本文選題：人臍血干細(xì)胞 + 免疫損傷小鼠模型��； 參考：《昆明醫(yī)科大學(xué)》2017年碩士論文

【摘要】：[目的]臨床放化療所致的免疫抑制是患者不良預(yù)后的主要原因,尋找一種修復(fù)免疫抑制安全有效的方法能夠提高患者的治療效果和預(yù)后。本實(shí)驗(yàn)通過建立放療免疫抑制小鼠模型,觀察骨髓干細(xì)胞的輸注對(duì)放療免疫抑制小鼠的修復(fù)作用,再觀察人臍血干細(xì)胞輸注后對(duì)裸鼠的影響。為臍血用于臨床治療放化療所致的造血細(xì)胞損傷和免疫抑制病人提供理論依據(jù)和實(shí)驗(yàn)基礎(chǔ)。[方法]采用放療照射的方法建立小鼠骨髓細(xì)胞損傷和免疫抑制模型,隨后用血細(xì)胞儀檢測(cè)外周血白細(xì)胞、流式細(xì)胞儀檢測(cè)T淋巴細(xì)胞亞群、胸腺細(xì)胞凋亡率,HE染色觀察小鼠骨髓病理切片的變化。將骨髓干細(xì)胞通過尾靜脈輸注至免疫抑制小鼠體內(nèi)后,利用血細(xì)胞分析儀分析外周血血常規(guī),流式細(xì)胞儀檢測(cè)T淋巴細(xì)胞亞群(CD3+、CD4+、CD8+)、ELISA試劑盒檢測(cè)血清免疫球蛋白IgM變化情況,并觀察生存狀態(tài)和存活率。人臍血干細(xì)胞輸注后對(duì)裸鼠的影響:選擇NOD/SCID小鼠,經(jīng)4Gy劑量的x線照射后,隨機(jī)分為實(shí)驗(yàn)組和對(duì)照組,分選臍血有核細(xì)胞,通過尾靜脈輸注至實(shí)驗(yàn)組裸小鼠,流式細(xì)胞儀檢測(cè)人源的CD45、CD3和CD19分子表達(dá),并觀察小鼠存活情況。[結(jié)果]6.0Gy照射組小鼠30d存活率為100%; 8.0Gy照射組小鼠12天后死亡2只,30d存活率為10% ; 10.0Gy照射組小鼠30d存活率為0。因此,確定研究導(dǎo)致小鼠骨髓細(xì)胞和免疫功能損傷的放療照射劑量為8.0Gy。小鼠骨髓干細(xì)胞輸注至放療后小鼠體內(nèi)后,60d存活率為80%,并且移植后小鼠未觀察到移植物抗宿主病(GVHD)等現(xiàn)象,而未經(jīng)輸注干細(xì)胞的小鼠60d存活率為10%;輸注干細(xì)胞后小鼠外周血白細(xì)胞數(shù)、淋巴細(xì)胞比例及CD3+T細(xì)胞亞群較未經(jīng)移植的小鼠高(P0.05);在輸注干細(xì)胞后第15天,CD4+%、CD8+%和免疫球蛋白IgM最低,隨后逐漸升高(P0.05);骨髓干細(xì)胞移植后能降低輻射后小鼠外周血單核細(xì)胞比例及CD4+/CD8+比值(P0.05)。人臍血干細(xì)胞輸注入4Gy劑量的x線照射實(shí)驗(yàn)組NOD/SCID裸小鼠后,同對(duì)照組的裸鼠比較,實(shí)驗(yàn)組裸鼠30d存活率為70%,對(duì)照組裸鼠30d的存活率為10%。并且,通過流式細(xì)胞儀,我們?cè)趯?shí)驗(yàn)組的裸小鼠體內(nèi)檢測(cè)到了人源的CD45、CD3、CD 19分子表達(dá)。[結(jié)論]本實(shí)驗(yàn)成功建立放療致小鼠骨髓細(xì)胞損傷和免疫抑制模型;輸注同種系小鼠骨髓造血干細(xì)胞具有造血重建和免疫功能修復(fù)作用;異種(人)臍血干細(xì)胞能夠顯著延長放射后裸鼠的生存期,在裸鼠實(shí)驗(yàn)研究中提示異種(人)臍血干細(xì)胞有重建造血和免疫系統(tǒng)的征象。
[Abstract]:[objective] Immunosuppression caused by clinical radiotherapy and chemotherapy is the main cause of poor prognosis. Finding a safe and effective method to repair immunosuppression can improve the therapeutic effect and prognosis of patients. In this study, we established an immunosuppressive mouse model of radiotherapy to observe the repair effect of bone marrow stem cell infusion on radiotherapy immunosuppressive mice and the effect of human umbilical cord blood stem cell infusion on nude mice. To provide theoretical and experimental basis for the clinical treatment of hematopoietic cell injury and immunosuppressive patients caused by radiotherapy and chemotherapy. [methods] the model of bone marrow cell injury and immunosuppression in mice was established by radiotherapy. The peripheral blood leukocytes were detected by blood cell analyzer and T lymphocyte subsets were detected by flow cytometry. Thymocyte apoptosis rate and HE staining were used to observe the changes of bone marrow sections in mice. Bone marrow stem cells were infused into immunosuppressive mice via tail vein. The peripheral blood routine was analyzed by hematology analyzer, and the changes of serum immunoglobulin IgM were detected by flow cytometry (FCM) and Elisa kit for detecting T lymphocyte subsets (CD3, CD3, CD 4, CD 4, CD 8). The survival state and survival rate were observed. The effect of human umbilical cord blood stem cell infusion on nude mice: NOD / SCID mice were randomly divided into experimental group and control group after X-ray irradiation of 4 Gy dose. Flow cytometry was used to detect the expression of CD45, CD3 and CD19, and to observe the survival of mice. [results] the 30-day survival rate of mice in 6.0Gy irradiation group was 100, that in 8.0Gy irradiation group was 10% after 12 days, and that in 10.0Gy irradiation group was 0. Therefore, the radiation dose of radiation induced bone marrow cell and immune function injury in mice was determined to be 8.0 GY. The survival rate of mouse bone marrow stem cells was 80% in vivo after radiotherapy, and GV HDD was not observed in mice after transplantation, such as graft-versus-host disease (GV HDD). The survival rate of mice without infusion of stem cells in 60 days was 10 and the number of peripheral blood leukocytes in mice after infusion of stem cells was 10. The percentage of lymphocytes and CD3 T cell subsets were higher than that of untransplanted mice (P 0.05), and on the 15th day after infusion of stem cells, CD4% and IgM were the lowest. After transplantation of bone marrow stem cells, the percentage of peripheral blood monocytes and CD4 / CD8 ratio of mice were decreased gradually, and the ratio of CD _ 4 / CD _ 8 to CD _ 4 / CD _ 8 was decreased after transplantation of bone marrow stem cells. Human umbilical cord blood stem cells were injected with 4 Gy dose of x-ray to irradiated nod / SCID nude mice. Compared with the nude mice of the control group, the survival rate of the nude mice in the experimental group was 70 and that in the control group was 10. By flow cytometry, we detected the expression of human CD45, CD3, and CD19 in nude mice of experimental group. [conclusion] the model of bone marrow cell injury and immunosuppression induced by radiotherapy in mice was successfully established, and the allogeneic bone marrow hematopoietic stem cells were infused with hematopoietic reconstitution and immune function repair. Xenogeneic (human) cord blood stem cells could significantly prolong the survival time of nude mice after radiation. In the experimental study of nude mice, the xenogeneic (human) cord blood stem cells showed signs of reconstitution of hematopoiesis and immune system.
【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R730.5

