脂肪干细胞联合体外冲击波治疗大鼠勃起功能障碍疗效研究

发布时间：2018-04-23 09:38

本文选题：脂肪干细胞 + 低能量体外冲击波　；参考：《郑州大学》2017年硕士论文

【摘要】：研究背景勃起功能障碍(erectile dysfunction,ED)严重影响生活质量,药物疗法对一些难治性ED治疗效果欠佳,也无法改变ED的病理状态,只能减缓症状。脂肪干细胞(adipose tissue-derived stem cell,ADSC)和低能量体外冲击波疗法(low-intensity shock wave therapy,LI-ESWT)是治疗ED的新方法。近十年来,这两种治疗方法得到了巨大发展。在亚临床研究中,这两种方法对ED也都取得了不错的治疗效果,但是无论哪一种方法都无法治愈ED,也无法使勃起功能达到正常水平。亚临床研究显示这两种方法对于ED的治疗机制有着相似的地方,都能改善ED病理状态。考虑到二者的共同之处,本研究从这两种方法联合的角度探讨二者对于勃起功能障碍的治疗效果。目的评价ADSC联合LI-ESWT治疗ED的疗效,探索ADSC和LI-ESWT之间的交互作用,阐明ADSC联合LI-ESWT治疗ED的机制。方法取4周龄无特定病原菌(specific pathogen free,SPF)级SD(Sprague Dawley)雄性大鼠腹股沟处脂肪,用于培养ADSC。噻唑蓝(MTT)法测定ADSC生长曲线,成脂、成骨分化用于鉴定ADSC。150只8周龄SPF级SD雄性大鼠随机分成正常对照组和交互作用分析组,正常组未给予任何处理;交互作用分析组腹腔注射链脲佐菌素(streptozotocin,STZ)构建Ⅰ型糖尿病模型,糖尿病大鼠进行阿扑吗啡(apomorphine,APO)实验,APO实验阴性大鼠随机分成ED模型组,给予海绵体移植磷酸盐缓冲液(phosphate buffer saline,PBS);ADSC治疗组,给予海绵体移植ADSCs悬液;LI-ESWT治疗组,给予海绵体冲击波治疗;ADSC联合LI-ESWT治疗组,在移植ADSCs1天后给予海绵体冲击波治疗。干预4周后,测量每组大鼠海绵体内压(intracavernous pressure,ICP)及平均动脉压(mean arterial pressure,MAP)评估治疗效果,分析LI-ESWT和ADSC间交互作用。取大鼠阴茎中段进行免疫组织化学(immunohistochemistry,IH)实验来探索可能的治疗机制。结果1、ADSC增殖能力强,能诱导分化为脂肪细胞和成骨细胞,3代ADSC的倍增时间为26.26±3.12h。2、最终共98只大鼠完成实验,正常组19只,ED模型组19只,ADSC治疗组20只,LI-ESWT治疗组19只,联合治疗组21只。在10V刺激下五组间ICP/MAP值分别为0.69±0.096,0.25±0.096,0.44±0.13,0.42±0.14和0.55±0.082,五组间ICP/MAP差异有统计学意义(p0.05);ADSC组、LI-ESWT组和联合治疗组ICP/MAP明显高于ED模型组;联合治疗组ICP/MAP值高于ADSC组和LI-ESWT组。LI-ESWT和ADSC间交互作用无统计学意义(P=0.212),大鼠的血糖水平和ICP/MAP的spearman相关系数为-0.316,P0.05,二者呈负相关。3、ADSC治疗组、LI-ESWT组和联合治疗组?-SMA,nNOS和vWF的表达均高于ED模型组,P0.05。结论ADSC和LI-ESWT疗法能够改善大鼠勃起功能,二者联合对大鼠勃起功能较单独使用时效果更好,二者之间不存在交互作用。ADSC和LI-ESWT均可以提高ED大鼠阴茎海绵体?-SMA,n NOS和v WF的表达。
[Abstract]:Background erectile dysfunctional EDV (erectile dysfunctional EDV) has a serious impact on the quality of life. The therapeutic effect of drug therapy on refractory Ed is poor, and it can not change the pathological state of Ed, but can only alleviate the symptoms. Adipose tissue-derived stem (ADSCC) and low-intensity shock wave therapy (LI-ESWTT) are new methods for the treatment of Ed. In the last ten years, these two treatments have been greatly developed. In subclinical studies, both methods have achieved good results in the treatment of Ed, but neither method can cure EDand nor achieve normal erectile function. Subclinical studies have shown that these two methods are similar to each other in the treatment mechanism of Ed and can improve the pathological state of Ed. In view of their common features, this study explored the therapeutic effects of the two methods on erectile dysfunction from the point of view of the combination of the two methods. Objective to evaluate the efficacy of ADSC combined with LI-ESWT in the treatment of Ed, explore the interaction between ADSC and LI-ESWT, and elucidate the mechanism of ADSC combined with LI-ESWT in the treatment of Ed. Methods the inguinal fat of 4-week-old SD(Sprague Dawley male rats without specific pathogen was used to culture ADSCs. The growth curve of ADSC was determined by thiazolyl blue method, and the adipogenic and osteogenic differentiation was used to identify the 8-week-old SPF male rats of ADSC.150. They were randomly divided into normal control group and interactive analysis group, and the normal group was not given any treatment. The model of type I diabetes was established by intraperitoneal injection of streptozotocin (STZ) in the interaction analysis group. The rats with apomorphine apomorphine (APO) were randomly divided into Ed model group and treated group with phosphate buffer solution phosphate buffer for cavernosum transplantation. The cavernous body transplantation ADSCs suspension LI-ESWT treatment group, cavernous shock wave therapy combined with LI-ESWT treatment group, after ADSCs1 day, the cavernous body shock wave treatment group. After 4 weeks of intervention, intracavernous pressure (ICP) and mean arterial pressure (mean arterial pressure MAPP) were measured in each group, and the interaction between LI-ESWT and ADSC was analyzed. The middle part of rat penis was used to study the possible therapeutic mechanism by immunohistochemistry IH. Results 1the multiplication time of ADSC was 26.26 卤3.12h.2.The multiplication time of ADSC was 26.26 卤3.12h.2. finally, the experiment was completed in 98 rats, 19 rats in normal group and 19 rats in ADSC-treated group (n = 19) treated with LI-ESWT, 19 rats were treated with LI-ESWT, 19 rats in normal group were treated with LI-ESWT, 19 rats in normal group were treated with LI-ESWT. There were 21 rats in the combined treatment group. Under 10 V stimulation, the ICP/MAP values of the five groups were 0.69 卤0.0966 0.25 卤0.096 卤0.44 卤0.130.42 卤0.14 and 0.55 卤0.082, respectively. The ICP/MAP of the LI-ESWT group and the combined treatment group were significantly higher than that of the Ed model group. The ICP/MAP value of combined treatment group was higher than that of ADSC group and LI-ESWT group. There was no significant difference in the interaction between LI-ESWT and ADSC. The correlation coefficient between blood glucose level and spearman of ICP/MAP in rats was -0.316, P 0.05. The expression of SMAnNOS and vWF in LI-ESWT group and combined treatment group was negatively correlated with that in ADSC group. All of them were higher than that of Ed model group (P 0.05). Conclusion the combination of ADSC and LI-ESWT can improve erectile function in rats, and the combination of them can improve the expression of NOS and vWF in cavernosum cavernosum of Ed rats. There is no interaction between them.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R698

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