[Preventive and therapeutic effects of Icariside â ¡ on radiation injury-induced erectile dysfunction in rats].

OBJECTIVE: To explore the preventive and therapeutic effects of Icariside Ⅱ (ICAⅡ) on radiation injury-induced ED (Ri-ED) in rats. METHODS: Twenty-four 10-week-old male SD rats received exposure of the prostate to single X-ray irradiation of 20 Gy, and were randomly equally divided into an Ri-ED group (6 survived and 6 died) and a treatment group treated with ICAⅡ at 4.5 mg/kg/d (9 survived and 3 died). Another 6 SD rats were taken as negative controls. After 4 weeks of continuous intragastric administration and 2 weeks of drug elution, the erectile function of the rats was evaluated by measurement of the maximum intracavernous pressure / mean arterial pressure (ICPmax/MAP), and the penile tissues were subjected to immunofluorescence staining, immunohistochemistry, Masson's trichrome staining, Western blot and detection of oxidative stress indicators. RESULTS: Compared with the negative controls, the rats in the Ri-ED group showed significant decreases in ICPmax/MAP (0.76 ± 0.09 vs 0.42 ± 0.06, P < 0.01), the number of nNOS-positive nerve fibers in the corpus cavernosum (10.17 ± 2.64 vs 3.17 ± 1.72, P < 0.01), the content of endothelial cells (1.39 ± 0.30 vs 0.35 ± 0.12, P < 0.01), the expressions of nNOS (0.42 ± 0.08 vs 0.08 ± 0.01, P < 0.01) and eNOS (0.99 ± 0.24 vs 0.12 ± 0.08, P < 0.01) and the activity of superoxide dismutase (SOD) (ï¼»343.73 ± 58.57ï¼½ vs ï¼»153.50 ± 34.06ï¼½ U/mg prot, P < 0.01), but an increase in the malondialdehyde (MDA) level (ï¼»1.80 ± 0.31ï¼½ vs ï¼»3.25 ± 0.21ï¼½ nmol/mg prot, P < 0.01). In comparison with the Ri-ED group, the animals treated with ICAⅡ exhibited remarkably increased ICP/MAP (0.42 ± 0.06 vs 0.66 ± 0.07, P < 0.01), number of nNOS-positive nerve fibers (3.17 ± 1.72 vs 7.33 ± 1.75, P < 0.05), content of endothelial cells (0.35 ± 0.12 vs 1.07 ± 0.36, P < 0.01), and expressions of nNOS (0.08 ± 0.01 vs 0.16 ± 0.05, P < 0.05) and eNOS (0.12 ± 0.08 vs 0.86 ± 0.30, P < 0.01) in the corpus cavernosum, but a decreased level of MDA (ï¼»3.25 ± 0.21ï¼½ vs ï¼»2.17 ± 0.55ï¼½ nmol/mg prot, P < 0.05). In addition, ICAⅡ effectively reduced radiation injury-induced mortality of the rats. CONCLUSION: IcarisideⅡ can significantly improve ED and pathological changes and reduce mortality caused by radiation injury in rats, which may be related to its ability of improving radiation-induced oxidative stress.

Therapeutic effects of adipose-derived stem cells-based microtissues on erectile dysfunction in streptozotocin-induced diabetic rats.

This study aimed to explore the therapeutic effects of adipose-derived stem cells (ADSCs)-based microtissues (MTs) on erectile dysfunction (ED) in streptozotocin (STZ)-induced diabetic rats. Fifty-six 8-week-old Sprague-Dawley rats received intraperitoneal injection of STZ (60 mg kg-1 ), and 8 weeks later, the determined diabetic rats randomly received intracavernous (IC) injection of phosphate buffer solution (PBS), ADSCs, or MTs. Another eight normal rats equally got IC injection of PBS. MTs were generated with a hanging drop method, and the injected cells were tracked in ADSC- and MT-injected rats. Four weeks after the treatments, intracavernous pressure (ICP), histopathological changes in corpus cavernosum (CC), and functional proteins were measured. Rat cytokine antibody array was used to detect ADSCs or MTs lysate. The results showed that MTs expressed vascular endothelial growth factor (VEGF), nerve growth factor (NGF), and tumor necrosis factor-stimulated gene-6 (TSG-6). MTs injection had a higher retention than ADSCs injection and MTs treatment improved ICP, neuronal nitric oxide synthase (nNOS) expression, smooth muscle, and endothelial contents in diabetic rats, ameliorated local inflammation in CC better. Thus, our findings demonstrate that IC injection of MTs improves erectile function and histopathological changes in STZ-induced diabetic rats and appears to be more promising than traditional ADSCs. The underlying mechanisms involve increased cell retention accompanied with neuroprotection and anti-inflammatory behaviors of the paracrine factors.

