Zhe Tang

Lipoxin A4 improves erectile dysfunction in rats with type I diabetes by inhibiting oxidative stress and corporal fibrosis.

Previous studies have shown that oxidative stress and corporal fibrosis in penile tissues of rats were key pathological factors of erectile dysfunction induced by diabetic mellitus (DMED). Lipoxin A4 (LXA4) was reported to inhibit oxidative stress and fibrosis diseases, while whether it could exert a protective role on erectile function was not clear. Type I diabetic mellitus (DM) was induced in thirty male 10-week-old Sprague-Dawley rats using streptozotocin. Ten weeks later, twenty-two rats with DMED confirmed by an apomorphine test were divided into two groups: the DMED group (n = 11) and the DMED + LXA4 group (n = 11; LXA4 injection daily for 4 weeks). In addition, another ten age-matched rats formed the Control group. We found that erectile function was significantly impaired in the DMED group compared with the Control group, but was improved in the DMED + LXA4 group. Similarly, the over-activated oxidative stress and impaired endothelial function in the DMED group were both improved in the DMED + LXA4 group. Moreover, the DMED group showed serious corporal fibrosis, which was also inhibited by the treatment of LXA4 in the DMED + LXA4 group. Taken together, LXA4 could exert an inhibition role on oxidative stress and fibrosis to improve DMED effectively.

MP43-18 HUMAN TISSUE KALLIKREIN 1 RESCUES ERECTILE FUNCTION IN RATS WITH HYPERHOMOCYSTEINAEMIA BY PROTECTING ENDOTHELIAL FUNCTION AND INHIBITING FIBROSIS

INTRODUCTION AND OBJECTIVES: To investigate the detailed mechanism of erectile dysfunction (ED) induced by hyperhomocysteinaemia (HHcy) in rats and determine whether the Human Tissue Kallikrein 1 (hKLK1) might improve it, as we have proved the protective role of hKLK1 on erectile function in aged rats. METHODS: We established a rat model of HHCy through dietary-rich methionine (Met) in male Sprague-Dawley (SD) rats. Male wild-type SD rats (WTR) and transgenic rats harboring the hKLK1 gene (TGR) were fed to 10 weeks of age. Then 24 WTRs were divided into control (n1⁄48), the low-dose (4% Met, n1⁄48), and the highdose (7% Met, n1⁄48). Another 8 age-matched TGRs with the highdose formed the TGR+7%Met group. 30 days later, erectile function, level of total homocysteinaemia (tHcy), oxidative stress, endothelial function, cavernous nerve function and fibrosis of all groups were determined. RESULTS: hKLK1 in the TGR+7%Met group could greatly decrease the tHcy levels and improve ED induced by HHcy in rats. For the endothelial function, hKLK1 could preserve the endothelial cell-cell junction, enhance endothelial regeneration and activated the Akt/eNOS signaling pathway. Together with the promotion of hKLK1 on nNOS expression, the NO/cGMP signaling pathway activity was also increased. For the fibrosis, hKLK1 could preserve normal corpus cavernosum structure through inhibiting apoptosis and promoting autophagy on corpus cavernosum smooth muscle cells. In addition, hKLK1 also inhibited the fibrosis-related signaling pathway activity. CONCLUSIONS: hKLK1 might effectively improve ED induced by HHcy in rats by protecting endothelial function, promoting cavernous nerve function and inhibiting fibrosis, which suggested hKLK1 might be a potential treatment method for ED.

Comparison of intracytoplasmic sperm injection outcome with fresh versus frozen-thawed testicular sperm in men with nonobstructive azoospermia: a systematic review and meta-analysis

PurposeThe purpose of the study is to explore testicular sperm cryopreservation in patients with nonobstructive azoospermia (NOA) whether affect the outcome of subsequent intracytoplasmic sperm injection (ICSI).MethodsA systematic review and meta-analysis was conducted by searching the MEDLINE and EMBASE databases for relevant published studies in English language (1997–2017). Studies were eligible if they included the comparison of using fresh and frozen-thawed testicular sperm followed by ICSI. Two reviewers independently performed data extraction, quality assessment and assessed the risk of bias. The overall summary risk estimated the number of events. A meta-analysis was conducted using a random effects or fixed effects model analysis according to the test of heterogeneity.ResultsA total of 17 studies with 1,261 ICSI cycles were identified. Analysis of the present data showed no difference in the fertilization outcome when comparing fresh versus frozen-thawed spermatozoa (RR = 1.02, 95% CI 0.86–1.09). Similarly, no difference in CR (RR = 1.01, 95% CI 0.96–1.05), good embryo rate (RR = 1.01, 95% CI 0.95–1.09), and IR (RR = 0.93, 95% CI 0.66–1.30) was observed if the spermatozoa was fresh or frozen-thawed. Finally, no difference in CPR or LBR was noted when using fresh or frozen-thawed cycles were analyzed separately (RR = 1.03, 95% CI 0.86–1.24; RR 1.11, 95% CI 0.88–1.41, respectively).ConclusionsIn men with NOA, the ICSI outcome is not affected by whether the retrieved testicular sperm is fresh or frozen. Sperm cryopreservation ought to be considered in every surgical sperm retrieval case, which remain feasible even in patients with few testicular sperm retrieved.

