人脐带华通胶间充质干细胞对新西兰兔慢性输卵管炎症输卵管结构的修复作用

发布时间：2018-07-11 18:42

本文选题：人脐带间华通胶充质干细胞 + 慢性输卵管炎　；参考：《暨南大学》2015年硕士论文

【摘要】：第一章新西兰兔慢性输卵管炎模型建立及脐带间充质干细胞治疗作用目的:建立新西兰兔慢性输卵管炎症模型,并通过脐带间充质干细胞(WJMSCs)治疗,探讨建模前后及治疗前后输卵管大体及光镜HE染色下的变化。方法:(1)清洁级新西兰雌兔30只,随机分3组,分别为正常对照组6只,模型对照组12只,治疗组12只。模型对照组与治疗组经阴道插管灌注菌液建立炎性损伤模型。(2)建模15天后对模型对照组予无菌生理盐水经两种不同的的给药途径进行对照治疗:分为静脉+阴道灌注组和单纯阴道灌注组,分别标记为模型A组,模型B组,模型A组对新西兰兔进行阴道注射,模型B组对新西兰兔进行生理盐水阴道灌注。(3)建模15天后对治疗组予WJMSCs经两种不同的的给药途径进行治疗:分为全身加局部用药(静脉+阴道灌注)组和局部用药(单纯阴道灌注)组,分别标记为治疗A组,治疗B组。治疗A组对新西兰兔进行阴道注射,治疗B组对新西兰兔进行阴道灌注。(4)所有试验用兔均与最后一次WJMSCs悬液/生理盐水对照治疗后第7天处死。取双侧输卵管间质部,用生理盐水洗涤去除血液,行HE染色。结果:(1)大体观模型对照组慢性炎症反应明显,经WJMSCs治疗后慢性炎症有不同程度减轻。(2)光镜HE染色下观察,模型对照组呈管腔狭窄,粘膜缺失,间质增厚,肌层增厚等典型慢性炎症改变。经WJMSCs治疗后,管腔狭窄缓解,粘膜皱襞结构恢复,间质变薄,肌层稍变薄。结论:(1)通过阴道插管方式向新西兰兔注射大肠埃希菌可成功建立慢性输卵管炎症模型。(2)慢性输卵管炎新西兰兔移植WJMSCs后,从大体及光镜HE染色观察,输卵管的基本结构及周围炎症有不同程度的恢复。第二章WJMSCs修复慢性输卵管炎症损伤的体视学分析目的:利用Masson染色及体视学测定WJMSCs治疗后输卵管粘膜皱襞及肌层变化,探讨相关修复机制。方法:在新西兰兔慢性输卵管炎症模型建立的基础上,通过运用体视学测定单位面积输卵管粘膜皱襞长度及肌层体积密度,比较输卵管慢性炎症情况下及WJMSCs治疗后输卵管粘膜皱襞和肌层结构的变化。结果:(1)模型对照组单位面积输卵管粘膜皱襞长度明显低于正常组,且差异有统计学意义(P=0.000)。经WJMSCs治疗后,单位面积粘膜皱襞长度有不同程度的升高,且差异具有统计学意义(P0.05;P0.05)。其中,治疗B组的粘膜皱襞长度较治疗A组的长(P0.05),但两者的粘膜皱襞长度均小于正常组(P0.05;P0.05)。(2)模型对照组的肌纤维体积密度较正常组增高,差异有统计学意义(P0.05)。治疗B组的肌纤维体积密度较模型对照组有显著性降低(P0.05),治疗A组平均肌纤维体积密度小于模型对照组,但差异无统计学意义(P0.05),治疗B组肌纤维体积密度降低幅度较治疗A组大,差异有统计学意义(P0.05)。但治疗A组与治疗B组均较正常组肌纤维体积密度增大,且差异有统计学意义(P0.05;P0.05)。结论:(1)输卵管慢性炎症可致单位面积输卵管的粘膜皱襞长度减少,经WJMSCs治疗后,单位面积粘膜皱襞长度明显改善,其中单纯局部治疗组治疗效果优于局部全身联合治疗组。(2)输卵管慢性炎症可致输卵管间质部肌纤维体积密度增加,经WJMSCs治疗后肌纤维体积密度减少,而且单纯局部治疗组治疗效果优于局部全身联合治疗组。第三章实时定量聚合酶链式反应测定输卵管相关结构蛋白目的:利用实时定量聚合酶链式反应测定慢性输卵管炎症及WJMSCs治疗后角蛋白及波形蛋白m RNA相关变化,探讨相关修复机制。方法:在新西兰兔慢性输卵管炎症模型建立的基础上,予实时定量聚合酶链式反应测定并比较输卵管慢性炎症情况下及WJMSCs治疗后结构蛋白m RNA表达量的变化。结果:(1)模型对照组的角蛋白m RNA表达量较正常组下降约20倍,差异有统计学意义(P0.05)。治疗A组和治疗B组的角蛋白m RNA表达量较模型对照组均有不同程度的升高(P0.05;P0.05),且治疗B组增高幅度较治疗A组大,差异有统计学意义(P0.05)。但治疗A组与治疗B组均较正常组m RNA表达减少,差异有统计学意义(P0.05;P0.05)。(2)模型对照组的波形蛋白m RNA表达量较正常组增高,差异有统计学意义(P0.05)。治疗A组和治疗B组的波形蛋白m RNA表达量较模型对照组均有不同程度的降低(P0.05;P0.05)。结论:(1)慢性输卵管炎症可致输卵管角蛋白m RNA表达量明显降低,WJMSCs治疗可有效改善角蛋白m RNA的表达。(2)慢性输卵管炎可致输卵管波形蛋白m RNA表达明显增高,WJMSCs治疗可在一定程度上降低波形蛋白m RNA表达。(3)单纯阴道灌注1×106WJMSCs的治疗效果稍优于全身与局部联合用药组。
[Abstract]:Chapter 1 a new model of chronic salpingitis in New Zealand rabbits and the therapeutic effect of umbilical cord mesenchymal stem cells: to establish a new model of chronic tubal inflammation in New Zealand rabbits, and to explore the changes of the oviductal and light microscopy before and after the treatment of the umbilical cord mesenchyme stem cells (WJMSCs). Methods: (1) clean New Zealand. 30 female rabbits were randomly divided into 3 groups, 6 in the normal control group, 12 in the model control group and 12 in the treatment group. The model control group and the treatment group were used to establish the inflammatory injury model by the vaginal intubation liquid. (2) the model control group was given the aseptic saline through two different ways of Administration for 15 days after the modeling, and the control group was divided into venous and vagina. The perfusion group and the simple vaginal perfusion group were labeled as model A group, model B group, model A group were injected with vaginal injection of New Zealand rabbits, model B group