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1 ACTA LABORATORIUM ANIMALIS SCIENTIA SINICA August 2016 Vol. 24 No. 4 1, 1, 2, 2, (1., ;2., ) TGF β1, 240 SD, , TGF β1, 20, d 63 d / ELISA TGF 1 TGF β1 21 d, /,TGF β1, ( P > 0 05) TGF β1, TGF β1; ; ; ; Q95 33 A (2016) Doi: / j. issn Comparison of the therapeutic effect of TGF β1 gene therapy and traditional nonsurgical treatment on the rat model of postpartum stress urinary incontinence CHENG Ming jun 1, CAO Yun gui 1,DING Jing xin 2, HUANG Jian 2, (1 Shanghai Jiading Maternal and Child Health Hospital, Shanghai ,China; 2 Obstetrics and Gynecology Hospital of Fudan University, Shanghai ) Abstract Objective To study the effect of TGF β1 gene therapy on the rat model of postpartum stress urinary incontinence and explore a novel non operative treatment of this disease. Methods Two hundred and forty 6 month old SD female rats were used to prepare the model of postpartum stress urinary incontinence by vaginal dilation with a water sac. 148 rats from the 185 successfully prepared model rats were selected, and randomly divided into 5 groups: the TGF β1 gene therapy, clentuterol treatment, electric stimulation therapy, injection of empty vector plasmid, and non treated groups. In addition, 20 normal rats were selected as blank control group. Sneeze test and urodynamic test were conducted, the pelvic floor pubococcygeus muscle contractile force / muscle weight ratio was calculated, serum TGF 1 was detected by ELISA, and TGF 1 protein was detected by immunohistochemistry at 1, 21, 42 and 63 days after the treatment. Results At 21 days after treatment, all the maximum bladder capacity, leak point pressure, and urine or contractile force / muscle weight ratio of the TGF β1 gene therapy group showed even better effects than those of the electrical stimulation group, but the differences were statistically not significant ( P > 0 05). Conclusions TGF β1 gene therapy shows good therapeutic effect on the rat models of postpartum stress urinary incontinence, suggesting that TGF β1 gene therapy may become a new type of non surgical treatment for this disease. Key words TGF β1;gene therapy;stress urinary incontinence, postpartum; Rat Corresponding author: HUANG Jian. E mail: huangjian102@ sina. com [ ] ( :SHDC ); ( :ZK2015B08) [ ] ( ),,,, chengmingjun04@ 163. com [ ] ( ),,, huangjian102@ sina. com

2 Acta Lab Anim Sci Sin,August 2016,Vol. 24 No (postpartum urinary inconti nence,ppui), (stress urinary incontinence, SUI), 10% ~ 60% [1-2],,,SUI,,,, PPUI β1 ( transforming growth factor β1,tgf β1),,,, [3],TGF β1,,,tgf β1, [4],, TGF β1, :SPF,,6 Sprague Dawley (SD) 260, 310 ~ 350 g, [ SCXK ( ) ] [SYXK( ) ] 6,240,20 [5],, 8 Fr 2 cm, 8,, 300 g,8 h :, (AD Instruments BIO Amp CF,QUAD Bridge), POWER LAB Chart v5 2 1, 10 ml / h,, ( ), (10 ml / h) ;, ;,,, SD 128, 32 4 : (TGF β1), ( ),, 20,20 : 1 (TGF β1 ):, TGF β1 DNA 0 1 ml( 50 μg /, ); 0 5 cm 45 o 0 5 cm 2 ( ): ( : 1 5 V, 1 Hz, 0 1 ms, 2 s),,, 20, 3 3 ( ): 30 μg / ml, 3 4 ( ):, 5 ( ):, 0 1 ml /, 1 6 ( ): d 63 d ELISA TGF β1, 0 5 ml, - 80 ; ELISA TGF β1, RB ( Rapidbio),

3 Acta Lab Anim Sci Sin,August 2016,Vol. 24 No. 4 DNM 9602 ; 1 4 TGF β1 : 10 min, 4%, SABC ( ) TGF β1 TGF β1 ( Abcam,ab92486) 1 100; 1 5, ;, ( JH 2), SKY A4, MFLab 3 01 ( ) 2 mm,, 3 6 V 1 Hz 20 ms,, 1 6 SPSS 13 0, Mean ± SD,, P < SD PPUI,, 185, 77 1% ( 185 / 240), 0(0 / 20),, 1 d (P < 0 05 ), ( 1 2), 21 d TGF β1,,, ( P < 0 05 ), ; TGF β1, ( P > 0 05 ) Note. : : 21,P < : Comparison between the clenbuterol, untreatment and null plasmid groups on D21, P < Fig. 1 comparison of the maximum bladder capacity of the rats 2 2 TGF β1 TGF β1 ( 3),TGF β1, 1 d TGF β1 (P < 0 05),, TGF β1 TGF β1, (P < 0 05) 2 3 TGF βl, TGF βl,, ;,TGF βl 1 d TGF βl TGF βl, TGF β1 ( 4)

