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AB151. Human tissue kallikrein 1 restores erectile function of streptozotocin-induced diabetic rats by activating PI3K/eNOS pathway and inhibiting oxidative stress

OBJECTIVE: Our previous study has proved the protective effect of human tissue kallikrein 1 (hKLK1) on erectile function of aged rats with the transgenic Sprague-Dawley (SD) rats harboring hKLK1 gene (TGR), but its role on diabetic erectile dysfunction (ED) is still unknown. METHODS: An 8-week-old m...

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Autores principales: Luan, Yang, Wang, Tao, Ruan, Yajing, Yang, Jun, Chen, Ruibao, Wang, Shaogang, Liu, Jihong, Ye, Zhangqun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: AME Publishing Company 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4708763/
http://dx.doi.org/10.3978/j.issn.2223-4683.2015.s151
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author Luan, Yang
Wang, Tao
Ruan, Yajing
Yang, Jun
Chen, Ruibao
Wang, Shaogang
Liu, Jihong
Ye, Zhangqun
author_facet Luan, Yang
Wang, Tao
Ruan, Yajing
Yang, Jun
Chen, Ruibao
Wang, Shaogang
Liu, Jihong
Ye, Zhangqun
author_sort Luan, Yang
collection PubMed
description OBJECTIVE: Our previous study has proved the protective effect of human tissue kallikrein 1 (hKLK1) on erectile function of aged rats with the transgenic Sprague-Dawley (SD) rats harboring hKLK1 gene (TGR), but its role on diabetic erectile dysfunction (ED) is still unknown. METHODS: An 8-week-old male wild type SD rats (WTR) and age matched TGR were randomly divided into five groups (n=8 in each group): (I) untreated WTR as the control; (II) untreated TGR; streptozotocin-induced diabetic (III) wild type rats (WTDM); and (IV) transgenic rats (TGDM); (V) TGDM with bradykinin receptor 2 inhibitor HOE140 (TGDMH). Erectile response, represented with the ratio of peak intracavernous pressure to systemic mean arterial pressure and area under the curve, were measured by cavernous nerve electrostimulation after 12 weeks. Penile corpus cavernosum (CC) tissues from all rats were harvested for hKLK1 gene identification and mechanism exploration. The related genes expressions of PI3 kinase (PI3K)/endothelial nitric oxide synthase (eNOS) pathway in CC of each group were assayed to detect the effect of hKLK1 in vivo. CC smooth muscle cell (CCSMC) isolated from TGR was administrated with pathway inhibitors and the intracellular concentration of cGMP and Ca(2+) were measured to confirm the potential signaling in vitro. In addition, the level of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, malondialdehyde (MDA) and activity of superoxide dismutase (SOD) were tested to indicate the local oxidative stress level in penile tissues from all groups. We also examined the reactive oxygen species (ROS) formation in CCSMC under the stimulation of high glucose to evaluate the anti-oxidative stress capacity of hKLK1 in vitro. RESULTS: hKLK1 attenuated the decrease of erectile function of WTDM while the protective effect was abolished in TGDMH. The decreased protein expression of PI3K (p55), AKT, eNOS, phospho-Enos (Ser1177), cyclic guanosine monophosphate (cGMP) and increased level of phospho-eNOS (Thr495), phosphodiesterase type 5 (PDE5), L-type calcium channel (ICa-L) in CC of WTDM were improved in that of TGDM, but deteriorated again with HOE140 in TGDMH. Meanwhile, hKLK1 could elevate the level of cGMP and reduce the concentration of Ca(2+) in CCSMC in vitro, but these changes were minified by inhibiting B2R, PI3K or eNOS. In addition, the CC of WTDM had higher protein expression of NADPH oxidase (p22phox, p47phox and NOX2), MDA level and lower activity of SOD when compared to WTR. However, hKLK1 improved these oxidative stress related parameters in TGDM and inhibition with HOE140 in TGDMH would abolish the advantages existed in TGDM. Meanwhile, the level of ROS elevated in high glucose treated CCSMC in a concentration-dependent manner and hKLK1 could considerably inhibit the generation of ROS in vitro. CONCLUSIONS: These results indicate that hKLK1 could restore diabetes-induced ED through activation of PI3K/eNOS signaling and inhibition of oxidative stress in the CC in vivo and in vitro.
