本文選題：炎癥 + 免疫調(diào)節(jié)　； 參考：《華中科技大學(xué)》2011年碩士論文

【摘要】：目的:本文擬通過(guò)不同劑量甲氨蝶呤在小鼠炎癥模型中對(duì)一氧化氮的作用來(lái)探討低劑量甲氨蝶呤對(duì)免疫炎癥的影響。 方法:1.用conA尾靜脈注射昆明小鼠,各組分別用不同濃度的MTX處理,48小時(shí)取脾細(xì)胞RNA,檢測(cè)IL-2、Fox-P3的表達(dá);2、用LPS刺激昆明小鼠腹腔巨噬細(xì)胞,各組分別用不同濃度的MTX處理,48小時(shí)后檢測(cè)培養(yǎng)上清NO含量,提取巨噬細(xì)胞RNA,檢測(cè)IL-2、Fox-P3的表達(dá);3、建立conA所致小鼠急性肝炎模型,各組用不同濃度的MTX處理,6小時(shí)后檢測(cè)外周血轉(zhuǎn)氨酶活性;4、建立DH5α大腸桿菌所致小鼠感染模型,各組用不同濃度的MTX處理,6小時(shí)后取肝臟研磨培養(yǎng),24小時(shí)后觀察菌落生長(zhǎng)情況;5、用CFSE標(biāo)記BALB/C小鼠脾臟T細(xì)胞后尾靜脈注入其他BALB/C體內(nèi),并用ConA和不同濃度的MTX處理,48小時(shí)后取脾臟流式細(xì)胞術(shù)檢測(cè)CFSE標(biāo)記的T細(xì)胞增值情況。6、清除小鼠肝臟巨噬細(xì)胞(kupffer cells),36小時(shí)后建立conA所致小鼠急性肝炎模型,6小時(shí)后眼球取血,測(cè)血清中轉(zhuǎn)氨酶活性;7、建立conA所致小鼠急性肝炎模型,用不同濃度的MTX尾靜脈注射,6小時(shí)候分離肝臟Kupper cells,RT-PCR檢測(cè)iNOS的表達(dá)。 結(jié)果:1、低劑量的MTX(0.1ug/ml)可以下調(diào)IL-2的表達(dá)和上調(diào)Fox-P3的表達(dá);2、LPS刺激小鼠腹腔巨噬細(xì)胞后,低劑量的MTX更能有效地抑制巨噬細(xì)胞NO的產(chǎn)生;3、在小鼠肝炎模型中,谷丙轉(zhuǎn)氨酶活性升高,而低劑量MTX處理組轉(zhuǎn)氨酶活性降低,MTX劑量加大后轉(zhuǎn)氨酶活性有所提升。4、在小鼠感染模型中,低劑量的MTX促進(jìn)了細(xì)菌在肝臟的生長(zhǎng),而隨著劑量的加大,這種促進(jìn)作用有所減弱;5、低劑量的MTX沒(méi)有看到明顯的抑制T細(xì)胞增值作用,而相對(duì)高劑量的MTX產(chǎn)生了一定的抑制作用。6、未清除肝臟Kupper cells組轉(zhuǎn)氨酶較清除組明顯升高;7、低劑量的MTX作用后所取的Kupper cells中iNOS表達(dá)較其余組明顯降低。 結(jié)論:MTX劑量的不同其生物學(xué)作用也不同,相對(duì)于高劑量主要表現(xiàn)出細(xì)胞毒性,低劑量的MTX則表現(xiàn)出一定的免疫抑制作用,其通過(guò)抑制巨噬細(xì)胞中iNOS的表達(dá)從而降低在炎癥刺激下巨噬細(xì)胞中NO的產(chǎn)生,達(dá)到其抗炎作用。而在感染模型中,由于其對(duì)免疫功能的抑制,又會(huì)使得感染加重。
[Abstract]:Aim: to investigate the effect of low dose methotrexate on immune inflammation in mouse inflammatory model by using different doses of methotrexate. Method 1: 1. Kunming mice were injected with conA tail vein. Spleen cells were treated with different concentrations of MTX for 48 hours. The expression of IL-2 and Fox-P3 was detected. The macrophages of Kunming mice were stimulated by LPS. Each group was treated with different concentrations of MTX for 48 hours to detect the content of no in culture supernatant, to extract macrophage RNAs, to detect the expression of IL-2 Fox-P3, and to establish a mouse model of acute hepatitis induced by conA. After treated with different concentrations of MTX for 6 hours, the activity of transaminase in peripheral blood of each group was detected and the infection model of mice induced by DH5 偽 Escherichia coli was established. Each group was treated with different concentrations of MTX for 6 hours. After 24 hours of liver grinding culture, the colony growth was observed. The spleen T cells of BALB/C mice were labeled with CFSE and injected into the tail vein of other BALB/C. After 48 hours of treatment with ConA and different concentrations of MTX, splenic T cell proliferation was detected by flow cytometry (FCM). The proliferation of T cells labeled by CFSE was detected by flow cytometry. After 36 hours of clearance of murine hepatic macrophages, the acute hepatitis model induced by conA was established, and the blood was taken from the eyeball after 6 hours. The activity of transaminase in serum was measured to establish mouse model of acute hepatitis induced by conA. The expression of iNOS was detected by RT-PCR of liver Kupper cells at 6 hours after different concentrations of MTX were injected into caudal vein. Results: low dose of MTX 0.1 g / ml could down-regulate the expression of IL-2 and up-regulate the expression of Fox-P3. The low dose of MTX could effectively inhibit the production of no in mouse peritoneal macrophages, and the activity of glutamate-pyruvate aminotransferase increased in mouse hepatitis model. In low dose MTX group, the activity of transaminase increased with the increase of the dose of MTX. In the model of mouse infection, low dose of MTX promoted the growth of bacteria in liver, but with the increase of dose, the activity of transaminase increased. This effect was weakened, and low dose of MTX did not show a significant inhibition of T cell proliferation. However, the relative high dose of MTX produced a certain inhibitory effect. The transaminase expression in the liver Kupper cells group was significantly higher than that in the clearance group, and the iNOS expression in the Kupper cells of the low dose MTX group was significantly lower than that in the other groups. Conclusion the biological effects of different doses of MTX are different. Compared with the high dose, the cytotoxicity is mainly observed, while the low dose of MTX has a certain immunosuppressive effect. By inhibiting the expression of iNOS in macrophages, it can reduce the production of no in macrophages stimulated by inflammation and achieve its anti-inflammatory effect. But in the infection model, because of its inhibition to the immune function, will make the infection aggravation.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R363

