本文選題：三氯乙烯 + T細(xì)胞��； 參考：《中國(guó)職業(yè)醫(yī)學(xué)》2017年01期

【摘要】：目的研究三氯乙烯(TCE)對(duì)人活化T細(xì)胞的免疫毒性及其作用機(jī)制。方法 1取經(jīng)分化抗原(CD3)和CD28活化的人T細(xì)胞,分別予不同劑量(0.32、0.63、1.25、2.50、5.00、10.00 mmol/L)的TCE染毒,并設(shè)二甲基亞砜(DMSO)組(溶劑對(duì)照組)和對(duì)照組(不予TCE和DMSO處理);培養(yǎng)24 h后,以CCK-8比色法檢測(cè)T細(xì)胞的存活率。2以不同劑量TCE(0.00、2.50和5.00 mmol/L)染毒活化T細(xì)胞24 h后,以流式細(xì)胞術(shù)檢測(cè)細(xì)胞凋亡情況。3取活化T細(xì)胞,分別予不同劑量(0.00、0.32、0.63、1.25、2.50、5.00 mmol/L)的TCE染毒24 h后,以酶聯(lián)免疫吸附實(shí)驗(yàn)檢測(cè)培養(yǎng)上清中白細(xì)胞介素(IL)-2和IL-6等細(xì)胞因子水平。4對(duì)照組、TCE染毒組活化T細(xì)胞分別予劑量為0.00和5.00 mmol/L的TCE染毒,于染毒0、30、60和120 min時(shí)收獲細(xì)胞,以免疫印跡法檢測(cè)信號(hào)傳導(dǎo)蛋白和轉(zhuǎn)錄激活物3(STAT3)及磷酸化STAT3(p-STAT3)的蛋白表達(dá)情況。結(jié)果 1染毒24 h后,10.00 mmol/L TCE染毒組活化T細(xì)胞的存活率分別低于對(duì)照組和DMSO組(P0.05)。2 0.00、2.50和5.00 mmol/L劑量的TCE作用于活化T細(xì)胞時(shí),細(xì)胞凋亡率差異無統(tǒng)計(jì)學(xué)意義(P0.05)。3 0.32、0.63、1.25、2.50和5.00 mmol/L TCE染毒組活化T細(xì)胞培養(yǎng)上清液中IL-2與IL-6水平均高于對(duì)照組(P0.05);隨著TCE染毒劑量的增加,活化T細(xì)胞分泌的IL-2和IL-6水平均上升(P0.01),均呈劑量-效應(yīng)關(guān)系。5個(gè)TCE染毒組活化T細(xì)胞培養(yǎng)上清液中IL-17A、IFN-γ和TGF-β表達(dá)水平分別與對(duì)照組比較,差異均無統(tǒng)計(jì)學(xué)意義(P0.05)。4對(duì)照組活化T細(xì)胞在各個(gè)時(shí)間點(diǎn)p-STAT3蛋白的表達(dá)均較低;TCE染毒組活化T細(xì)胞p-STAT3蛋白表達(dá)在0 min時(shí)間點(diǎn)較低,在30、60和120 min時(shí)間點(diǎn)均上調(diào),且各個(gè)時(shí)間點(diǎn)p-STAT3蛋白表達(dá)均高于對(duì)照組。2組活化T細(xì)胞在各個(gè)時(shí)間點(diǎn)的STAT3總蛋白表達(dá)水平較為一致,且均相對(duì)高于p-STAT3蛋白。結(jié)論 TCE對(duì)活化T細(xì)胞的最大無作用劑量為5.00mmol/L。高劑量TCE(≥10.00 mmol/L)可對(duì)活化T細(xì)胞造成細(xì)胞毒性損傷;低劑量TCE(≤5.00 mmol/L)可刺激活化T細(xì)胞IL-2和IL-6分泌增加;濃度為5.00 mmol/L的TCE可上調(diào)活化T細(xì)胞的STAT3磷酸化水平。
[Abstract]:Objective to study the immunotoxicity of trichloroethylene (TCE) on human activated T cells and its mechanism. Methods 1 Human T cells activated by differentiation antigen (CD3) and CD28 were treated with different doses of TCE of 0.32 ~ 0.63 ~ 1.25 ~ 2.50 ~ 5.00 mmol 路L ~ (-1) and treated with dimethyl sulfoxide (DMSO) group (solvent control group) and control group (TCE and DMSO), respectively, and cultured for 24 h, and then cultured for 24 h, then treated with DMSO (dimethyl sulfoxide) group (solvent control group) and control group (TCE and DMSO). The survival rate of T cells was detected by CCK-8 colorimetry. 2. The survival rate of T cells was measured by CCK-8 colorimetry. The T cells were activated with different doses of TCEC 0.002.50 and 5.00 mmol / L for 24 h. The apoptosis of T cells was detected by flow cytometry. The activated T cells were obtained by flow cytometry. The activated T cells were treated with TCE of 0.000.32 ~ 0.32 ~ (0.32) ~ 1.252.50 mmol / L for 24 h, respectively. Enzyme linked immunosorbent assay (Elisa) was used to detect the levels of cytokines such as interleukin-2 and IL-6 in the supernatant. The activated T cells in the control group were treated with TCE at doses of 0.00 and 5.00 mmol/L, respectively, and the cells were harvested at 30 and 120 min after exposure. The protein expression of signal transduction protein, transcriptional activator 3 (STAT3) and phosphorylated STAT3 (p-STAT3) were detected by Western blotting. Results (1) the survival rate of activated T cells in the 10.00 mmol/L TCE group was lower than that in the control group and DMSO group after 24 hours of exposure. The survival rate of the activated T cells was significantly lower than that of the control group and the DMSO group. The levels of IL-2 and IL-6 in the supernatant of activated T cell culture in the groups exposed to mmol/L TCE were higher than those in the control group, and the levels of IL-2 and IL-6 in the supernatant of activated T cell culture were higher than those in the control group, and with the increase of the dose of TCE, there was no significant difference in apoptosis rate between the two groups. The levels of IL-2 and IL-6 secreted by activated T cells increased in a dose-effect relationship, and the expression levels of IL-17An- 緯 and TGF- 尾 in the supernatants of activated T cells in the five TCE exposed groups were compared with those in the control group, respectively. The expression of p-STAT3 protein in activated T cells of control group was lower at 0 min time point than that in control group at all time points, and increased at 30 ~ 60 and 120 min time points. The expression of p-STAT3 protein at each time point was higher than that of the control group. 2. The expression level of total STAT3 protein in activated T cells at each time point was consistent, and the expression level of p-STAT3 protein was higher than that of p-STAT3 protein. Conclusion the maximum nonactive dose of TCE on activated T cells is 5.00 mmol / L. High dose TCE (鈮，

本文編號(hào)：1809109