發(fā)布時(shí)間：2018-04-16 22:37

  本文選題：外泌體 + MDSC　； 參考：《重慶醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:研究小鼠腎癌生長被干擾素α2b抑制及其潛在的作用機(jī)制。方法:1.建立腎癌移植瘤模型,將腎癌移植瘤模型分為對(duì)照組和干擾素α2b組,分別給予空白處理和干擾素α2b干預(yù)處理。建模后兩組荷瘤小鼠移植瘤體積與重量進(jìn)行觀察監(jiān)測,切取移植瘤作HE染色。2.外泌體通過超濾蔗糖密度梯度離心法提取,其形態(tài)及數(shù)量的變化通過透射電鏡鑒定。3.在干擾素α2b組與對(duì)照組中,淋巴細(xì)胞應(yīng)用FCM檢測其所占百分比。4.在干擾素α2b組與對(duì)照組中,髓源性抑制性細(xì)胞應(yīng)用FCM檢測其所占百分比。5.BALB/c小鼠實(shí)驗(yàn)?zāi)Ｐ统晒?chuàng)建,將其分為生理鹽水組、外泌體組和Renca細(xì)胞組,其后每隔10天荷瘤小鼠骨髓中骨髓源性抑制性細(xì)胞比例應(yīng)用FCM檢測。結(jié)果:1.腎癌移植瘤模型與BALB/c小鼠實(shí)驗(yàn)?zāi)Ｐ统晒?chuàng)建。在腎癌移植瘤模型實(shí)驗(yàn)中,與對(duì)照組相比,移植瘤的體積和重量經(jīng)過干擾素α2b地處理后明顯被抑制,HE染色檢測結(jié)果顯示:細(xì)胞核與細(xì)胞質(zhì)比例失常,細(xì)胞核明顯增大,細(xì)胞核染色加深,核分裂象明顯異常,腫瘤組織中細(xì)胞無序,大小不一致,形態(tài)不均勻。2.成功分離并純化外泌體,在對(duì)照組與干擾素組中,外泌體的形態(tài)并無明顯差異;與對(duì)照組相比,干擾素α2b能使Renca細(xì)胞分泌的exosome數(shù)量明顯減少。3.在干擾素α2b組中,MDSC與對(duì)照組比較,其所占百分比數(shù)明顯下降。4.在對(duì)照組中,CD4/CD8與干擾素α2b組相比,其所占百分比數(shù)明顯升高。5.在BALB/c小鼠實(shí)驗(yàn)?zāi)Ｐ蛯?shí)驗(yàn)中,與生理鹽水組相比,外泌體組和Renca細(xì)胞組的荷瘤小鼠骨髓中的骨髓源性抑制性細(xì)胞百分比明顯升高。外泌體組與Renca細(xì)胞組相比,荷瘤小鼠骨髓中的骨髓源性抑制性細(xì)胞比例無明顯差異。外泌體組與Renca細(xì)胞組內(nèi),與前10天相比,后20天荷瘤小鼠骨髓中的骨髓源性抑制性細(xì)胞比例明顯升高(P0.05),后20天之間荷瘤小鼠骨髓中的骨髓源性抑制性細(xì)胞比例無明顯變化(P0.05)。NS組其百分比無明顯變化(P0.05)。結(jié)論:Renca細(xì)胞來源的外泌體在小鼠體內(nèi)參與了骨髓源性抑制性細(xì)胞的激活、募集,renca細(xì)胞分泌外泌體被干擾素α2b抑制,從而致使MDSCs的活化、增殖受限,使得移植腫瘤的發(fā)展過程明顯受限。
[Abstract]:Aim: to study the inhibitory effect of interferon-偽-2 b on the growth of renal cell carcinoma in mice.Method 1: 1.The transplanted tumor model of renal cell carcinoma was established and divided into two groups: control group and interferon 偽 2b group, which were treated with blank treatment and interferon 偽 2b intervention respectively.After modeling, the volume and weight of transplanted tumor in two groups of mice were observed and monitored, and the transplanted tumor was removed for HE staining. 2.The exocrine was extracted by ultrafiltration sucrose density gradient centrifugation. The morphological and quantitative changes of exocrine were identified by transmission electron microscope (TEM).In interferon 偽 2b group and control group, lymphocytes were detected by FCM. 4. 4.In the interferon 偽 2b group and the control group, the percentage of myelogenous inhibitory cells was detected by FCM. 5. The BALB / c mouse model was successfully established and divided into normal saline group, exocrine group and Renca cell group.The proportion of bone marrow derived inhibitory cells in the bone marrow of tumor-bearing mice was detected by FCM every 10 days.The result is 1: 1.The transplanted tumor model of renal cell carcinoma and the experimental model of BALB/c mice were successfully established.In the model experiment of transplanted tumor of renal cell carcinoma, compared with the control group, the volume and weight of transplanted tumor were obviously inhibited by HE staining after treatment with interferon 偽 2b. The results showed that the ratio of nucleus to cytoplasm was abnormal and the nucleus increased obviously.The nuclear staining deepened, the mitotic image was obviously abnormal, the cells in the tumor tissue were disordered, the size was not consistent, and the morphology was not uniform. 2.The exocrine was isolated and purified successfully. There was no significant difference in the morphology of exocrine between the control group and the interferon group, and interferon 偽 2b could significantly reduce the number of exosome secreted by Renca cells.The percentage of MDSC in interferon 偽 2b group was significantly lower than that in control group.The percentage of CD4 / CD8 in the control group was significantly higher than that in the interferon 偽 2b group.The percentage of bone marrow-derived inhibitory cells in the bone marrow of tumor bearing mice in the exocrine group and Renca cell group was significantly higher than that in the saline group.There was no significant difference in the proportion of bone marrow-derived inhibitory cells between exocrine group and Renca cell group.In the exocrine group and Renca cell group, compared with the first 10 days,The proportion of bone marrow derived inhibitory cells in the bone marrow of tumor-bearing mice increased significantly after 20 days, while the percentage of bone marrow-derived inhibitory cells in bone marrow of tumor-bearing mice did not change significantly during the latter 20 days. There was no significant change in the percentage of bone marrow-derived inhibitory cells in the P0.05- NS group.Conclusion the exocrine derived from Renca cells is involved in the activation of bone marrow-derived inhibitory cells in mice, and the exocrine secreted by the recruitment of Renca cells is inhibited by interferon 偽 2b, which results in the activation and proliferation of MDSCs is restricted.The development of transplanted tumor is obviously limited.
【學(xué)位授予單位】：重慶醫(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)】：R737.11

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 付必莽;蘇瑩珍;何小文;胡安斌;何曉順;黃潔夫;;逆向解剖法游離股骨/脛骨快捷制備小鼠骨髓細(xì)胞[J];中國組織工程研究與臨床康復(fù);2011年14期

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本文編號(hào)：1760905