Conversion of adipose-derived stem cells into natural killer-like cells with anti-tumor activities in nude mice.

Efforts to develop peripheral blood-derived nature killer (NK) cells into therapeutic products have been hampered by these cells' low abundance and histoincompatibility. On the other hand, derivation of NK-like cells from more abundant cell sources such as embryonic stem cells (ESCs) and umbilical cord blood (UCB) requires the selection of rare CD34+ cells. Thus, we sought to convert adipose-derived stem cells (ADSCs), which are abundant and natively CD34+, into NK-like cells. When grown in hematopoietic induction medium, ADSCs formed sphere clusters and expressed hematopoietic markers CD34, CD45, and KDR. Further induction in NK cell-specific medium resulted in a population of cells that expressed NK cell marker CD56, and thus termed ADSC-NK. Alternatively, the hematopoietically induced ADSCs were transduced with NK cell-specific transcription factor E4BP4 prior to induction in NK cell-specific medium. This latter population of cells, termed ADSC-NKE, expressed CD56 and additional NK cell markers such as CD16, CD94, CD158, CD314, FasL, and NKp46. ADSC-NKE was as potent as NK leukemia cell NKL in killing breast cancer cell MCF7 and prostate cancer cells DU145, PC3, LnCap, DuPro, C4-2 and CWR22, but exhibited no killing activity toward normal endothelial and smooth muscle cells. In nude mice test ADSC-NKE was able to significantly delay the progression of tumors formed by MCF7 and PC3. When injected into immunocompetent rats, ADSC-NKE was detectable in bone marrow and spleen for at least 5 weeks. Together, these results suggest that ADSCs can be converted into NK-like cells with anti-tumor activities.

Evaluation of the effect of different doses of low energy shock wave therapy on the erectile function of streptozotocin (STZ)-induced diabetic rats.

To investigate the therapeutic effect of different doses of low energy shock wave therapy (LESWT) on the erectile dysfunction (ED) in streptozotocin (STZ) induced diabetic rats. SD rats (n = 75) were randomly divided into 5 groups (normal control, diabetic control, 3 different dose LESWT treated diabetic groups). Diabetic rats were induced by intra-peritoneal injection of STZ (60 mg/kg) and rats with fasting blood glucose ≥ 300 mg/dL were selected as diabetic models. Twelve weeks later, different doses of LESWT (100, 200 and 300 shocks each time) treatment on penises were used to treat ED (7.33 MPa, 2 shocks/s) three times a week for two weeks. The erectile function was evaluated by intracavernous pressure (ICP) after 1 week washout period. Then the penises were harvested for histological study. The results showed LESWT could significantly improve the erectile function of diabetic rats, increase smooth muscle and endothelial contents, up-regulate the expression of α-SMA, vWF, nNOS and VEGF, and down- regulate the expression of RAGE in corpus cavernosum. The therapeutic effect might relate to treatment dose positively, and the maximal therapeutic effect was noted in the LESWT300 group. Consequently, 300 shocks each time might be the ideal LESWT dose for diabetic ED treatment.

Transdifferentiation of human adipose-derived stem cells into urothelial cells: potential for urinary tract tissue engineering.

Autologous urothelial cells (UCs) provide a cell source for urinary tissue engineering because they can be used safely due to their lack of immunogenicity. However, these cells cannot be harvested under the following circumstances: malignancy, infection and organ loss, etc. Human adipose-derived stem cells (HADSCs) possess the traits of high differentiation potential and ease of isolation, representing a promising resource for tissue engineering and regenerative medicine. Nevertheless, HADSCs have been poorly investigated in urology and the optimal approaches to induce HADSCs into urothelium are still under investigation. In this study, we hypothesized that the change of microenvironment by a conditioned medium was essential for the transdifferentiation of HADSCs into UCs. We then used a conditioned medium derived from urothelium to alternate the microenvironment of HADSCs. After 14 days of culture in a conditioned medium, about 25-50% HADSCs changed their morphology into polygonal epithelium-like shapes. In addition, these cells expressed up-regulating of urothelial lineage-specific markers (uroplakin 2and cytokeratin-18) and down-regulating of mesenchymal marker (vimentin) in RNA and protein level, respectively, which confirmed that HADSCs were induced into urothelial lineage cells. We also measured the growth factors in the conditioned medium in order to analyze the molecular mechanisms regulating transdifferentiation. We observed that the expression levels of PDGF-BB and VEGF were significantly higher than those of the control group after 14 days induction, suggesting they were abundantly secreted into the medium during the culturing period. In conclusion, HADSCs showed in vitro the upregulation of markers for differentiation towards urothelial cells by culturing in an urothelial-conditioned medium, which provides an alternative cell source for potential use in urinary tract tissue engineering.