was injected with saline and vagina in New Zealand rabbits. (3) after modeling, the treatment group was treated with two different ways of administration to WJMSCs after 15 days: the whole body plus local medication (intravenous plus vaginal irrigation). Group and local medication (simple vaginal perfusion) group were labeled as group A and treated with B group. Group A was treated with vaginal injection of New Zealand rabbits, and group B was treated with vaginal perfusion to New Zealand rabbits. (4) all rabbits were killed at the last seventh days after the last WJMSCs suspension / saline treatment. The blood was washed and removed by HE. Results: (1) the chronic inflammatory reaction in the general model control group was obvious, and the chronic inflammation was reduced in different degrees after WJMSCs treatment. (2) the model control group was observed under HE staining, the model control group showed the typical chronic inflammatory changes, such as the stenosis of the lumen, the deletion of the mucous membrane, the thickening of the interstitium and the thickening of the muscularis. After the treatment of WJMSCs, the tube was treated. Conclusion: (1) a chronic salpingitis model could be successfully established by injecting Escherichia coli into New Zealand rabbits by vaginal intubation. (2) after the transplantation of WJMSCs in New Zealand rabbits with chronic salpingitis, the basic structure and surrounding of the fallopian tubes were observed from the general and light microscope HE staining. The inflammation has different degrees of recovery. Second chapter second the stereological analysis of the repair of chronic oviduct inflammatory injury in chapter second: use Masson staining and stereology to determine the changes of the mucous folds and myometrium of the fallopian tube after the treatment of WJMSCs. Methods: on the basis of the establishment of the chronic oviduct inflammatory model in New Zealand rabbits, the application of this method is to be used. The length of tubal mucosa fold and the volume density of myometrium were measured by stereology, and the changes of the mucosal fold and muscular structure of the fallopian tube were compared in the chronic oviduct and WJMSCs treatment. The results were as follows: (1) the length of the mucous fold of the oviduct in the model control group was significantly lower than that in the normal group, and the difference was statistically significant (P=0.000). After WJMSCs treatment, the length of mucous folds per unit area increased in varying degrees, and the difference was statistically significant (P0.05; P0.05). Among them, the length of the mucosal folds in the treatment group was longer than that of the A group (P0.05), but the length of the mucosa folds was less than that of the normal group (P0.05; P0.05). (2) the volume density of the muscle fiber in the model control group was more than that of the normal group. The volume density of the muscle fiber in the B group was significantly lower than that in the model control group (P0.05). The average muscle fiber volume density in the treatment group was less than that of the model control group (P0.05), but the difference was not statistically significant (P0.05). The reduction of muscle fiber volume density in the group B group was larger than that in the treatment group (P0.), and the difference was statistically significant (P0.). 