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

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Asian Journal of Andrology 2017年第1期19卷 91-97页

作者： Feng Zhou Hua xin Yong-De Xu Hong-En Lei Bi-cheng Yang Rui-Li Guan Meng Li Jian-Quan Hou zhong-cheng xin Molecular Biology Laboratory of Andrology Center Peking University First Hospital Peking University Beijing 100034 China Department of Urology First Affiliated Hospital of Soochow University Soochow University Suzhou 215006 China Department of Ophthalmology Beijing Chao-Yang Hospital Capital Medical University Beijing 100043 China Department of Urology General Hospital of Ningxia Medical University Ningxia Medical University Ningxia 750021 China

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.

关键词： adipose-derived stem cells diabetes erectile dysfunction microtissues paracrine factors

Comparative study of intracavernous pressure and cavernous pathology after bilateral caver nous n erve crushing and resection in rats

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Asian Journal of Andrology 2020年第6期22卷 629-635页

作者： Meng Li Yi-Ming Yuan Bi-cheng Yang Sheng-Ji Gu Hui-Xi Li zhong-cheng xin Dong Fang Rui-Li Guan Molecular Biology Laboratory of Andrology Center Peking University First HospitalPeking UniversityBeijing 100034China

This study aimed to compare the effects of bilateral cavernous nerve crushing(BCNC)and bilateral cavernous nerve resection(BCNR)on intracavernous pressure(ICP)and cavernous pathology in rats and to explore the optimal treatment time for the BCNC and BCNR ***-two male rats aged 12 weeks were randomly divided into three equal groups:Sham(both cavernous nerves exposed only),BCNC(BCN crushed for 2 min),and BCNR(5 mm of BCN resected).Erectile function was then measured at 1 week,3 weeks,and 5 weeks after nerve injury,and penile tissues were harvested for histological and molecular analyses by immunohistochemistry,immuno fluoresce nee,Western blot,and cytokine *** found that erectile function parameters including the maximum,area,and slope of ICP/mean arterial pressure(MAP)significantly decreased after BCNR and BCNC at 1 week and 3 *** 5 weeks,no significant differences were observed in ICP/MAP between the BCNC and Sham groups,whereas the ICP/MAP of the BCNR group remained significantly lower than that of the Sham *** BCNC and BCNR,the amount of neuronal-nitric oxide synthase-positive fibers,smooth muscle cells,and endothelial cells decreased,whereas the amount of collage n III con tent *** pathological cha nges recovered over time,especially in the BCNC *** fin dings demonstrate that BCNC leads to acute and reversible erectile dysfunction,thus treatme nt time should be restricted to the first 3 weeks *** contrast,the self-healing ability of the BCNR model is poor,making it more suitable for long-term treatment research.

关键词： caver nous nerve crushing cavernous nerve resection erectile dysfunction in tracavernous pressure

Effect of icarisid II on diabetic rats with erectile dysfunction and its potential mechanism via assessment of AGEs, autophagy, roTOR and the NO-cGMP pathway

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Asian Journal of Andrology 2013年第1期15卷 143-148页

作者： Jian Zhang Ai-Min Li Bao-xing Liu Fei Han Feng Liu Shao-Peng Sun xin Li Shu-Jin Cui Shao-zhong Xian Guang-Qi Kong zhong-cheng xin Zhi-Li Ji Department of Urology Capital Medical University Luhe Hospital Beijing 101149 China Andrology Center Peking University First Hospital Beijing 100034 China Department of Andrology China-Japan Friendship Hospital Beijing 100029 China

Erectile dysfunction （ED） is a major complication of diabetes mellitus. Icariin has been shown to enhance erectile function through its bioactive form, icarisid Ih This study investigates the effects of icarisid Ⅱ on diabetic rats with ED and its potential mechanism viathe assessment of advanced glycosylation end products （AGEs）, autophagy, mTOR and the NO-cGMP pathway. Icarisid Ⅱ was extracted from icariin by an enzymatic method. In the control and diabetic ED groups, rats were administered normal saline; in the icarisid Ⅱ group, rats were administered icarisid Ⅱ intragastrically. Erectile function was evaluated by measuring intracavernosal pressure/mean arterial pressure （ICP/MAP）. AGE concentrations, nitric oxide synthase （NOS） activity and cGMP concentration were assessed by enzyme immunoassay. Cell proliferation was analysed using methyl thiazolyl tetrazolium assay and flow cytometry. Autophagosomes were observed by transmission electron microscopy, monodansylcadaverine staining and GFP-LC3 Iocalisation. The expression of NOS isoforms and key proteins in autophagy were examined by western blot. Our results have shown that Icarisid Ⅱ increased ICP/MAP values, the smooth muscle cell （SMC） growth curve, S phase and SMC/collagen fibril （SMC/CF） proportions and decreased Beclin 1 （P〈0.05）. Icarisid Ⅱ significantly increased the proliferative index and p-p70S6K（Thr389） levels and decreased the numbers of autophagosomes and the levels of LC3-11 （P〈0.01）. Icarisid Ⅱ decreased AGE concentrations and increased cGMP concentration, NOS activity （P〈0.05） and cNOS levels （P〈0.01） in the diabetic ED group. Therefore, Icarisid Ⅱ constitutes a promising compound for diabetic ED and might be involved in the upregulation of SMC proliferation and the NO-cGMP pathway and the downregulation of AGEs, autophagy and the mTOR pathway.

