adult-male SD rats ( n = 36) were equally categorized into groups 1 (sham-operated control), 2 (IR+vehicle), 3 [IR+EPCs/1.2 × 106/intravenous administration at 3 h after IR procedure), 4 (IR+EPCs/1.2 × 106/at 3 h/24 h after IR), 5 (IR+CM/3.0cc/intravenous administration at 3 h after IR), 6 (IR+EPCs/3.0cc/at 3h/24 h after IR), and euthanized by day 3 after IR. The left-ventricular-ejection-fraction, protein and cellular expressions of endothelial-cell markers (CD31/vWF), small vessel number and protein expression of mitochondrial (mitochondrial-cytochrome-C) integrity were highest in group 1, lowest in group 2, significantly higher in group 4 than in groups 3/5/6 and significantly higher in groups 3/6 than in group 5 but they showed no differences in groups3/6, whereas the protein expressions of apoptotic (cleaved-caspase 3/cleaved-PARP), fibrotic (Smad3/TGF-ß), mitochondrial-damaged (cytosolic-cytochrome-C), heart-failed/pressure-overload (BNP), oxidative-stress (p47phox/NOX-1/NOX-2/oxidized protein), and autophagic (LCB3-II/LCB3-I) biomarkers and fibrotic/collagen-deposition areas exhibited an opposite pattern to endothelial-cell markers (all P < .0001). The protein expressions of angiogenesis (VEGF/SDF-1α/CXCR4/HIF-1α) were lowest in group 1, highest in group 4, significantly higher in groups 3/6 than in groups 2/5, significantly higher in group 5 than in group 2, but they showed no difference between groups 3/6 (all P < .0001). These results demonstrate that two consecutive doses of EPC/CM were superior to just one at protecting LVM against IR injury.
In this study we have examined whether hyperbaric oxygen (HBO)-assisted melatonin (Mel) therapy effectively preserves the heart function against ischemia (40-min)-reperfusion (IR) injury. First, the in vitro study has been performed by use of cell culture. H9C2 cells were treated as groups: A (H9C2) (without any treatment), B (H9C2+IR), C (H9C2+IR+HBO), D [H9C2+IR+Melatonin (50 µM)] and E (H9C2+IR+Melatonin+HBO). The result showed that the protein expressions of oxidative-stress (NOX-1/NOX-2)/inflammatory (TNF-α/NF-κB)/apoptotic (mitochondrial-Bax/cleaved-caspase-3/cleaved-PARP) and cellular levels of DNA/mitochondrial-damaged (γ-H2AX/XRCC1-CD90+/cytosolic-cytochrome-C) biomarker were significantly increased in group B compared to control group A. These biomarkers were significantly reduced in group C, D and E compared to groups B with the significantly higher reduction in group E than that in groups C and D (p<0.001). In the in vivo study, adult-male SD rats (n=40) were equally divided into group 1 (sham-operated-control), group 2 (IR), group 3 (IR+HBO therapy at 1.5/24/48h after IR procedure), group 4 [IR+Mel (50mg/kg at 1.5h, followed by 20mg/kg and at days 1/2/3 after IR)] and group 5 (IR+HBO-Mel) and the heart was harvested at 72h after IR. The result showed that at 72h, the circulating levels of endothelial-progenitor-cells (c-kit-CD31+/CD31-sca-1+/KDR-CD34+/VE-Cadherin-CD34+) were lowest in group 2, highest in group 5 and followed by groups 3 > 4 >1. The significant differences were present between each two matched groups (p<0.0001). The protein expressions of angiogenic factors (SDF-1α/CXCR4/VEGF/HIF-α) were progressively increased from groups 1 to 5 with significant differences. The protein expressions of apoptosis (mitochondrial-Bax/cleaved-caspase-3/cleaved-PARP)/fibrosis (TGF-ß/Smad3)/oxidative-stress (NOX-1/NOX-2/oxidized protein)/inflammation (TNF-α/IL-1ß/MMP-9) and infarct/fibrotic areas were significantly increased in group 2 compared to the control group 1. All these parameters were significantly reduced in groups 3-5 compared to group 2 with significantly lowest level in group 5 among groups 3-5 (p < 0.01), whereas the left-ventricular-ejection-fraction exhibited an opposite pattern compared to the inflammatory factors. In conclusion, HBO-Mel therapy offered a synergic benefit for protecting the heart from IR injury.
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