| Objectives: The SUI rat model was established to detect the change of rats’urodynamic and observe the histology alteration of urethra and tissues around it.Immunohistochemical method and image analysis were used to analyze theexpression of the three factors, i.e. desmin, myosin heavy chain and PGP9.5, inurethra. Meanwhile the effect on mesenchymal stem cells treating SUI were alsostudied through these two methods. It finally discussed two different ways of stemcells transplantation to guide clinical application。Materials and methods:1．SD female rat graded as clean were selected and Ratswere induced to develop SUI by postpartum vaginal balloon dilation and bilateralovariectomy，Then the models was validated through the measurement of the leakPoint Pressure (LPP)、maximum bladder capacity of experimental animals (MBC)and the Positive rate of sneeze testing and histologic sections,the Urinary sphinctermicrodissection of rat was researched by HE staining Masson staining and Malloryspecial staining.2．After modeling successfully, the twenty rats were randomized into three sets:intravenous injection (group A, six rats), urethral injections (group B, eight rats),reference group (group C, six rats) and normal group (group D, six rats). MSCs weretransplanted into SUI rat at5*106cells per one rat by tail intravenous tubes andurethra part injection methods. After for weeks and six weeks, the urodynamicindexes and positive rate of sneezing experiment were detected separately by meansof BL-410biological mechanism experiment system to evaluation the urodynamicvariation.3．All rats were sacrificed after six weeks of injection to get experiment materials ofurethras and tissue around it. Through histomorphometric property and image analysis,It could observe the change of urethra and tissue around it, and evaluate contents ofdesmin、MHC and pgp9.5in order to quantitatively calculate the positive area ratio ofstriated muscle to the whole urethra and sphincter urethra to the entire urethra.Thenwe can Further understand the efficacy and possible mechanisms for HUMSCs therapy for SUI。Results：1.SD female urethral sphincter was unevenly distributed in the entire urethra,In the middle part of the urethral,the rhabdosphincter become be more circular andthick。The SUI model of simulating human pregnancy, dystocia injury and low levelestrogen after castration could be established. From the morphology observation ofurethra and tissue around it, it could obtain: muscular layer of sphincter urethra beingdecreased, loose and disorder arrangement, part of muscle having crack andvacuolation, utrophin desmin and MHC staining reduction even being missed.2.LPP is the most important indicator of the reaction urinary function， Itdemonstrates that the urinary control function is recovered to some extent when LPProse.After four weeks LPP was significantly increased By HUMSCs-treated group，LPP were18.15±1.21mmHg，18.79±1.48mmHg,13.55±1.43mmHg，22.83±2.17mmHg in group A，group B，group C and group D respectively。And two weekslater the LPP changed into18.86±1.14mmHg,19.40±0.85mmHg，11.82±0.91mmHg,24.36±3.26mmHg for the four different groups, respectively. There isstatistic difference for pairwise comparisons (P<0.01), and C group had theminimum LPP content. Pairwise comparisons among A, B, and D groups hadstatistic significance, but no sense between A and B group（P>0.05）. LPP among allgroups did not have any statistic significance after HUMSCs treatment neither withinfour weeks nor six weeks （P>0.05）.3.urethral sphincter layer was apparently strengthened and the function was alsosignificantly improved after HUMSCs treatment. Image analysis indicated thatpositive area of urethral striated muscle and sphincter was augmented distinctly, thenumber of PGP9.5was increased remarkably compared with positive reference group.The positive area ratio of Urethral striated muscle of A, B, C and D groups was(15.3±3.77)%,(15.2±4.79)%,(16.49±2.4)%,(9.55±2.06)%; The positive area ratio ofUrethral sphincter was (35.14±5.38)%,(36.88±3.93)%,(22.46±3.19)%,(37.9±1.80)%;and the number of PGP was7.75±1.26，8.14±1.68，5.2+2.05，10.25+1.89respectivelyfor four groups aforementioned. The pairwise comparision among the test groups for the three factors all had statistic deviation（P<0.01），and the minimum positive areaof Urethral striated muscle came from C group. There was statistic significance eithercompared with the other three groups（P<0.01），however no statistic meaning existedin the pairwise comparison among A, B and D groups.Conclusion:1.In the study，SUI animal model was successfully established throughpregnancy, delivery vaginal expansion and bilateral spayed castration, paving the wayto further research.2. SUI appeared due to the sphincter muscle damage and nerve fiber reduction, whichhinted the change of urethral structure and function as the direct factor of inducingSUI.3. The content of utrophindesmin, MHC and the number of axons markers PGP9.5were improved by means of transplanting cells to SUI model.4. The two kinds of treatment i.e. local injection treatment and intravenous injectiontreatment had few differences.5. HUMSCs transplanted to SUI rats reproduce urethral sphincter tissue or secretedrelative factors to facilitate the regeneration of damaged muscle and nerve in order toshrinking function of urethra, recover weak urethra sphincter to realize thepurpose of improving urethra function. |
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