4 中国实验动物学报 ２０１６ 年 ８ 月第 ２４ 卷第 ４ 期 注 Ａｃｔａ Ｌａｂ Ａｎｉｍ Ｓｃｉ Ｓｉｎ Ａｕｇｕｓｔ ２０１６ Ｖｏｌ ２４ Ｎｏ ４ 与第 ２１ 天同次检测的药物治疗组 未治疗组和空载质粒组比较 Ｐ ０ ０５ 图２ Ｎｏｔｅ Ｆｉｇ ２ 注 Ｃｏｍｐａｒｉｓｏｎ ｏｆ ｔｈｅ ｌｅａｋ ｐｏｉｎｔ ｐｒｅｓｓｕｒｅ ｏｆ ｔｈｅ ｒａｔｓ 与第 １ 天同次检测的药物治疗组 电刺激组 未治疗组和空载质粒组比较 Ｐ ０ ０５ 图３ Ｎｏｔｅ 大鼠的漏尿点压力 Ｃｏｍｐａｒｉｓｏｎ ｂｅｔｗｅｅｎ ｔｈｅ ｃｌｅｎｂｕｔｅｒｏｌ ｕｎｔｒｅａｔｍｅｎｔ ａｎｄ ｎｕｌｌ ｐｌａｓｍｉｄ ｇｒｏｕｐｓ ｏｎ ｄａｙ２１ Ｐ ０ ０５ 血清 ＴＧＦ β１ 含量 Ｃｏｍｐａｒｉｓｏｎ ｂｅｔｗｅｅｎ ｔｈｅ ｃｌｅｎｂｕｔｅｒｏｌ ｐｈｙｓｉｃａｌ ｅｌｅｃｔｒｉｃ ｓｔｉｍｕｌａｔｉｏｎ ｕｎｔｒｅａｔｍｅｎｔ ａｎｄ ｎｕｌｌ ｐｌａｓｍｉｄ ｇｒｏｕｐｓ ｏｎ ｄａｙ２１ Ｐ ０ ０５ Ｆｉｇ ３ 图４ Ｆｉｇ ４ Ｃｏｍｐａｒｉｓｏｎ ｏｆ ｔｈｅ ｓｅｒｕｍ ＴＧＦ β１ ｌｅｖｅｌｓ ｏｆ ｔｈｅ ｇｒｏｕｐｓ 注 Ａ ＴＧＦ β１ 组 Ｂ 未治疗组 Ｃ 对照组 耻尾肌组织 ＴＧＦ β１ 表达 均为治疗后 １ ｄ ２００ Ｎｏｔｅ Ａ ＴＧＦ β１ ＴＧＦ β１ ｇｒｏｕｐ Ｂ Ｕｎｔｒｅａｔｍｅｎｔ ｇｒｏｕｐ Ｃ Ｃｏｎｔｒｏｌ ｇｒｏｕｐ ２００ Ｅｘｐｒｅｓｓｉｏｎ ｏｆ ＴＧＦ β１ ｉｎ ｔｈｅ ｐｕｂｏｃｏｃｃｙｇａｌ ｍｕｓｃｌｅ ｔｉｓｓｕｅ ａｔ ｏｎｅ ｄａｙ ａｆｔｅｒ ｔｒｅａｔｍｅｎｔ ４１１

5 Acta Lab Anim Sci Sin,August 2016,Vol. 24 No / / ( 5),, 1 d ( P > 0 05), 21 d TGF β1, (P < 0 05 ); TGF β1, (P > 0 05 ) Note. : 21,P < : Comparison between the clenbuterol, untreatment and null plasmid groups on day21, P < Fig. 5 Ratios of muscle contractile force / weight in the rat groups 3,, ( kegel ) [6,7],, [8],, [9],,,, [10] 2008 Carr PPUI, PPUI, [11-13],, ; SUI, [14,15] ( transforming growth factor,tgf), [3] ;,TGF β ECM, TGF βl [16] PPUI, ECM, PFD [17],TGF β1, [18], TGF β1 PFD TGF β1 PPUI,, TGF β1 PPUI, 1 d,, TGF β1,,,

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