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spelling pubmed-47087632016-01-26 AB151. Human tissue kallikrein 1 restores erectile function of streptozotocin-induced diabetic rats by activating PI3K/eNOS pathway and inhibiting oxidative stress Luan, Yang Wang, Tao Ruan, Yajing Yang, Jun Chen, Ruibao Wang, Shaogang Liu, Jihong Ye, Zhangqun Transl Androl Urol Moderated Poster Presentation OBJECTIVE: Our previous study has proved the protective effect of human tissue kallikrein 1 (hKLK1) on erectile function of aged rats with the transgenic Sprague-Dawley (SD) rats harboring hKLK1 gene (TGR), but its role on diabetic erectile dysfunction (ED) is still unknown. METHODS: An 8-week-old male wild type SD rats (WTR) and age matched TGR were randomly divided into five groups (n=8 in each group): (I) untreated WTR as the control; (II) untreated TGR; streptozotocin-induced diabetic (III) wild type rats (WTDM); and (IV) transgenic rats (TGDM); (V) TGDM with bradykinin receptor 2 inhibitor HOE140 (TGDMH). Erectile response, represented with the ratio of peak intracavernous pressure to systemic mean arterial pressure and area under the curve, were measured by cavernous nerve electrostimulation after 12 weeks. Penile corpus cavernosum (CC) tissues from all rats were harvested for hKLK1 gene identification and mechanism exploration. The related genes expressions of PI3 kinase (PI3K)/endothelial nitric oxide synthase (eNOS) pathway in CC of each group were assayed to detect the effect of hKLK1 in vivo. CC smooth muscle cell (CCSMC) isolated from TGR was administrated with pathway inhibitors and the intracellular concentration of cGMP and Ca(2+) were measured to confirm the potential signaling in vitro. In addition, the level of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, malondialdehyde (MDA) and activity of superoxide dismutase (SOD) were tested to indicate the local oxidative stress level in penile tissues from all groups. We also examined the reactive oxygen species (ROS) formation in CCSMC under the stimulation of high glucose to evaluate the anti-oxidative stress capacity of hKLK1 in vitro. RESULTS: hKLK1 attenuated the decrease of erectile function of WTDM while the protective effect was abolished in TGDMH. The decreased protein expression of PI3K (p55), AKT, eNOS, phospho-Enos (Ser1177), cyclic guanosine monophosphate (cGMP) and increased level of phospho-eNOS (Thr495), phosphodiesterase type 5 (PDE5), L-type calcium channel (ICa-L) in CC of WTDM were improved in that of TGDM, but deteriorated again with HOE140 in TGDMH. Meanwhile, hKLK1 could elevate the level of cGMP and reduce the concentration of Ca(2+) in CCSMC in vitro, but these changes were minified by inhibiting B2R, PI3K or eNOS. In addition, the CC of WTDM had higher protein expression of NADPH oxidase (p22phox, p47phox and NOX2), MDA level and lower activity of SOD when compared to WTR. However, hKLK1 improved these oxidative stress related parameters in TGDM and inhibition with HOE140 in TGDMH would abolish the advantages existed in TGDM. Meanwhile, the level of ROS elevated in high glucose treated CCSMC in a concentration-dependent manner and hKLK1 could considerably inhibit the generation of ROS in vitro. CONCLUSIONS: These results indicate that hKLK1 could restore diabetes-induced ED through activation of PI3K/eNOS signaling and inhibition of oxidative stress in the CC in vivo and in vitro. AME Publishing Company 2015-08 /pmc/articles/PMC4708763/ http://dx.doi.org/10.3978/j.issn.2223-4683.2015.s151 Text en 2015 Translational Andrology and Urology. All rights reserved.
spellingShingle Moderated Poster Presentation
Luan, Yang
Wang, Tao
Ruan, Yajing
Yang, Jun
Chen, Ruibao
Wang, Shaogang
Liu, Jihong
Ye, Zhangqun
AB151. Human tissue kallikrein 1 restores erectile function of streptozotocin-induced diabetic rats by activating PI3K/eNOS pathway and inhibiting oxidative stress
title AB151. Human tissue kallikrein 1 restores erectile function of streptozotocin-induced diabetic rats by activating PI3K/eNOS pathway and inhibiting oxidative stress
title_full AB151. Human tissue kallikrein 1 restores erectile function of streptozotocin-induced diabetic rats by activating PI3K/eNOS pathway and inhibiting oxidative stress
title_fullStr AB151. Human tissue kallikrein 1 restores erectile function of streptozotocin-induced diabetic rats by activating PI3K/eNOS pathway and inhibiting oxidative stress
title_full_unstemmed AB151. Human tissue kallikrein 1 restores erectile function of streptozotocin-induced diabetic rats by activating PI3K/eNOS pathway and inhibiting oxidative stress
title_short AB151. Human tissue kallikrein 1 restores erectile function of streptozotocin-induced diabetic rats by activating PI3K/eNOS pathway and inhibiting oxidative stress
title_sort ab151. human tissue kallikrein 1 restores erectile function of streptozotocin-induced diabetic rats by activating pi3k/enos pathway and inhibiting oxidative stress
topic Moderated Poster Presentation
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4708763/
http://dx.doi.org/10.3978/j.issn.2223-4683.2015.s151
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