【摘要】：【目的】研究錳對(duì)大鼠原代小膠質(zhì)細(xì)胞氧化損傷和炎癥因子白細(xì)胞介素(IL-1β、IL-6)、腫瘤壞死因子(TNF-α)、前列腺素2(PGE2)表達(dá)的影響,探討對(duì)氨基水楊酸鈉(PAS-Na)對(duì)錳導(dǎo)致的小膠質(zhì)細(xì)胞損傷的干預(yù)作用�！痉椒ā�(1)原代培養(yǎng)的小膠質(zhì)細(xì)胞經(jīng)過分離提純和鑒定后,(1)錳毒性試驗(yàn):種于96孔板中的細(xì)胞分別給予0、200、300、400、500μmol/L濃度MnCl2處理24h。(2)PAS-Na無毒篩選試驗(yàn):孔接種于96孔板中的細(xì)胞分別給予0、50、150、450μmol/L濃度PAS-Na處理24h。(3)PAS-Na干預(yù)試驗(yàn):小膠質(zhì)細(xì)胞被隨機(jī)分為對(duì)照組、染錳組、PAS對(duì)照組、50、150、450-PAS干預(yù)組,其中對(duì)照組、染錳組、PAS對(duì)照組給予正常DMEM完全培養(yǎng)基,50、150、450-PAS干預(yù)組給予400μmol/L濃度MnCl2處理24h后,對(duì)照組給予正常DMEM完全培養(yǎng)基換液,染錳組給予400μmol/L濃度MnCl2處理24h,PAS對(duì)照組給予450μmol/L濃度PAS-Na處理24h,50、150、450-PAS干預(yù)組分別加入50、150、450μmol/L濃度PAS-Na處理24h。(2)用MTT試劑測(cè)定小膠質(zhì)細(xì)胞存活率,DCFH-DA探針標(biāo)記小膠質(zhì)細(xì)胞氧化損傷情況,酶聯(lián)免疫吸附法測(cè)定細(xì)胞上清液IL-1β、IL-6、TNF-α、PGE2炎癥因子含量,熒光定量PCR檢測(cè)小膠質(zhì)細(xì)胞IL-1β、IL-6、TNF-αmRNA表達(dá)�！窘Y(jié)果】細(xì)胞存活率檢測(cè):低濃度的MnCl2能刺激原代小膠質(zhì)細(xì)胞的增殖存活率增加,高濃度MnCl2對(duì)小膠質(zhì)細(xì)胞有一定的細(xì)胞毒性,MnCl2處理24h后小膠質(zhì)細(xì)胞存活率在400μmol/L和500μmol/L組比對(duì)照組低(P0.05)。PAS-Na處理24h后,與對(duì)照組比較,各劑量處理組細(xì)胞形態(tài)和存活率沒有明顯變化(P0.05)。染Mn 24h,50、150、450μmol/L濃度PAS干預(yù)24h后,染Mn組細(xì)胞存活率比對(duì)照組低(P0.05)。與染Mn組比較,50、150、450-PAS干預(yù)組小膠質(zhì)細(xì)胞存活率升高,差異有統(tǒng)計(jì)學(xué)差異(P0.05)。細(xì)胞內(nèi)活性氧DCFH-DA標(biāo)記檢驗(yàn):發(fā)現(xiàn)Mn處理可以顯著增加細(xì)胞活性氧的生成,50、150、450-PAS干預(yù)組細(xì)胞活性氧生成比染Mn組少。細(xì)胞上清炎癥因子:與對(duì)照組相比,MnCl2使小膠質(zhì)細(xì)胞上清液中IL-1、TNFα分泌增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。50、150、450-PAS干預(yù)組細(xì)胞TNFα表達(dá)低于染Mn組(P0.05)。小膠質(zhì)細(xì)胞IL-1β、TNF-αmRNA表達(dá):與對(duì)照組相比,染Mn組IL-1β、TNFαmRNA表達(dá)量增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。50、150、450-PAS干預(yù)組小膠質(zhì)細(xì)胞IL-1βmRNA表達(dá)低于染Mn組(P0.05)。150、450-PAS干預(yù)組小膠質(zhì)細(xì)胞TNF-αmRNA表達(dá)低于染Mn組(P0.05)�！窘Y(jié)論】(1)過量錳暴露會(huì)對(duì)原代小膠質(zhì)細(xì)胞造成損傷,導(dǎo)致細(xì)胞內(nèi)活性氧生成,IL-1β、TNF-αmRNA表達(dá)量升高,其對(duì)應(yīng)的炎癥因子IL-1β、TNF-α分泌增加。(2)PAS-Na對(duì)錳導(dǎo)致的小膠質(zhì)細(xì)胞損傷具有一定的干預(yù)作用,這可能與PAS-Na的抗氧化和抗炎作用有關(guān)。
[Abstract]:[objective] to study the effects of manganese on oxidative injury of rat microglia and the expression of IL-1 尾 -IL-6, TNF- 偽 and PGE2. To investigate the effect of sodium p-aminosalicylate (PAS-Na) on the damage of microglia induced by manganese. [methods] (1) the primary cultured microglia were purified and identified. (1) Manganese toxicity test: cell division in 96-well plate. Don't be treated with 0 200300400500 渭 mol/L MnCl2 for 24 h. (2) PAS-Na nontoxic screening test: cells inoculated in 96-well plate were treated with 0 50150450 渭 mol/L PAS-Na for 24 h. (3) PAS-Na intervention test: microglial cells were randomly divided into control group. The rats in the control group were treated with normal DMEM complete medium 50150450-pas for 24 h, and the control group was treated with MnCl2 at the concentration of 400 渭 mol/L for 24 hours. The control group was treated with the normal DMEM complete medium for 24 hours, and the control group was treated with the normal DMEM complete medium for 24 hours, and the control group was treated with the normal DMEM complete medium for 24 hours. The manganese exposed group was treated with 400 渭 mol/L MnCl2 for 24 h, the control group was treated with 450 渭 mol/L PAS-Na for 24 h, the control group was treated with 50150450 渭 mol/L PAS-Na for 24 h. (2) the survival rate of microglia was measured by MTT reagent and the oxidative damage of microglia was labeled with DCFH-DA probe. Enzyme linked immunosorbent assay (Elisa) was used to detect the content of IL-1 尾 -TNF- 偽 PGE2 in supernatant, and fluorescence quantitative PCR was used to detect the expression of IL-1 尾 -IL-6TNF- 偽 mRNA in microglia. [results] Cell survival rate: low concentration of MnCl2 could stimulate the proliferation and survival rate of primary microglia. The survival rate of microglia in the 400 渭 mol/L and 500 渭 mol/L groups was lower than that in the control group (P0.05). PAS-Na had no significant changes in cell morphology and survival rate compared with the control group (P0.05). The cell survival rate of the Mn group was lower than that of the control group (P0.05) after 24 h PAS treatment with Mn 50150450 渭 mol/L for 24 h. Compared with the Mn group, the survival rate of microglia in the 50 150450-pas group was higher than that in the control group (P0.05). DCFH-DA labeling test of intracellular reactive oxygen species: it was found that Mn treatment could significantly increase the production of reactive oxygen species in cells treated with 50150450-pas, which was less than that in Mn infected group. The expression of IL-1,TNF 偽 in the supernatant of microglia was significantly higher than that in the control group (P0.05). The expression of TNF 偽 in the cells of the intervention group was lower than that in the Mn group (P0.05). The expression of IL-1 尾 -TNF- 偽 mRNA in microglial cells: compared with the control group, the expression of IL-1 尾 -TNF- 偽 mRNA in Mn group was higher than that in control group. The difference was statistically significant (P0.05) .50150450-pas intervention group was lower than Mn group (P0.05) .150450-pas intervention group microglial cell TNF- 偽 mRNA expression was lower than that of Mn group (P0.05). [conclusion] (1) excessive manganese exposure will cause damage to primary microglia cells. The expression of IL-1 尾 -TNF- 偽 mRNA increased, and the corresponding inflammatory factor IL-1 尾 -TNF- 偽 secretion increased. (2) PAS-Na may interfere with the damage of microglia induced by manganese, which may be related to the antioxidant and anti-inflammatory effects of PAS-Na.
【學(xué)位授予單位】：廣西醫(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)】：R114

