Activation of VEGF and ERK1/2 and improvement of urethral function by adipose-derived stem cells in a rat stress urinary incontinence model.

OBJECTIVE: To investigate the injected autologous adipose-derived stem cells (ADSCs) in improving stress urinary incontinence in a rodent model of parturition-related stress incontinence and the possible mechanism. METHODS: The 40 rats were developed stress urinary incontinence models by postpartum balloon dilation of the vagina for 4 hours followed by bilateral ovariectomy. ADSCs were isolated from the peri-ovarian fat and labeled with thymidine analog 5-ethynyl-2-deoxyuridine (EdU). Twenty stress urinary incontinence rats received peri-urethral injection of phosphate-buffered saline as the negative controls and the other 20 stress urinary incontinence rats received peri-urethral injection of EdU-labeled ADSCc. Twenty control rats underwent sham ovariectomy without balloon dilation and served as positive controls. Four weeks later, voiding function was assessed by cystometry. Urethral histologic examination (Masson trichrome stain, picrosirius red stain, Hart elastin stain, Gordon and Sweet stain, and immunohistochemical stain) and Western blot were performed on urethral tissues. RESULTS: Both leak point pressure and bladder capacity were significantly increased in ADSC-treated rats, compared to the balloon-injured ovariectomized rats. Histologic examination revealed normalized appearance of the fibromuscular structure of the urethra as well as increased peri-urethral blood vessel density in ADSC-treated rats. On Western blot, vascular endothelial growth factor and P-extracellular signal-regulated kinases (ERKs)1/2 protein was expressed at a higher rate in tissues from ADSC-treated rats compared to phosphate-buffered saline-treated rats. CONCLUSION: Peri-urethral injection of ADSCs is associated with more normal urinary function and urethral structure in rats with parturition-related incontinence. The activation of vascular endothelial growth factor and ERK1/2 may be responsible for the paracrine effects from ADSCs.

Effects of icariside II on improving erectile function in rats with streptozotocin-induced diabetes.

Icariin and icariside II (ICA II), 2 active components isolated from herba epimedii, have a closely structural relationship. There is evidence that icariin may be useful in the treatment of erectile dysfunction (ED); however, the study on the therapeutic efficacy of ICA II on ED is currently scant. We investigated the effects of ICA II on improving erectile function of rats with streptozocin-induced diabetes. Fifty 8-week-old Sprague-Dawley rats were randomly distributed into normal control and diabetic groups. Diabetes was induced by a one-time intraperitoneal injection of streptozocin (60 mg/kg). Three days later, the diabetic rats were randomly divided into 4 groups including a saline-treated placebo arm and 3 ICA II-treated models (1, 5, and 10 mg/kg/d). After 3 months, penile hemodynamics was measured by cavernous nerve electrostimulation (CNE) with real time intracorporal pressure assessment. Penises were harvested with subsequent histological examination (picrosirius red stain, Hart elastin stain, and immunohistochemical stain) and Western blots to explore the expression of the nitric oxide-cyclic guanosine monophosphate (NO-cGMP) and transforming growth factor ß1 (TGFß1)/Smad2 signaling pathways. Diabetes significantly attenuated erectile responses to CNE. Diabetic rats had decreased corpus cavernosum smooth muscle/collagen ratio and endothelial cell content relative to the control group. The ratio of collagen I to III was significantly lower in the corpora of diabetic rats; furthermore, cavernous elastic fibers were fragmented in the diabetic animals. Neuronal nitric oxide synthase (nNOS), endothelial nitric oxide synthase, and vascular endothelial growth factor were expressed at lower levels in the diabetic group; ICA II-treated diabetic rats had higher expression in the penis relative to placebo-treated diabetic animals. Both the TGFß1/Smad2/connective tissue growth factor (CTGF) signaling pathway and apoptosis were down-regulated in the penis from ICA II-treated rats. ICA II treatment attenuates diabetes-related impairment of penile hemodynamics, likely by increasing smooth muscle, endothelial function, and nNOS expression. ICA II could alter corpus cavernosum fibrous-muscular pathological structure in diabetic rats, which could be regulated by the TGFß1/Smad2/CTGF and NO-cGMP signaling pathways.

[Effects of icariin and icariside II on eNOS expression and NOS activity in porcine aorta endothelial cells].

