Castiglione Fabio, Hedlund Petter, Weyne Emanuel, Hakim Lukman, Montorsi Francesco, Bivalacqua Trinity J, De Ridder Dirk, Milenkovic Uros, Ralph David, Garaffa Giulio, Muneer Asif, Joniau Steven, Albersen Maarten
Laboratory for Experimental Urology, Organ Systems, Department of Development and Regeneration, University of Leuven, Leuven, Belgium; The Institute of Urology, University College of London Hospital, London, UK; Division of Oncology/Unit of Urology, Urological Research Institute, IRCCS Ospedale San Raffaele, Milan, Italy.
Department of Clinical and Experimental Pharmacology, Lund University, Sweden; Division of Drug Research, Department of Medical and Health Sciences, Linköping University, Sweden.
Sex Med. 2019 Mar;7(1):94-103. doi: 10.1016/j.esxm.2018.09.003. Epub 2018 Nov 30.
Previous studies have shown that the injection of adipose tissue-derived stem cells (ADSCs) into the tunica albuginea (TA) during the active phase of Peyronie's disease (PD) prevents the development of fibrosis.
To investigate, using an animal model, whether local injection of human ADSCs (hADSCs) can alter the degree of fibrosis in the chronic phase of PD.
27 male, 12-week-old rats were divided into 3 equal groups: sham, PD without treatment, and PD treated with hADSCs 1 month after disease induction. Sham rats underwent 2 injections of vehicle into the TA 1 month apart. PD rats underwent transforming growth factor β1 (TGFβ1) injection and injection of vehicle 1 month later. PD-hADSC rats underwent TGFβ1 injection followed by 1 million hADSCs 1 month later. 1 week after treatment, n = 3 animals/group were euthanized, and the penises were harvested for quantitative polymerase chain reaction. 1 month after treatment, the other animals, n = 6 per group, underwent measurement of intracavernous pressure (ICP) and mean arterial pressure (MAP) during electrostimulation of the cavernous nerve. After euthanasia, penises were again harvested for histology and Western blot.
The primary outcome measures included (a) gene expression at one week post-injection; (b) measurement of ICP/MAP upon cavernous nerve stimulation as a measure of erectile function; (c) elastin, collagen I and III protein expression; and (d) Histomorphometric analysis of the penis. Means where compared by analysis of variance (ANOVA) followed by a Student-Newman-Keuls test for post hoc comparisons or Mann-Whitney test when applicable.
No significant difference was noted in ICP or ICP/MAP in response to cavernous nerve electrostimulation between the 3 groups at 2.5, 5, and 7.5 V (P > .05 for all voltages). PD animals developed tunical and subtunical areas of fibrosis with a significant upregulation of collagen III protein. The collagen III/I ratio was higher in the PD (4.6 ± 0.92) group compared with sham (0.66 ± 0.18) and PD-hADSC (0.86 ± 0.06) groups (P < .05) These fibrotic changes were prevented when treated with hADSCs. Compared with PD rats, PD-hADSC rats demonstrated a decreased expression of several fibrosis-related genes.
Injection of hADSCs reduces collagen III expression in a rat model of chronic PD. Castiglione F, Hedlund P, Weyne E, et al. Intratunical Injection of Human Adipose Tissue-Derived Stem Cells Restores Collagen III/I Ratio in a Rat Model of Chronic Peyronie's Disease. Sex Med 2019;7:94-103.
先前的研究表明，在佩罗尼氏病（PD）的活动期将脂肪组织来源的干细胞（ADSCs）注入白膜（TA）可预防纤维化的发展。
使用动物模型研究局部注射人ADSCs（hADSCs）是否能改变PD慢性期的纤维化程度。
将27只12周龄雄性大鼠平均分为3组：假手术组、未治疗的PD组和疾病诱导1个月后用hADSCs治疗的PD组。假手术组大鼠在1个月内分两次向TA注射赋形剂。PD组大鼠先注射转化生长因子β1（TGFβ1），1个月后注射赋形剂。PD-hADSC组大鼠先注射TGFβ1，1个月后注射100万个人ADSCs。治疗1周后，每组处死3只动物，摘取阴茎进行定量聚合酶链反应。治疗1个月后，每组另外6只动物在海绵体神经电刺激期间测量海绵体内压（ICP）和平均动脉压（MAP）。安乐死后，再次摘取阴茎进行组织学检查和蛋白质印迹分析。
主要观察指标包括：（a）注射后1周的基因表达；（b）海绵体神经刺激时ICP/MAP的测量，作为勃起功能的指标；（c）弹性蛋白、I型和III型胶原蛋白的蛋白表达；（d）阴茎的组织形态计量分析。采用方差分析（ANOVA）进行均值比较，随后进行Student-Newman-Keuls检验进行事后比较，或在适用时进行Mann-Whitney检验。
在2.5、5和7.5V时，3组之间在海绵体神经电刺激时的ICP或ICP/MAP方面未观察到显著差异（所有电压下P>.05）。PD组动物出现白膜和白膜下纤维化区域，III型胶原蛋白蛋白显著上调。与假手术组（0.66±0.18）和PD-hADSC组（0.86±0.06）相比，PD组（4.6±0.92）的III型胶原蛋白/I型胶原蛋白比值更高（P<.05）。用hADSCs治疗可预防这些纤维化改变。与PD大鼠相比，PD-hADSC大鼠的几种纤维化相关基因表达降低。
在慢性PD大鼠模型中，注射hADSCs可降低III型胶原蛋白的表达。Castiglione F，Hedlund P，Weyne E等。在慢性佩罗尼氏病大鼠模型中，白膜内注射人脂肪组织来源的干细胞可恢复III型胶原蛋白/I型胶原蛋白比值。性医学2019；7：94-103。
