脂肪源成体干细胞移植治疗压力性尿失禁的实验研究  

Experiment study of the therapeutic effects of adipose-derived mesenchymal stem cells for stress incontinence

作　　者：耿鹏[1] 宋红娟[1] 刘亚玲 栾晓梅[1] 黄辉 

机构地区：[1]江苏省徐州市妇幼保健院妇科,徐州221009 [2]江苏省徐州市妇幼保健院病案室,徐州221009

出　　处：《中华细胞与干细胞杂志（电子版）》2015年第3期197-201,共5页Chinese Journal of Cell and Stem Cell（Electronic Edition）

基　　金：江苏省人力资源和社会保障厅六大人才高峰资助项目(ws2011016);江苏省卫生厅资助项目(H201357)

摘　　要：目的通过注射脂肪来源的间充质干细胞至压力性尿失禁(SUI)大鼠,观察大鼠近端尿道尿动力学变化,为其治疗SUI提供实验依据。方法分离、培养雌性SD大鼠脂肪源间充质干细胞,行CD44、CD45、CD90、CD105、CD34免疫荧光检测。细胞经过10μmol/L 5-Aza诱导后采用Western blot方法测定细胞中α-SMA、Desmin、Myosin的表达。以双侧阴部神经阻断的方法,制备大鼠尿失禁模型后将大鼠模型分为3组。Ⅰ组:注射0.2 ml胶原;Ⅱ组:注射0.1 ml胶原和0.1 ml间充质干细胞(1×106/ml)。Ⅲ组为对照组,注射0.2 ml IMDM培养液。2周后大鼠进行最大膀胱容量和漏尿点压力检测。组间差异比较采用t检验。结果流式细胞仪检测显示细胞表达间充质干细胞标志CD44、CD90和CD105,但不表达造血干细胞标志蛋白CD34。细胞在10μmol/L的5-Aza诱导后7 d表达α-SMA、Desmin和Myosin。在大鼠尿失禁模型,胶原组的最大膀胱容量大于对照组[(1.65±0.21)ml vs(1.40±0.12)ml,P=0.035];ADSC组的最大膀胱容量大于胶原组[1.73±0.26)ml vs(1.65±0.21)ml,P=0.042]。胶原组的最大漏尿点压力大于对照组[(41.2±5.2)mm H2O vs(32.2±3.9)mm H2O,P=0.015];ADSC组的最大漏尿点压力大于胶原组[(50.6±6.1)mm H2O vs(41.2±5.2)mm H2O,P<0.05]。结论诱导后的间充质干细胞注射鼠近端尿道后能提高大鼠漏尿点压力和膀胱最大容量,可能成为治疗压力性尿失禁的有效方法。Objective Adipose-derived mesenchymal stem cells were transplanted to rats after stress incontinence was induced and their therapeutic effects were evaluated for possible clinical usage. Methods Mesenchymal stem cells were isolated from adipose tissue of female SD rats. Expression of CD44, CD45, CD90 and CD34 was evaluated using a FACS. Myogenic differentiation was induced with the presence of 10 μmol/L 5-Aza and confirmed by Western blot for α-SMA, Desmin and Myosin. After establishment of model, rats received treatment of either 0.2 ml collagen, 0.1 ml stem cells(1×106/ml)or 0.2 ml IMDM. Leak point pressure and maximal bladder volume were evaluated 2 weeks later. Results Cells were positive for CD44, CD90 and CD105, but negative for CD34. After culture with 10 μmol/L 5-Aza for 7 days,α-SMA, Desmin and Myosin were expressed in cells. Rats in the collagen group had greater bladder volume than rats in the control group(1.65 ± 0.21 ml vs 1.4 ± 0.12 ml, P < 0.05).Rats in the stem cell group had greater bladder volume than rats in the collagen group(1.73 ± 0.26 ml vs 1.65 ± 0.21 ml, P < 0.05). Leak point pressure was greater in the collagen group than in the control group(41.2 ± 5.2 mm H2 O vs 32.2 ± 3.9 mm H2 O, P < 0.05). Leak point pressure was greater in the stem cell group than in the collagen group(50.6 ± 6.1 mm H2 O vs 41.2 ± 5.2 mm H2 O, P < 0.05). Conclusion Adipose-derived mesenchymal stem cells are effective in treating stress urine incontinence after myogenic differentiation.

关 键 词：尿失禁 压力性 间质干细胞 胶原 

分 类 号：R711.59[医药卫生—妇产科学]