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 李娟娟;王有為;馬鳳霞;杜文靜;宋寶全;王昕;馮影;田健健;韓忠朝;;比較人臍帶和胎盤間充質(zhì)干細(xì)胞對(duì)小鼠急性移植物抗宿主病的預(yù)防作用[J];中國組織工程研究;2017年05期

2 李曼麗;趙文;高鈺丹;段紅梅;楊朝陽;李曉光;;骨髓間充質(zhì)干細(xì)胞細(xì)胞周期同步化的方法及對(duì)向神經(jīng)細(xì)胞分化的影響[J];中國康復(fù)理論與實(shí)踐;2016年12期

3 陳作觀;刁永鵬;吳志遠(yuǎn);閆盛;李擁軍;;人臍帶血CD34+細(xì)胞誘導(dǎo)小鼠缺血肢體血管生成[J];中國醫(yī)學(xué)科學(xué)院學(xué)報(bào);2016年05期

4 孫自敏;;臍血造血干細(xì)胞移植研究進(jìn)展[J];中國實(shí)用內(nèi)科雜志;2014年02期

5 張玉明;;臍血移植新技術(shù)進(jìn)度[J];南方醫(yī)科大學(xué)學(xué)報(bào);2013年12期

6 李永強(qiáng);劉志輝;胡曉樺;林燕;廖小莉;梁嶸;原春玲;廖思娜;;CD4~+/CD25~+與CD28~+/TCR~+T淋巴細(xì)胞亞群與進(jìn)展期鼻咽癌患者預(yù)后的關(guān)系[J];臨床腫瘤學(xué)雜志;2013年12期

7 孫國軍;趙文靜;陳曦;葉冬霞;李晶;;臍血間充質(zhì)干細(xì)胞的分離培養(yǎng)及生物學(xué)特性[J];中國組織工程研究與臨床康復(fù);2011年14期

8 劉莎;郝文革;黃永蘭;孫新;;臍血總有核細(xì)胞數(shù)對(duì)臍血移植療效的影響[J];中國當(dāng)代兒科雜志;2010年07期

9 張積華;孫康;王妍;田少奇;夏長所;張才龍;于騰波;;臍血間充質(zhì)干細(xì)胞生物學(xué)特性及其成骨成脂分化潛能[J];中國組織工程研究與臨床康復(fù);2010年27期

10 王廣宇;趙芳;朱旅云;;臍血干細(xì)胞的免疫學(xué)特性概述[J];醫(yī)學(xué)綜述;2010年02期

，

本文編號(hào)：2044314