【共引文獻(xiàn)】

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1 F. Xiao-Feng Qin;;Dynamic Behavior and Function of Foxp3~+ Regulatory T Cells in Tumor Bearing Host[J];Cellular & Molecular Immunology;2009年01期

2 趙利國(guó);戴敏;李芬;陳正堂;;樹(shù)突狀細(xì)胞在腫瘤中的作用[J];重慶醫(yī)學(xué);2012年11期

3 燕翔;趙曉;焦順昌;孫勝杰;吳亮亮;吳志勇;;晚期肺腺癌患者一線化療后T淋巴細(xì)胞亞群變化及臨床意義[J];中國(guó)肺癌雜志;2012年03期

4 黃香;陳龍邦;;化療藥物的免疫調(diào)節(jié)作用[J];醫(yī)學(xué)研究生學(xué)報(bào);2010年01期

5 鐘翠平;;CD4~+CD25~+調(diào)節(jié)性T細(xì)胞及其功能[J];解剖學(xué)雜志;2008年05期

6 肖琦;蔡軍;肖建生;;調(diào)節(jié)性T細(xì)胞在肝細(xì)胞肝癌中的表達(dá)及化療與免疫治療對(duì)其表達(dá)的影響[J];南昌大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2011年05期

7 祝偉;袁鏗;;CD4~+CD25~+調(diào)節(jié)性T細(xì)胞對(duì)腫瘤免疫的影響[J];南昌大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2011年07期

8 解皓;李海濤;趙彩彥;;CD4~+CD25~+FOXp3~+調(diào)節(jié)性T細(xì)胞參與腫瘤免疫逃逸的研究進(jìn)展[J];臨床薈萃;2009年24期

9 傅艷紅;孫耕耘;;CD_4~+CD_(25)~+調(diào)節(jié)性T細(xì)胞與肺癌的研究進(jìn)展[J];臨床肺科雜志;2011年09期

10 丁江華;龔升平;;節(jié)拍法治療非小細(xì)胞肺癌研究進(jìn)展[J];臨床誤診誤治;2009年12期

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4 馬嵐青;特異性小干擾RNA對(duì)腫瘤細(xì)胞血管內(nèi)皮生長(zhǎng)因子表達(dá)的影響及抗大腸癌血管生成的實(shí)驗(yàn)研究[D];昆明醫(yī)學(xué)院;2008年

5 危華鋒;單鏈抗體介導(dǎo)的小干擾RNA選擇性沉默小鼠調(diào)節(jié)性T細(xì)胞Foxp3基因表達(dá)及效應(yīng)[D];第二軍醫(yī)大學(xué);2008年

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6 劉星;常規(guī)化療聯(lián)合節(jié)拍化療對(duì)乳腺癌裸鼠移植瘤的實(shí)驗(yàn)研究[D];大連醫(yī)科大學(xué);2011年

7 盧曉婷;化療對(duì)乳腺癌患者外周血中Treg細(xì)胞的影響及意義[D];天津醫(yī)科大學(xué);2008年

8 張鐳;抗腫瘤化療對(duì)肺癌患者免疫格局與功能的改變及意義[D];復(fù)旦大學(xué);2008年

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本文編號(hào)：1945372