本文選題：人臍帶間華通膠充質(zhì)干細(xì)胞 + 慢性輸卵管炎��； 參考：《暨南大學(xué)》2015年碩士論文

【摘要】：第一章新西蘭兔慢性輸卵管炎模型建立及臍帶間充質(zhì)干細(xì)胞治療作用目的:建立新西蘭兔慢性輸卵管炎癥模型,并通過(guò)臍帶間充質(zhì)干細(xì)胞(WJMSCs)治療,探討建模前后及治療前后輸卵管大體及光鏡HE染色下的變化。方法:(1)清潔級(jí)新西蘭雌兔30只,隨機(jī)分3組,分別為正常對(duì)照組6只,模型對(duì)照組12只,治療組12只。模型對(duì)照組與治療組經(jīng)陰道插管灌注菌液建立炎性損傷模型。(2)建模15天后對(duì)模型對(duì)照組予無(wú)菌生理鹽水經(jīng)兩種不同的的給藥途徑進(jìn)行對(duì)照治療:分為靜脈+陰道灌注組和單純陰道灌注組,分別標(biāo)記為模型A組,模型B組,模型A組對(duì)新西蘭兔進(jìn)行陰道注射,模型B組對(duì)新西蘭兔進(jìn)行生理鹽水陰道灌注。(3)建模15天后對(duì)治療組予WJMSCs經(jīng)兩種不同的的給藥途徑進(jìn)行治療:分為全身加局部用藥(靜脈+陰道灌注)組和局部用藥(單純陰道灌注)組,分別標(biāo)記為治療A組,治療B組。治療A組對(duì)新西蘭兔進(jìn)行陰道注射,治療B組對(duì)新西蘭兔進(jìn)行陰道灌注。(4)所有試驗(yàn)用兔均與最后一次WJMSCs懸液/生理鹽水對(duì)照治療后第7天處死。取雙側(cè)輸卵管間質(zhì)部,用生理鹽水洗滌去除血液,行HE染色。結(jié)果:(1)大體觀模型對(duì)照組慢性炎癥反應(yīng)明顯,經(jīng)WJMSCs治療后慢性炎癥有不同程度減輕。(2)光鏡HE染色下觀察,模型對(duì)照組呈管腔狹窄,粘膜缺失,間質(zhì)增厚,肌層增厚等典型慢性炎癥改變。經(jīng)WJMSCs治療后,管腔狹窄緩解,粘膜皺襞結(jié)構(gòu)恢復(fù),間質(zhì)變薄,肌層稍變薄。結(jié)論:(1)通過(guò)陰道插管方式向新西蘭兔注射大腸埃希菌可成功建立慢性輸卵管炎癥模型。(2)慢性輸卵管炎新西蘭兔移植WJMSCs后,從大體及光鏡HE染色觀察,輸卵管的基本結(jié)構(gòu)及周圍炎癥有不同程度的恢復(fù)。第二章WJMSCs修復(fù)慢性輸卵管炎癥損傷的體視學(xué)分析目的:利用Masson染色及體視學(xué)測(cè)定WJMSCs治療后輸卵管粘膜皺襞及肌層變化,探討相關(guān)修復(fù)機(jī)制。方法:在新西蘭兔慢性輸卵管炎癥模型建立的基礎(chǔ)上,通過(guò)運(yùn)用體視學(xué)測(cè)定單位面積輸卵管粘膜皺襞長(zhǎng)度及肌層體積密度,比較輸卵管慢性炎癥情況下及WJMSCs治療后輸卵管粘膜皺襞和肌層結(jié)構(gòu)的變化。結(jié)果:(1)模型對(duì)照組單位面積輸卵管粘膜皺襞長(zhǎng)度明顯低于正常組,且差異有統(tǒng)計(jì)學(xué)意義(P=0.000)。經(jīng)WJMSCs治療后,單位面積粘膜皺襞長(zhǎng)度有不同程度的升高,且差異具有統(tǒng)計(jì)學(xué)意義(P0.05;P0.05)。其中,治療B組的粘膜皺襞長(zhǎng)度較治療A組的長(zhǎng)(P0.05),但兩者的粘膜皺襞長(zhǎng)度均小于正常組(P0.05;P0.05)。(2)模型對(duì)照組的肌纖維體積密度較正常組增高,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。治療B組的肌纖維體積密度較模型對(duì)照組有顯著性降低(P0.05),治療A組平均肌纖維體積密度小于模型對(duì)照組,但差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05),治療B組肌纖維體積密度降低幅度較治療A組大,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。但治療A組與治療B組均較正常組肌纖維體積密度增大,且差異有統(tǒng)計(jì)學(xué)意義(P0.05;P0.05)。結(jié)論:(1)輸卵管慢性炎癥可致單位面積輸卵管的粘膜皺襞長(zhǎng)度減少,經(jīng)WJMSCs治療后,單位面積粘膜皺襞長(zhǎng)度明顯改善,其中單純局部治療組治療效果優(yōu)于局部全身聯(lián)合治療組。(2)輸卵管慢性炎癥可致輸卵管間質(zhì)部肌纖維體積密度增加,經(jīng)WJMSCs治療后肌纖維體積密度減少,而且單純局部治療組治療效果優(yōu)于局部全身聯(lián)合治療組。第三章實(shí)時(shí)定量聚合酶鏈?zhǔn)椒磻?yīng)測(cè)定輸卵管相關(guān)結(jié)構(gòu)蛋白目的:利用實(shí)時(shí)定量聚合酶鏈?zhǔn)椒磻?yīng)測(cè)定慢性輸卵管炎癥及WJMSCs治療后角蛋白及波形蛋白m RNA相關(guān)變化,探討相關(guān)修復(fù)機(jī)制。方法:在新西蘭兔慢性輸卵管炎癥模型建立的基礎(chǔ)上,予實(shí)時(shí)定量聚合酶鏈?zhǔn)椒磻?yīng)測(cè)定并比較輸卵管慢性炎癥情況下及WJMSCs治療后結(jié)構(gòu)蛋白m RNA表達(dá)量的變化。結(jié)果:(1)模型對(duì)照組的角蛋白m RNA表達(dá)量較正常組下降約20倍,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。治療A組和治療B組的角蛋白m RNA表達(dá)量較模型對(duì)照組均有不同程度的升高(P0.05;P0.05),且治療B組增高幅度較治療A組大,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。但治療A組與治療B組均較正常組m RNA表達(dá)減少,差異有統(tǒng)計(jì)學(xué)意義(P0.05;P0.05)。(2)模型對(duì)照組的波形蛋白m RNA表達(dá)量較正常組增高,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。治療A組和治療B組的波形蛋白m RNA表達(dá)量較模型對(duì)照組均有不同程度的降低(P0.05;P0.05)。結(jié)論:(1)慢性輸卵管炎癥可致輸卵管角蛋白m RNA表達(dá)量明顯降低,WJMSCs治療可有效改善角蛋白m RNA的表達(dá)。(2)慢性輸卵管炎可致輸卵管波形蛋白m RNA表達(dá)明顯增高,WJMSCs治療可在一定程度上降低波形蛋白m RNA表達(dá)。(3)單純陰道灌注1×106WJMSCs的治療效果稍優(yōu)于全身與局部聯(lián)合用藥組。
[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.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R711.6

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