05) 05). But the volume density of muscle fiber increased in the treatment group and the B group compared with the normal group, and the difference was statistically significant (P0.05; P0.05). Conclusion: (1) the length of the mucous fold of the oviduct can be reduced by chronic oviduct inflammation. After WJMSCs treatment, the length of the mucous membrane folds per unit area is obviously improved, and the therapeutic efficacy of the simple local treatment group is more effective. The results were better than the local combined treatment group. (2) the chronic inflammation of the fallopian tube could cause the increase of the volume density of the interstitial muscle fiber in the fallopian tube, the decrease of the volume density of the muscle fiber after WJMSCs treatment, and the effect of the simple local treatment group was better than the local combined treatment group. The third chapter real-time quantitative polymerase chain reaction was used to determine the tubal related knot. Aim: to determine the related changes in chronic oviduct inflammation and WJMSCs treatment of PVP and vimentin m RNA by real-time quantitative polymerase chain reaction. Methods: Based on the establishment of the chronic oviduct inflammation model in New Zealand rabbits, real-time quantitative polymerase chain reaction was used to determine and compare the fallopian tube slow down. Changes in the expression of structural protein M RNA in the cases of sexual inflammation and WJMSCs treatment. Results: (1) the expression of keratin m RNA in the model control group was about 20 times more than that of the normal group, and the difference was statistically significant (P0.05). The expression of M RNA in the A group and the B group was higher than that in the model group (P0.05; P0.05), and the treatment was also treated. The increase of B group was larger than that of the treatment group A (P0.05), but the expression of M RNA in the treatment group and the B group was less than that in the normal group (P0.05; P0.05). (2) the expression of vimentin m RNA was higher in the model control group than in the normal group, and the difference was statistically significant (P0.05). The expression of RNA was lower than that of the model control group (P0.05; P0.05). Conclusion: (1) chronic oviduct inflammation can significantly reduce the expression of M RNA of tubal protein, WJMSCs treatment can effectively improve the expression of keratin m RNA. (2) chronic salpingitis can increase the expression of oviduct vimentin M RNA expression significantly, WJMSCs treatment can be in one To a certain extent, the expression of vimentin m RNA was reduced. (3) the therapeutic effect of simple vaginal infusion of 1 * 106WJMSCs was slightly better than that of systemic and local combination therapy.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R711.6

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