Homing of PKH-26 labeled hATMSCs to the kidney tissue.

<p>(<b>A</b>) The representative of PKH-26 label (<b>B</b>) The staining for nuclei using (DAPI) (<b>C</b>) The overlay of (A) and (B) showed homing of PKH-26 labeled hATMSCs in renal interstitial tissue. PKH-26 staining merged with nuclear staining (in the circle), indicating PKH-26 labeled hATMSCs. (Original magnification x400 in A through C) (<b>D</b>) The quantization of PKH-26 labeled hATMSCs. Labeled cell was more frequently detected in CsA treated group compared to VH group. hATMSCs; human adipose tissue derived mesenchymal stem cells DAPI; 4′,6-Diamidino-2-Phenylindole.</p

Experimental design in this study.

<p>CsA was administered at a dose of 7.5 mg/kg subcutaneously daily. hATMSCs was infused at 0,1,2 and 3 weeks from the start of CsA with cell number of 3×10⁶/mL via tail vein. hATMSCs; human adipose tissue derived mesenchymal stem cells.</p

Effect of hATMSCs infusion on inflammatory cell infiltration in rats with chronic CsA nephropathy.

<p>(<b>A</b>) Representative photomicrographs of the immunohistochemical detection of ED1 protein (Original magnification x200). (<b>B</b>) Quantitative analysis of ED1-positive cells in the four groups. CsA administration increased ED1-positive cell number compared to VH and treatment with hATMSCs did not decrease it. hATMSCs; human adipose tissue derived mesenchymal stem cells.</p

The effects of treatment with hATMSC on basic parameters in experimental groups.

<p>ΔBody weight, changes in body weight; Scr, serum creatinine; ClCr, creatinine clearance; CsA, cyclosporine, hATMSC; human adipose tissue derived mesenchymal stem cell.</p>*<p>P<0.05 vs. VH group.</p>**<p>P<0.05 vs. CsA group.</p

The results of in vitro study.

<p>(<b>A</b>) The number of Annexin V positive cell didn’t increase in HK-2 cell cultured in hATMSCs- conditioned medium compared to HK-2 cell cultured in DMEM. Addition of CsA increased Annexin V positive cell both in DMED and hATMSCs-conditioned medium, and it was more frequently detected in HK-2 cell cultured in hATMSCs-conditioned medium compared to HK-2 cells cultured in DMEM. (<b>B</b>) The production of NO⁻ was increased in medium from hATMSCs treated with CsA compared with medium from hATMSCs cultured without CsA. (<b>C</b>) The 8-OHdG level was significantly increased in the medium from hATMSCs treated with CsA compared to medium from hATMSCs cultured withCsA-free media as well. hATMSCs; human adipose tissue derived mesenchymal stem cells.</p

Flowcytometric analysis of surface markers on cultured hATMSCs.

<p>(<b>A</b>) MSC-specific markers CD29, CD73, CD90 and CD105 were expressed on culture hATMSCs. (<b>B</b>) Hematopoietic stem cell markers CD31, CD34 and CD45 were not expressed.</p

Effect of hATMSCs infusion on oxidative damage in rats with chronic CsA nephropathy.

<p>(<b>A</b>) The serum and (<b>B</b>) urinary concentrations of 8-OHdG was increased in the CsA group, and further increased with the treatment of hATMSCs. hATMSCs; human adipose tissue derived mesenchymal stem cells.</p

Effect of MK-0626 on apoptosis during tacrolimus-induced pancreatic islet injury.

<p>In situ TdT-mediated dUTP–biotin nick end labeling (TUNEL) assay (A) and its analysis (B) to detect apoptosis in pancreatic islets in the experimental groups. MK-0626 (M) treatment significantly reduces TUNEL-positive cells (arrows) compared with tacrolimus (TAC) group. (C) Quantitative analysis in islets of the active form of caspase-3 in the experimental groups. Note that the MK-0626 treatment significantly reduced active caspase-3 compared with the TAC group. Original magnifications, x400. <i>n</i> = 8 rats per group. ^#P<0.05 vs. control. ^$P<0.05 vs. TAC.</p

Effect of MK-0626 on the expression of 8-hydroxy-2′-deoxyguanosine, manganese superoxide dismutase and heme oxygenase-1 during tacrolimus-induced pancreatic islet injury.

<p>(A and B) Immunohistochemistry for 8-hydroxy-2′-deoxyguanosine (8-OHdG) and its quantitative analysis in islet show intense nuclear expression and larger positive area for 8-OHdG in tacrolimus (TAC) groups. These changes are markedly decreased in TAC plus MK-0626 (M)-treated groups. (C) The TAC-induced serum 8-OHdG level is lowered by the addition of MK-0626. Immunoblot analysis of manganese superoxide dismutase (MnSOD) (D) and heme oxygenase-1 (HO-1, inducible form) and HO-2 (constitutive form) (E) in isolated pancreatic islets from the experimental rats. Note that the level of MnSOD and HO-1/HO-2 is lower in the TAC group; however, MK-0626 treatment leads to the recovery of its expression. Original magnifications, x400. <i>n</i> = 8 rats per group. ^#P<0.05 vs. control. ^$P<0.05 vs. TAC.</p

Effect of MK-0626 on the expression of GLP-1 receptor during tacrolimus-induced pancreatic islet injury.

<p>Immunohistochemistry (A) and quantitative analysis (B) of GLP-1 receptor (GLP-1R) in islets from tissue sections. Arrows indicate GLP-1R positive cells. (C) Immunoblot analysis of GLP-1R in isolated pancreatic islet from experimental groups. Note that the number and level of GLP-1R is lower in the tacrolimus (TAC) group, but MK-0626 (M)-treated groups exhibit a recovery of its expression compared with the TAC group. Original magnifications, x1000. <i>n</i> = 8 rats per group. ^#P<0.05 vs. control. ^$P<0.05 vs. TAC.</p