本文選題：骨髓間充質(zhì)干細(xì)胞　切入點(diǎn)：60Coγ射線(xiàn)　出處：《鄭州大學(xué)》2014年碩士論文

【摘要】：研究背景及目的： 不育是泌尿男科門(mén)診常見(jiàn)疾病，影響約15%的夫婦，并且男性因素作為主要因素的約占30%，而大約40%的夫婦雙方都有問(wèn)題。雄性不育的發(fā)病機(jī)制尚未完全明確，病因明確者采用對(duì)癥治療，而原因不明者，多采用經(jīng)驗(yàn)治療，效果多不滿(mǎn)意。近些年來(lái)，輔助生育技術(shù)發(fā)展迅速，為廣大不育患者提供了福音，但輔助生育技術(shù)違反了生物界自然選擇的規(guī)律，而且輔助生育技術(shù)都是有創(chuàng)的，經(jīng)濟(jì)負(fù)擔(dān)較大，并具有潛在的健康風(fēng)險(xiǎn)。而且對(duì)于那些一方不能產(chǎn)生有功能配子的夫婦，除了求助于異體供體沒(méi)有其他的治療方案。 近些年來(lái)隨著干細(xì)胞在雄性不育中的研究深入，發(fā)現(xiàn)干細(xì)胞可以分化為生殖細(xì)胞，為雄性不育的治療和發(fā)病機(jī)制研究提供了新的途徑。本文就骨髓間充質(zhì)干細(xì)胞移植是否對(duì)大鼠生精障礙研究具有治療作用，并初步探討其潛在的機(jī)制，為臨床上雄性不育的治療提供更多的治療方法。 方法： 75只健康雄性Wistar大鼠隨機(jī)均分為5組，每組15只。A組為正常對(duì)照組,B組為低劑量照射對(duì)照組，C組高劑量照射對(duì)照組，D組為低劑量照射治療組，E組為高劑量照射治療組。B組和D組兩組大鼠均以60Co γ射線(xiàn)局部照射1次，C組和E組大鼠均局部照射2次（間隔一周照射）。造模成功后，D組和E組給予大鼠睪丸骨髓間充質(zhì)干細(xì)胞移植，飼養(yǎng)8周。8周后，稱(chēng)取體重及睪丸重量，并計(jì)算睪丸指數(shù)。采用ELISA法檢測(cè)血清中FSH、LH、T的水平，應(yīng)用HE染色觀(guān)察大鼠睪丸組織形態(tài)學(xué)變化，應(yīng)用RT-PCR檢測(cè)GFP、Inhibin B、P450scc和VASA表達(dá)情況,雙重免疫熒光檢測(cè)GFP和VASA的表達(dá)情況。 結(jié)果： 照射對(duì)照組體重及睪丸指數(shù)均明顯下降，并具有統(tǒng)計(jì)學(xué)意義（P＜0.05），而治療組體重則與正常對(duì)照組相比無(wú)統(tǒng)計(jì)學(xué)意義（P＞0.05），而睪丸指數(shù)仍明顯下降，并具有統(tǒng)計(jì)學(xué)意義（P＜0.05）。性激素方面，模型組及治療組血清FSH顯著提高，T含量明顯降低，LH升高，差異均有統(tǒng)計(jì)學(xué)意義（P＜0.05）；治療后，F(xiàn)SH、LH下降，T升高，兩組LH和T變化均具有統(tǒng)計(jì)學(xué)意義（P＜0.05）；而高劑量治療組FSH下降無(wú)統(tǒng)計(jì)學(xué)意義（P＞0.05）。睪丸組織病理學(xué)方面，照射對(duì)照組曲細(xì)精管結(jié)構(gòu)破壞明顯，可見(jiàn)極少量的支持細(xì)胞和精原細(xì)胞，以高劑量對(duì)照組破壞更為明顯；治療后，低劑量干細(xì)胞組大鼠生精功能有所恢復(fù)，而高劑量干細(xì)胞組生精功能恢復(fù)欠佳。RT-PCR示:照射模型組睪丸組織中InhibinB、P450scc和VASA mRNA幾乎不表達(dá)；給予治療后，低劑量治療組大鼠InhibinB、P450scc和VASA mRNA表達(dá)水平升高，而高劑量治療組僅有P450scc表達(dá)水平升高，而VASA和Inhibin B表達(dá)水平改變不明顯。雙重免疫熒光檢測(cè)VASA和GFP結(jié)果同RT-PCR。 結(jié)論： 骨髓間充質(zhì)干細(xì)胞移植后可以明顯改善生精功能輕度受損大鼠睪丸的生精功能，而對(duì)于損傷較重，支持細(xì)胞恢復(fù)欠佳者生精功能恢復(fù)不明顯。
[Abstract]:Background and objectives:. Infertility is a common disease in urological andrology clinics, affecting about 15% of couples, and male factors account for about 30% of the main factors, and about 40% of couples have problems with each other. The pathogenesis of male sterility is not completely clear. Those with definite etiology are treated with symptomatic treatment, while those with unknown causes are treated with experience, and the results are not satisfactory. In recent years, assisted reproductive technology has developed rapidly, providing good news for the vast number of infertile patients. But assisted reproductive technology violates the laws of natural selection in the biological world, and it is invasive, financially burdensome, and has potential health risks. And for couples who are not able to produce functional gametes on one side, There is no treatment other than the use of allogeneic donors. In recent years, with the further study of stem cells in male sterility, it has been found that stem cells can differentiate into germ cells. This study provides a new approach for the treatment of male sterility and the study of pathogenesis of male sterility. In this paper, we discuss the therapeutic effect of bone marrow mesenchymal stem cell transplantation on rat spermatogenesis disorder, and explore its potential mechanism. To provide more clinical treatment for male sterility. Methods:. 