关键词： advanced glycosylation end products (AGEs) autophagy cell proliferation diabetes mellitus (DM) erectile dysfunction (ED) icariin icarisid II mTOR NO-cGMP NOS activity

Effect of tankyrase 1 on autophagy in the corpus cavernosum smooth muscle cells from ageing rats with erectile dysfunction and its potential mechanism

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Asian Journal of Andrology 2010年第5期12卷 744-752页

作者： Jian Zhang Xiao-Jun Wu De-Xiang Zhuo Tao Liu Wei-Ren Li Ze-Bin Mao zhong-cheng xin Andrology Center First Hospital Peking University Beijing 100009 China Institute of Urinary Surgery Southwest Hospital Third Military Medical University Chongqing 400038 China The Department of Biochemistry and Molecular Biology Health Science Center Peking University Beijing 100191 China

This study compared tankyrase 1 expression and autophagy quantity between erectile dysfunction （ED） and non-ED rats＇ corpus cavernosum smooth muscle cells （CSMCs）. This study aslo explored the effect and possible mechanism of tankyrase 1 on autophagy and cell proliferation in ageing ED rats＇ CSMCs. The intracavernous pres- sure and mean systemic arterial pressure were measured to investigate erectile function so that eight 24-month-old ED and eight 8-month-old male Wistar rats were choosed respectively. The rat CSMCs were isolated and cultured by enzyme digestion, in which tankyrase 1 expression and autophagy quantity were compared. Tankyrase 1 over-expression was induced with plasmid transfection by Lipofectamine^TM. The effect of tankyrase 1 overexpression on proliferation, autophagy and mTOR pathway in 24-month-old ED rats＇ CSMCs was measured by the cell growth curve in MTT assay, cell cycle analysis in flow cytometry （FCM）, key protein expression in Western blot, autophagy quantity in transmission electron microscopy, monodansylcadaverine staining and GFP-LC3 fluorescence. The primary CSMCs were confirmed by immunofluorescence, and the purity was 99.1% in FCM. Compared with that of 8-month-old rats, tankyrase 1 expression and autophagy quantity significantly decreased in 24-month-old ED rats＇ primary CSMCs （P 〈 0.01）. Tankyrase 1 overexpression significantly increased the growth rate （P 〈 0.05） and increased the S phase of cell cycle （P 〈 0.01）. The autophagosome quantity was remarkably increased （P 〈 0.01）, LC3-Ⅰ/Ⅱ and Beclin 1 were upregulated （P 〈 0.01 and P 〈 0.05）, and p-p70S6K （Thr389） was downregulated in 24-month-old ED rat CSMCs （P 〈 0.05）. In conclusion, Tankyrase 1 and autophagy decrease in the CSMCs from aging rats with ED, and tankyrase 1 may have a positive effect on proliferation by enhancing autophagy and regulating the mTOR signalling pathway.

关键词： ageing autophagy corpora cavernosum corpus cavemosum smooth muscle cells erectile dysfunction senile tankyrase

Autophagic deficiency is related to steroidogenic decline in aged rat Leydig cells

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Asian Journal of Andrology 2011年第6期13卷 881-888页

作者： Wei-Ren Li Liang Chen Zhi-Jie Chang Hua xin Tao Liu Yan-Quan Zhang Guang-Yong Li Feng Zhou Yan-Qing Gong Zhe-Zhu Gao zhong-cheng xin Andrology Center Peking University First Hospital Peking University Beijing 100034 China Department of Urology China Meitan General Hospital Beijing 100028 China Department of Biological Sciences and Biotechnology School of Medicine Tsinghua University Beijing 100084 China

