本文選題：干擾素αb + 外泌體　； 參考：《重慶醫(yī)科大學(xué)學(xué)報》2017年05期

【摘要】：目的:研究干擾素α2b抑制小鼠腎癌生長及其潛在的作用機制。方法:建立腎癌移植瘤模型,干擾素干預(yù)后將腎癌移植瘤模型分為對照組和干擾素α2b組,監(jiān)測各組小鼠移植瘤體積與重量,切取移植瘤作HE染色,免疫組織化學(xué)檢測移植瘤蛋白BCL-2表達(dá);超濾加蔗糖重水密度梯度離心法提取外泌體,透射電鏡鑒定外泌體形態(tài)及數(shù)量的變化;流式細(xì)胞術(shù)檢測小鼠骨髓中髓源性抑制細(xì)胞(myeloid-derived suppressor cells,MDSCs)比例,小鼠脾臟CD4/CD8T淋巴細(xì)胞比例。建立BALB/c小鼠實驗?zāi)Ｐ?將其分為生理鹽水組、外泌體組和Renca細(xì)胞組,建模后第10、20、30天應(yīng)用流式細(xì)胞儀檢測各組小鼠骨髓MDSCs比例。結(jié)果:成功建立腎癌移植瘤模型與BALB/c小鼠實驗?zāi)Ｐ?在腎癌移植瘤模型實驗中,與對照組比較,干擾素α2b組重量與體積明顯被抑制(P0.01)。HE染色結(jié)果顯示:腫瘤組織中細(xì)胞大小不均,形態(tài)不一,排列紊亂;細(xì)胞核增大,核質(zhì)比例失常,核深染,異常核分裂。免疫組織化學(xué)檢測顯示:對照組為、干擾素α2b組兩組間Bcl-2蛋白陽性表達(dá)率無統(tǒng)計學(xué)差異(P0.05)。成功分離并純化外泌體,對照組、干擾素組2組外泌體形態(tài)并無統(tǒng)計學(xué)差異(P0.05);與對照組比較,干擾素α2b能明顯減少Renca細(xì)胞分泌的外泌體數(shù)量(P0.01)。與對照組比較,干擾素α2b組MDSCs比例明顯降低(P0.05),與對照組相比,干擾素組荷瘤小鼠CD4/CD8T淋巴細(xì)胞比例(P0.05)明顯升高。在BALB/c小鼠實驗?zāi)Ｐ蛯嶒炛?在第10天、20天和30天,與生理鹽水組相比,外泌體組與Renca細(xì)胞組的MDSCs比例明顯增高(P0.01),而外泌體組與Renca細(xì)胞組比較無統(tǒng)計學(xué)差異(P0.05)。外泌體組與Renca細(xì)胞組內(nèi),第20天和30天比第10天要明顯增加MDSCs比例(P0.05),第20天和30天之間MDSCs比例無明顯變化(P0.05)。而生理鹽水組MDSCs比例在第10、20和30天之間比較無統(tǒng)計學(xué)差異(P0.05)。結(jié)論:干擾素α2b通過抑制Renca細(xì)胞分泌外泌體,而Renca細(xì)胞來源外泌體在體內(nèi)參與了小鼠MDSCs的活化、增殖,致使MDSCs的活化、增殖受限,從而抑制小鼠腎癌生長。
[Abstract]:Aim: to study the inhibitory effect of interferon-偽-2b on the growth of renal cell carcinoma in mice and its potential mechanism. Methods: the transplanted tumor model of renal cell carcinoma was established. After interferon intervention, the transplanted tumor model was divided into control group and interferon 偽 2b group. The volume and weight of transplanted tumor in each group were monitored, and the transplanted tumor was removed for HE staining. The expression of BCL-2 was detected by immunohistochemistry, the exocrine was extracted by ultrafiltration and sucrose heavy water density gradient centrifugation, and the morphological and quantitative changes of exocrine were identified by transmission electron microscope (TEM). The ratio of myeloid-derived suppressor cells in bone marrow and the proportion of CD4/CD8T lymphocytes in spleen of mice were detected by flow cytometry. BALB/c mice were divided into three groups: normal saline group, exocrine group and Renca cell group. Flow cytometry was used to detect the proportion of MDSCs in bone marrow of each group. Results: the transplanted tumor model of renal cell carcinoma and the experimental model of BALB/c mice were successfully established. Compared with the control group, the weight and volume of the interferon 偽 2b group were significantly inhibited by P0.01he staining. The results showed that the size of the cells in the tumor tissue was uneven. The nucleus is enlarged, the proportion of nucleus and cytoplasm is abnormal, the nucleus is deep stained and abnormal mitosis. Immunohistochemical examination showed that the positive expression rate of Bcl-2 protein in interferon 偽 2b group was not significantly different from that in interferon 偽 2b group (P 0.05). The exocrine was isolated and purified successfully. There was no significant difference in the morphology of exocrine in control group and interferon group (P 0.05). Compared with control group, interferon 偽 2b could significantly reduce the number of exocrine bodies secreted by Renca cells. Compared with the control group, the proportion of MDSCs in the interferon 偽 2b group was significantly lower than that in the control group. Compared with the control group, the proportion of CD4/CD8T lymphocytes in the interferon group was significantly higher than that in the control group. In the BALB/c mouse model experiment, on the 10th day, 20 days and 30 days, compared with the normal saline group, the proportion of MDSCs in the exocrine group and Renca cell group was significantly higher than that in the control group, but there was no significant difference between the exocrine body group and the Renca cell group. In the exocrine group and the Renca cell group, the proportion of MDSCs was significantly increased on the 20th and 30th days as compared with the 10th day, but there was no significant change in the MDSCs ratio between the 20th and the 30th day. However, there was no significant difference in the proportion of MDSCs between the 10 ~ (th) day and the 30 ~ (th) day in the saline group (P 0.05). Conclusion: interferon 偽 2b inhibits the exocrine secretion of Renca cells, and the exocrine derived from Renca cells participate in the activation and proliferation of mouse MDSCs in vivo, resulting in the limited activation and proliferation of MDSCs, thus inhibiting the growth of mouse renal cell carcinoma.
【作者單位】： 重慶醫(yī)科大學(xué)附屬第一醫(yī)院泌尿外科;重慶醫(yī)科大學(xué)檢驗醫(yī)學(xué)院;
【基金】：國家自然科學(xué)基金資助項目(編號:81272572) 重慶市衛(wèi)生局科研資助項目(編號:2012242)
【分類號】：R737.11

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