75 healthy male Wistar rats were randomly divided into 5 groups. There are 15 rats in each group. Group A is normal control group, group B is low dose irradiation, control group, group C, group C, high dose irradiation control group, group D, low dose radiation treatment group, group E, high dose irradiation treatment group, group B and group D, all rats are treated with 60Co 緯 rays. Rats in group C and group E were given local irradiation twice (interval of one week). Group D and group E were transplanted with mesenchymal stem cells (MSCs) of testis after successful modeling. After 8 weeks of feeding, weight of testis and weight of testis were measured and testicular index was calculated. ELISA method was used to detect the level of FSHLHX T in serum, HE staining was used to observe the morphological changes of testis, and RT-PCR was used to detect the expression of P450 SCC and VASA in rat testis. Double immunofluorescence was used to detect the expression of GFP and VASA. Results:. The body weight and testicular index of the irradiated control group were significantly decreased (P < 0.05), while the weight of the treatment group was not significantly higher than that of the normal control group (P > 0.05), while the testicular index was still significantly decreased (P < 0.05). In model group and treatment group, serum FSH level was significantly increased and the level of LH was significantly decreased (P < 0.05), and the serum FSH level was significantly lower than that in control group (P < 0.05). The changes of LH and T in both groups were statistically significant (P < 0.05), but the decrease of FSH in high-dose treatment group was not statistically significant (P > 0.05). In testicular histopathology, the structure of seminiferous tubules was destroyed obviously in the irradiated control group, and a very small number of Sertoli cells and spermatogonocytes were observed. After treatment, the spermatogenic function of rats in the low-dose stem cell group was recovered, while the spermatogenic function in the high-dose stem cell group was poor. RT-PCR showed that there was almost no expression of VASA mRNA and Inhibin Bon P450scc in the testis of the model group. After treatment, the expression levels of Inhibin Bon P450 SCC and VASA mRNA increased in the low dose group, but only in the high dose group, but the expression level of VASA and Inhibin B did not change significantly. The results of double immunofluorescence detection of VASA and GFP were similar to those of RT-PCR. Conclusion:. After transplantation of bone marrow mesenchymal stem cells, the spermatogenic function of the testis of the rats with slightly impaired spermatogenic function was obviously improved, but the spermatogenic function of the rats with severe injury and poor recovery of Sertoli cells was not obvious.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R698.2

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