Late-onset hypogonadism （LOH） is closely related to secondary androgen deficiency in aged males, but the mechanism remains unclear. In this study, we found that reduced testosterone production in aged rat Leydig cells is associated with decreased autophagic activity. Primary rat Leydig cells and the TM3 mouse Leydig cell line were used to study the effect of autophagic deficiency on Leydig cell testosterone production. In Leydig cells from young and aged rats, treatment with wortmannin, an autophagy inhibitor, inhibited luteinising hormone （LH）-stimulated steroidogenic acute regulatory （STAR） protein expression and decreased testosterone production. In contrast, treatment with rapamycin, an autophagy activator, enhanced LH-stimulated steroidogenesis in Leydig cells from aged, but not young, rats. Intracellular reactive oxygen species （ROS） levels were increased in both young and aged Leydig cells treated with wortmannin but decreased only in aged Leydig cells treated with rapamycin. Furthermore, an increased level of ROS, induced by H2O2, resulted in LH-stimulated steroidogenic inhibition. Finally, knockdown of Beclin 1 decreased LH-stimulated StAR expression and testosterone production in TM3 mouse Leydig cells, which were associated with increased intracellular ROS level. These results suggested that autophagic deficiency is related to steroidogenic decline in aged rat Leydig cells, which might be influenced by intracellular ROS levels.

关键词： ageing autophagy late onset hypogonadism Leydig cell reactive oxygen species testosterone

Molecular Yin and Yang of erectile function and dysfunction

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Asian Journal of Andrology 2008年第3期10卷 433-440页

作者： Ching-Shwun Lin zhong-cheng xin zhong Wang Guiting Lin Tom E Lue Knuppe Molecular Urology Laboratory Department of Urology School of Medicine University of California San Francisco CA 94143 USA Andrology Center Peking University First Hospital Beijing 100009 China Department of Urology Ninth Hospital Shanghai Second Medical University Shanghai 200011 China

In regard to erectile function, Yin is flaccidity and Yang erection. In the past decade, research has mostly focused on the Yang aspect of erectile function. However, in recent years, the Yin side is attracting increasingly greater attention. This is due to the realization that penile flaccidity is no less important than penile erection and is actively maintained by mechanisms that play critical roles in certain types of erectile dysfunction （ED）; for example, in diabetic patients. In addition, there is evidence that the Yin and Yang signaling pathways interact with each other during the transition from flaccidity to erection, and vice versa. As such, it is important that we view erectile function from not only the Yang but also the Yin side. The purpose of this article is to review recent advances in the understanding of the molecular mechanisms that regulate the Yin and Yang of the penis. Emphasis is given to the Rho kinase signaling pathway that regulates the Yin, and to the cyclic nucleotide signaling pathway that regulates the Yang. Discussion is organized in such a way so as to follow the signaling cascade, that is, beginning with the extracellular signaling molecules （e.g., norepinephrin and nitric oxide） and their receptors, converging onto the intracellular effectors （e.g., Rho kinase and protein kinase G）, branching into secondary effectors, and finishing with contractile molecules and phosphodiesterases. Interactions between the Yin and Yang signaling pathways are discussed as well.

关键词： erectile function erectile dysfunction molecular mechanisms Rho kinase signaling cyclic nucleotide signaling Yin-Yang

Proteomic analysis and miRNA profiling of human testicular endothelial cell-derived exosomes:the potential effects on spermatogenesis

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Asian Journal of Andrology 2022年第5期24卷 478-486页

作者： Wen-Peng Song Sheng-Ji Gu Xiao-Hui Tan Yang-Yang Gu Wei-Dong Song Jian-Yu Zeng zhong-cheng xin Rui-Li Guan Department of Urology Peking University First HospitalBeijing 100034China Institute of Urology Peking UniversityBeijing 100034China Beijing Key Laboratory of Urogenital Diseases(Male)Molecular Diagnosis and Treatment Center Beijing 100034China Department of Dental Implant Center Beijing Stomatological HospitalSchool of StomatologyCapital Medical UniversityBeijing 100050China Department of Radiation Medicine Institute of Systems BiomedicineSchool of Basic Medical SciencesPeking University Health Science CenterBeijing 100191China

Testicular endothelial cells have been found to play an important role in spermatogenesis and fertility,but their mechanism is *** released by various cells are recognized as cell–cell communication mediators during the initiation and progression of many ***,the current study aimed to investigate the protein and miRNA components of human testicular endothelial cell-derived exosomes(HTEC-Exos)and to explore their potential effects on *** this study,HTEC-Exos were first isolated by the ultracentrifugation method,and then identified by nanoparticle tracking analysis,transmission electron microscopy(TEM),and western *** characteristics of HTEC-Exos were examined by liquid chromatography–mass spectrometry and microRNA(miRNA)chip *** analysis was performed to explore the potential role of the exosomal content on spermatogenesis.A total of 945 proteins were identified,11 of which were closely related to spermatogenesis.A total of 2578 miRNAs were *** them,30 miRNAs demonstrated potential associations with male reproductive disorders,such as azoospermia,and spermatogenesis *** particular,11 out of these 30 miRNAs have been proven to be involved in spermatogenesis based on available *** study provides a global view of the proteins and miRNAs from HTEC-Exos,suggesting that HTEC-Exos may function as potential effectors during the process of spermatogenesis.

