發(fā)布時間：2018-05-08 22:38

  本文選題：鉬 + 鎘��； 參考：《江西農(nóng)業(yè)大學》2017年碩士論文

【摘要】：為研究鉬鎘聯(lián)合誘導對鴨腎小管上皮細胞氧化損傷的影響。本實驗通過篩網(wǎng)過濾法和酶消化法建立鴨腎小管上皮細胞體外培養(yǎng)模型,(NH4)6Mo7O24·4H2O和3CdSO4·8H2O分別作為鉬源和鎘源,用堿性磷酸酶化學染色(改良鈣鈷法)、MTT法、冠式法分別對細胞進行鑒定、測定腎小管上皮細胞活性和鉬鎘單獨染毒12 h的IC50。對傳一代腎小管上皮細胞進行鉬(480μmol/LMo、960μmol/LMo)、鎘(2.5μmol/LCd、5μmol/LCd)及其聯(lián)合(480μmol/L Mo+2.5μmol/L Cd、960μmol/L Mo+5μmol/L Cd)染毒,測定染毒3 h、6 h、12 h、24 h細胞存活率、染毒12 h LDH釋放率和SOD活性、T-AOC能力和GSH、MDA含量;并進一步測定NAC對染毒后細胞存活率和LDH釋放率的影響。結(jié)果表明:(1)堿性磷酸酶化學染色顯示,陽性細胞被染為黑色而陰性對照細胞不被染色,對陽性細胞進行計數(shù),顯示近端腎小管上皮細胞純度為95%以上。(2)原代細胞生長曲線:24 h前處于生長期,24 h~88 h處于對數(shù)期且78h細胞活性最強,90 h進入平穩(wěn)期。傳一代細胞生長曲線:12 h前處于生長期,12 h~70 h處于對數(shù)期且56 h細胞活性最強,70 h進入平穩(wěn)期。(3)(NH4)6Mo7O24·4H2O和3CdSO4·8H2O染毒12 h的IC50分別為1773.64μmol/LMo、24.17μmol/LCd。(4)細胞存活率:高劑量鉬組、鎘組及其聯(lián)合組從3 h開始,極顯著低于對照組(P0.01);低劑量鉬組從6 h開始,顯著低于對照組(P0.05),且呈劑量-時間效應(yīng);同一染毒時間,各聯(lián)合染毒組的細胞存活率均低于各相關(guān)單獨染毒組;析因分析表明,從染毒12 h開始鉬鎘之間存在交互作用(P0.05)。NAC可以提高染毒組細胞的存活率(P0.01)。(5)細胞內(nèi)LDH釋放率:各染毒組與對照組相比極顯著升高(P0.01),且鉬鎘聯(lián)合染毒組高于單獨染毒組,NAC對降低LDH釋放率無明顯作用(P0.05)。(6)SOD活性、T-AOC能力及GSH含量:染毒組均顯著或極顯著低于對照組(P0.05或P0.01);MDA含量:染毒組均極顯著高于對照組(P0.01)。結(jié)論:鉬鎘及其聯(lián)合染毒對鴨腎小管上皮細胞的氧化損傷呈劑量-時間效應(yīng),鉬鎘呈協(xié)同效應(yīng);NAC對鉬鎘及其聯(lián)合所致的腎小管上皮細胞毒性損傷有一定的保護作用。
[Abstract]:To study the effect of molybdenum and cadmium combined with cadmium on oxidative damage of duck renal tubular epithelial cells. In this experiment, the model of duck renal tubular epithelial cell culture in vitro was established by screening filter method and enzyme digestion method. 4H2O and 3CdSO4 8H2O were used as molybdenum source and cadmium source respectively. The cells were identified by alkaline phosphatase staining (modified Calcium-Cobalt method, Coronal method, respectively). The activity of renal tubular epithelial cells and IC50 induced by molybdenum and cadmium alone for 12 h were measured. A new generation of renal tubular epithelial cells was treated with Mo-480 渭 mol / L Mo-960 渭 mol 路L ~ (-1), CD ~ (2. 5) 渭 mol / L ~ (5 渭 mol 路L ~ (L) and combined with 480 渭 mol/L Mo ~ (2.5 渭 mol/L) CD ~ (6) mol/L Mo ~ (9) 渭 mol/L Mo ~ (5 渭 mol/L). The cell survival rate, the LDH release rate and the activity of SOD, T-AOC and GSH-MDA content were measured at 3 h, 6 h, 12 h and 24 h after exposure. The effects of NAC on cell survival rate and LDH release rate were further determined. The results showed that the positive cells were stained black but the negative control cells were not stained, and the positive cells were counted. The results showed that the purity of proximal tubular epithelial cells was more than 95%.) the growth curve of primary cells was in the growth phase at 24 h and in logarithmic phase at 88 h before 0. 24 h, and the highest cell activity at 78 h was in stable phase at 90 h. The growth curve of the first generation of cells was in the growth phase for 12 h and in the logarithmic phase for 70 h, and the cell activity of 56 h was the highest in the stationary phase. The IC50 of NH4Mo6Mo7O24 4H2O and 3CdSO4 8H2O exposed to 3CdSO4 8H2O for 12 h were 1773.64 渭 mol / L Mo-24.17 渭 mol / L LCd.m4) cell survival rate: high dose molybdenum group, the survival rate of the cells was 1773.64 渭 mol / L Moo 24.17 渭 mol / L LCd.O4, respectively. The cadmium group and its combined group were significantly lower than the control group (P 0.01) from 3 h, and the low dose molybdenum group from 6 h, significantly lower than the control group (P 0.05), and showed a dose-time effect. The cell survival rate of each combined exposure group was lower than that of each other. The interaction between molybdenum and cadmium (P0.05N. NAC) could increase the survival rate of cells and the release rate of LDH in the cells of the exposed group was significantly higher than that of the control group, and the combination of molybdenum and cadmium group was higher than that of the control group. There was no significant effect of NAC on reducing the release rate of LDH. The activity of P0.05N. 6C and the activity of T-AOC and the content of GSH were significantly or extremely lower in the exposed group than those in the control group (P0.05 or P0.01). The content of LDH in the exposed group was significantly higher than that in the control group (P 0.01). Conclusion: the oxidative damage of duckling renal tubular epithelial cells induced by molybdenum and cadmium combined with NAC has a dose-time effect and a synergistic effect of molybdenum and cadmium. NAC has a certain protective effect on the toxicity of renal tubular epithelial cells induced by molybdenum and cadmium.
【學位授予單位】：江西農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S859.8

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