OBJECTIVE: To investigate the effects of icariin and icariside II on eNOS expression and NOS activity in endothelial cells and possible mechanisms using EGFR over-expressed porcine aorta endothelial (PAE) cell line. METHODS: The EGFR gene was transfected into PAE cells and genetic stable cell line (PAE-EGFR) was selected. 12.5 µmol/L of icariin and of icariside II were used to treat the PAE and PAE-EGFR cells respectively for 48 h, the eNOS expression in each group was observed. EGF was also used to treat the cells to observe the regulatory effects of icariin and icariside II on NOS activity. The regulatory effects of icariin and icariside II on NOS activity were also observed, and sildenafil was used as a control. RESULTS: Western blot showed that the basic value of eNOS expression was higher in PAE-EGFR group compared with that in PAE group, both of icariin and icariside II increased the eNOS expression in PAE and PAE-EGFR group (P<0.01), and the value of eNOS expression was higher in PAE-EGFR group than that in PAE group. In the PAE-EGFR cell line, the NOS activity reached (15.37 ± 1.49) u/mg when the concentration of icariside II was 10(-8) mol/L, which was 4.66 u/mg more than that in the PAE cell line. When the concentration reached 10(-7), 10(-6) or 10(-5) mol/L, the change of NOS activity in PAE-EGFR group was greater than that in PAE group (P<0.01). icariin also increased the NOS activity in PAE and PAE-EGFR cells, but the activity was 20% lower compared with icariside II group, however, Sildenafil showed no influence on NOS activity. CONCLUSION: Icariin and icariside II may increase the eNOS expression through activating EGF-EGFR pathway in PAE cell, by which endothelial cells function could be regulated and the better effect was noted in icariside II compared to icariin.

[Corpus cavernosum-corpus spongiosum shunt plus intracavernous tunneling for the treatment of prolonged ischemic priapism].

OBJECTIVE: To investigate the efficacy and safety of corpus cavernosum-corpus spongiosum shunt (CC-CSS) plus intracavernous tunneling(CC-CSS+ICT) for the treatment of prolonged ischemic priapism (PIP) were investigated. METHODS: Of 21 patients with PIP, 11 (Group A) underwent CC-CSS and 10 (Group B) CC-CSS+ICT surgery. The penile hardness score (PHS) and pain visual analogue score (PVAS) were used to assess the efficacy of the surgery. RESULTS: The erectile functions of the two groups were normal (IIEF5 23.6+/-1.1) before the onset of PIP, and the duration of PIP was (3.4+/-1.3) d. PHS 3.9+/-0.4, and PVAS 8.4+/-0.7. There was no statistical difference between the two groups (P>0.05). On 1, 3 and 5 days after the operation, the PHS and PVAS of Group B decreased significantly than those of Group A (P<0.05). CONCLUSION: CC-CSS+ICT could quickly restore penile detumescence and relieve pain as compared with CC-CSS, which might be a safe and effective method for the treatment of PIP.

[Preliminary research on the relationship between inhibitory effect of triamcinolone acetonide on hypertrophic scar and oxygen free radicals in rabbits].

OBJECTIVE: To investigate the relationship between inhibitory effect of triamcinolone acetonide(TA) on hypertrophic scar and oxygen free radicals in rabbits. METHODS: 18 New Zealand rabbits were used. 14 rabbits were selected randomly to construct animal models of hypertrophic scar on the ears. Another 4 rabbits (8 ears) were used as controls. 6 weeks after operation, the hypertrophic scar on the ears were randomly divided to receive intra-lesion injection of TA (n=10), or normal saline (n=10), or nothing (n=8, sham group). 9 weeks after operation, scar specimens were taken for scar thickness measurement, fibroblast counting under microscopy and MDA content detection by spectrophotometric method. RESULTS: (1) 3 weeks after TA treatment, the scar became very thin and soft with a similar color to normal skin and a smooth surface; (2) Histologic study showed the collagen fibers in TA group were reduced markedly and arranged parallelly; (3) Compared with normal skin, the fibroblast density in sham and saline groups increased significantly (P < 0.05), while it was not markedly different in TA group (P > 0.05). There was also no significant difference in scar hypertrophic index between sham and saline groups (P > 0.05), but the scar hypertrophic index was decreased dramatically in TA group (P < 0.05); (4) The MDA content was highest in TA group (P < 0.05), followed by that in sham and saline groups (P < 0.05), while there was no difference between these two groups (P > 0.05). It was lowest in normal control group (P < 0.05). CONCLUSIONS: The oxygen free radicals in the hypertrophic scar can be further increased by local injection of TA.