关键词： exosomes microRNAs proteomics testicular endothelial cells

Further evidence of endogenous hydrogen sulphide as a mediator of relaxation in human and rat bladder

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Asian Journal of Andrology 2013年第5期15卷 692-696页

作者： Jun-Wei Gai Wasilijiang Wahafu Hui Guo Miao Liu Xu-Chang Wang Yun-Xiang Xiao Liang Zhang zhong-cheng xin Jie Jin Department of Urology Peking University First Hospital Beijing 100034 China The Institute of Urology Peking University Beijing 100034 China Department of Urology PLA General Hospital Beijing 100853 China Department of Urology Tsinghua University First Hospital Beijing 100016 China Hedong Center for Disease Control and Prevention Tianjin 300151 China Department of Orthopaedic Surgery Beijing Jishuitan Hospital Fourth Clinical College of Peking University Beijing 100035 China Andrology Center Peking University First Hospital Beijing 100034 China

We investigated the expression of hydrogen sulphide （H2S） in human and rat lower urinary tract （including bladder, prostate and urethra） tissues, and we sought to determine whether H2S induces relaxation of human and Sprague-Dawley （SD） rat bladder strips. Human normal lower urinary tract tissue was obtained for the evaluation of endogenous H2S productivity using a sulphide-sensitive electrode and for the analysis of the expression levels of all three synthases of endogenous H2S, cystathionine β-synthase （CBS）, cystathionine y lyase （CSE） and 3-mercaptopyruvate sulphur transferase （MPST, as known as 3-MST） by Western blot assay. CBS, CSE and MPST were located in human sample slides by immunohistochemistry. Human and male adult SD rat bladder strips were tested for H2S function with a transducer and recorded. All experiments were repeated six times. The endogenous H2S productivity and the H2S synthases had various distributions in the human and rat lower urinary tract tissues and were located in both epithelial and stromal sections. L-cysteine （L-Cys, a substrate of CBS, CSE and MPST） elicited relaxation in a dose-dependent manner on human bladder strips ere-contracted by acetylcholine chloride. This effect could be diminished by the ATe-sensitive potassium ion （KATe） channel blocker glibenclamide （GLB）, the CSE inhibitor DL-propargylglycine （PEG） and the CBS inhibitor hydroxylamine （HA）. H2S and its three synthases were present in the human and rat lower urinary tract tissues and relaxed human and rat bladder strips, which implied that endogenous H2S might play a role in physiological function and pathological disorders of the lower urinary tract symptoms （LUTS） or overactive bladder （OAB）.

关键词： cystathionine β-synthase cystathionine γ lyase detrusor hydrogen sulphide lower urinary tract symptoms 3-mercapto- pyruvate sulphur transferase

Relation of size of seminal vesicles on ultrasound to premature ejaculation

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Asian Journal of Andrology 2017年第5期19卷 554-560页

作者： Zhi-Wei Hong Yu-Ming Feng Yi-Feng Ge Jun Jing Xue-Chun Hu Jia-Ming Shen Long-Ping Peng Bing Yao zhong-cheng xin Center for Reproductive Medicine Jinling Hospital Medical School of Nanjing University Nanjing 210002 China Center for Reproductive Medicine Jinling Hospital Southern Medical University Nanjing 210002 China Andrology Center Peking University First Hospital Peking University Beijing 100009 China

Myriad biological factors have been proposed to explain premature ejaculation （PE）. However, data correlating PE with seminal vesicles （SVs） are sparse. The study aimed to evaluate the relationship between the size of SV and PE. The cross-sectional study included 44 outpatients with PE and 44 volunteers without PE, and the size of SV was compared. Self-estimated intravaginal ejaculatory latency time, the Premature Ejaculation Diagnostic Tool （PEDT）, the International Index of Erectile Function-15, and the National Institutes of Health-Chronic Prostatitis Symptom Index were used for assessment of symptoms. Compared to the control group, the PE group had significantly higher mean anterior-posterior diameter （APD） of SV （P 〈 0.001）. The optimal mean APD of SV cutoff level was 9.25 mm for PE. In the PE group, PEDT was also higher with a mean APD of SV ≥9.25 mm compared with mean APD of SV 〈9.25 mm. PEDT was significantly correlated with the mean APD of SV （r = 0.326, P = 0.031）. The seminal plasma proteins were compared between six PE and six matched control cases by mass spectrometry and it was shown that 102 proteins were at least 1.5-fold up- or down-regulated. Among them, GGT1, LAMC1, and APP were significantly higher in the PE group. These results indicated that men with a larger mean APD of SV might have a higher PEDT score. Transrectal ultrasound of SV should be considered in the evaluation of patients with premature ejaculation. SV might be a potential target for the treatment of patients with PE and ultrasound change in SV.

关键词： mass spectrometry premature ejaculation seminal vesicles size transrectal ultrasound