AB088. FTY720 supplementation partially improves erectile dysfunction in rats with streptozotocin-induced type 1 diabetes through inhibition of endothelial dysfunction and corporal fibrosis

Background To investigate whether FTY720, approved in 2010 for the treatment of patients with the relapsing-remitting form of multiple sclerosis, could ameliorate erectile dysfunction induced by diabetes mellitus (DMED). Methods Thirty-two Sprague-Dawley rats (8 weeks old) were induced type I DM and the other eight rats formed the control (n=8). Eight weeks later, 17 rats with DMED tested with an apomorphine test were divided in two groups: DMED (n=8) and DMED + FTY720 (1 mg/kg/d; n=9). Treatment of FTY720 lasted for 4 weeks. Results Impaired erectile function, inhibited S1P3/Akt/NO/cGMP activity, serious corporal fibrosis and over-activated pathways (the Smad and non-Smad) were found in the DMED group compared with the control, while FTY720 partly but significantly improved these pathological changes induced by DM. Conclusions FTY720 supplementation inhibited endothelial dysfunction and corporal fibrosis, ultimately leading to partial improvement of DMED in rats. This finding provides evidence for a potential treatment method for DMED.

AB087. Preserved erectile function in the hyperhomocysteinaemia transgenic rats harboring human tissue kallikrein

Background To investigate the role of human tissue kallikrein 1 (hKLK1) gene on the erectile dysfunction (ED) of induced by hyperhomocysteinaemia (HHcy) in rats. Methods The HHcy rat model was formed by a methionine (Met)-rich diet in SD rats. Here, 32 rats, 10-week-age, were divided in four groups: control (n=8), low-dose (4% Met; n=8), high-dose (7% Met; n=8) and transgenic rats (TGR +7% Met; n=8). Thirty days later, erectile function and related targets were tested. Results ED, impaired endothelial and smooth muscle function, and pathological changes (a higher apoptosis level and a lower autophagy level) were showed in the 4% Met and 7% Met groups compared with the control, while were all markedly diminished by the hKLK1 gene in the TGR +7% Met group. Conclusions These data suggested that hKLK1 might play an inhibition role on HHcy-induced ED in rats by protection of endothelial function and inhibition of oxidative stress and corporal fibrosis.

MP81-14 HUMAN TISSUE KALLIKREIN 1 AMELIORATES ERECTILE FUNCTION VIA MODULATING AUTOPHAGY AND ACTIVATING HIF-1?/COX-2 PATHWAY IN AGED TRANSGENIC RATS

normalized to KCl). Data are presented as means SEM. Data were statistically analyzed using a one way ANOVA followed by Tukeys test(GraphPad Prism software). RESULTS: Vaginal wet weight was decreased (P<0.05) in OVXVV animals compared to SHVV, an effect reversed by local estrogen delivery. Qualitative analysis of vaginal cross sections indicated reversal of ovariectomy induced atrophy of the vaginal muscularis with estrogen treatment. In vitro contractility studies with carbachol demonstrated a trend of a lower EC50 (increased sensitivity) of vaginal strips obtained from OVXVV animals compared to SHVV and OVXVE. The amplitude of contraction to 10 uM carbachol was greater of proximal strips from OVXVV animals compared to SHVV and OVXVE (P<0.05). CONCLUSIONS: Our results indicate that vaginal estrogen is effective at reversing not only OVX induced atrophy of the epithelium but also the vaginal muscularis. We report an increased contractile response to carbachol in OVXVV animals, an effect also reversed by vaginal estrogen. Interestingly, previous studies have shown an increase in vaginal sensory innervation in ovariectomized rodents. More studies are needed to evaluate changes in autonomic innervation with this animal model to identify new therapeutic uses of vaginal estrogen treatment.

Involvement of DDAH/ADMA/NOS/cGMP and COX-2/PTGIS/cAMP Pathways in Human Tissue Kallikrein 1 Protecting Erectile Function in Aged Rats

Our previous studies had reported that Human Tissue Kallikrein 1 (hKLK1) preserved erectile function in aged transgenic rats, while the detailed mechanism of hKLK1 protecting erectile function in aged rats through activation of cGMP and cAMP was not mentioned. To explore the latent mechanism, male wild-type Sprague-Dawley rats (WTR) and transgenic rats harboring the hKLK1 gene (TGR) were fed to 4 and 18 months old and divided into four groups: young WTR (yWTR) as the control, aged WTR (aWTR), aged TGR (aTGR) and aged TGRs with HOE140 (aTGRH). Erectile function of all rats was evaluated by cavernous nerve electrostimulation method and measured by the ratio of intracavernous pressure/ mean arterial pressure (ICP/MAP) in rats. Expression levels of cAMP and cGMP were assessed, and related signaling pathways were detected by western blot, immunohistochemistry and RT-PCR. Our experiment results showed erectile function of the aWTR group and aTGRH group was lower compared with those of other two groups. Also, expression levels of cAMP and cGMP were significantly lower than those of other two groups. Moreover, expressions of related signaling pathways including DDAH/ADMA/NOS/cGMP and COX-2/PTGIS/cAMP were also downregulated in the corpus cavernosum of rats in aWTR group. Our finding revealed hKLK1 played a protective role in age-related ED. The DDAH/ADMA/NOS/cGMP and COX-2/PTGIS/cAMP pathways that were linked to the mechanism hKLK1 could increase the levels of cGMP and cAMP, which might provide novel therapy targets for age-related ED.