中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2020, Vol. 15 ›› Issue (1): 120-127.doi: 10.4103/1673-5374.264464

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

脑源性神经营养因子基因修饰脂肪源性干细胞可改善海绵体神经损伤的勃起功能障碍

  


  • 出版日期:2020-01-15 发布日期:2020-05-20
  • 基金资助:

    中国湖北省自然科学基金(2017CFB176)；中国武汉市中心医院基本研究基金(YB16A01)。

Adipose-derived stem cells modified by BDNF gene rescue erectile dysfunction after cavernous nerve injury

Mei Yang1, Jiang-Yang Sun2, Cheng-Cheng Ying3, Yong Wang3, Yong-Lian Guo3   

  1. 1 Department of Endocrinology, Tongren Hospital of Wuhan University (Wuhan Third Hospital), Wuhan, Hubei Province, China
    2 Department of Hepatobiliary Surgery, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province, China
    3 Department of Urology, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province, China
  • Online:2020-01-15 Published:2020-05-20
  • Contact: Cheng-Cheng Ying, MD,dryingcc@126.com.
  • Supported by:
    This study was supported by the Natural Science Foundation of Hubei Province of China, No. 2017CFB176 (to CCY); the Fundamental
    Research Funds for The Central Hospital of Wuhan of China, No. YB16A01 (to CCY).

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摘要：

海绵体神经损伤是前列腺根治性切除术后勃起功能障碍的主要原因。因此前列腺根治性切除术后勃起功能的恢复仍然具有挑战性。课题组以往研究发现，给大鼠海绵体注射脂肪干细胞能够修复受损的海绵体神经，但大鼠的勃起功能不能恢复到正常水平。为此，实验通过慢病毒载体编码大鼠脑源性神经营养因子(BDNF)感染脂肪源性干细胞(ADSCs)来评估其在大鼠海绵体神经损伤模型中的有效性。实验将大鼠随机分为4组：假手术组只分离双侧海绵体神经；双侧海绵体神经损伤(BCNI)组分离双侧海绵体神经并用血管夹钳夹损伤2 min；ADSCGFP 治疗组和ADSCrBDNF 治疗组分别于钳夹损伤2 min后，立即阴茎海绵体内注射感染GFP的大鼠同种异体ADSCs(ADSCGFP)及慢病毒-BDNF感染后的ADSCs(ADSCrBDNF)，注射剂量均为1×106，20 μL。干预后4周，通过海绵体内压来评估勃起功能的恢复情况，然后收集阴茎组织进行组织学和Western Blot检测发现，(1)与BCNI组相比，ADSCGFP 治疗组和ADSCrBDNF 治疗组中大鼠的勃起功能明显恢复，ADSCrBDNF 治疗组提高最明显；(2) ADSCrBDNF 组和ADSCGFP组大鼠背侧神经组织神经型一氧化氮合酶蛋白表达及海绵体中平滑肌与胶原纤维比例明显提高；(3)ADSCrBDNF组的大鼠阴茎组织神经型一氧化氮合酶蛋白表达明显高于ADSCGFP组；(4)上述结果说明，慢病毒-BDNF感染的ADSCs海绵体内注射可有效改善海绵体神经损伤所致勃起功能障碍。实验方案于2017-06-20经武汉大学动物伦理委员会批准，批准号：2017-1638。

orcid: 0000-0002-2738-2809 (Cheng-Cheng Ying)

关键词: 慢病毒, 脂肪干细胞, 勃起功能, 神经再生, 脑源性神经生长因子, 前列腺癌根治术, 海绵体内注射, 神经元型一氧化氮合酶, 海绵体神经损伤, 感染

Abstract: Cavernous nerve injury is the main cause of erectile dysfunction following radical prostatectomy. The recovery of erectile function following radical prostatectomy remains challenging. Our previous studies found that injecting adipose-derived stem cells (ADSCs) into the cavernosa could repair the damaged cavernous nerves, but the erectile function of the treated rats could not be restored to a normal level. We evaluated the efficacy of ADSCs infected with a lentiviral vector encoding rat brain-derived neurotrophic factor (lenti-rBDNF) in a rat model of cavernous nerve injury. The rats were equally and randomly divided into four groups. In the control group, bilateral cavernous nerves were isolated but not injured. In the bilateral cavernous nerve injury group, bilateral cavernous nerves were isolated and injured with a hemostat clamp for 2 minutes. In the ADSCGFP and ADSCrBDNF groups, after injury with a hemostat clamp for 2 minutes, rats were injected with ADSCs infected with lenti-GFP (1 × 106 in 20 μL) and lenti-rBDNF (1 × 106 in 20 μL), respectively. Erectile function was assessed 4 weeks after injury by measuring intracavernosal pressures. Then, penile tissues were collected for histological detection and western blot assay. Results demonstrated that compared with the bilateral cavernous nerve injury group, erectile function was significantly recovered in the ADSCGFP and ADSCrBDNF groups, and to a greater degree in the ADSCrBDNF group. Neuronal nitric oxide synthase content in the dorsal nerves and the ratio of smooth muscle/collagen were significantly higher in the ADSCrBDNF and ADSCGFP groups than in the bilateral cavernous nerve injury group. Neuronal nitric oxide synthase expression was obviously higher in the ADSCrBDNF group than in the ADSCGFP group. These findings confirm that intracavernous injection with ADSCs infected with lenti-rBDNF can effectively improve erectile dysfunction caused by cavernous nerve injury. This study was approved by the Medical Animal Care and Welfare Committee of Wuhan University, China (approval No. 2017-1638) on June 20, 2017.

Key words: adipose-derived stem cells, brain-derived neurotrophic factor, cavernous nerve injury, erectile dysfunction, infection, intracavernous
injection, lentiviral vector, neuronal nitric oxide synthase, radical prostatectomy