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 張瓊;劉文娟;曹霞;;小膠質(zhì)細(xì)胞特性及其功能的研究進(jìn)展[J];醫(yī)學(xué)研究生學(xué)報(bào);2017年02期

2 王偉;趙全軍;劉爽;王濤;史鐵鈞;王淑為;郭欣茹;;大鼠腦損傷海水浸泡后損傷腦組織小膠質(zhì)細(xì)胞炎性改變[J];中國(guó)神經(jīng)免疫學(xué)和神經(jīng)病學(xué)雜志;2016年06期

3 莫瑞康;王進(jìn);黃堅(jiān)毅;文巧林;黃錦鳳;;腦內(nèi)注射染錳對(duì)大鼠行為學(xué)及黑質(zhì)多巴胺能神經(jīng)元的影響[J];中華勞動(dòng)衛(wèi)生職業(yè)病雜志;2016年06期

4 褚慧;王乾興;;錳的雄性生殖毒理效應(yīng)及其機(jī)制研究進(jìn)展[J];現(xiàn)代醫(yī)藥衛(wèi)生;2016年07期

5 張瓊;劉文娟;曹霞;;小膠質(zhì)細(xì)胞對(duì)神經(jīng)元作用的研究進(jìn)展[J];醫(yī)學(xué)研究生學(xué)報(bào);2016年04期

6 官瑞麗;王濤;陳景元;駱文靜;劉明朝;;鉛錳聯(lián)合暴露通過誘導(dǎo)小膠質(zhì)細(xì)胞活化引起星形膠質(zhì)細(xì)胞活化及谷氨酰胺合成酶的活性降低[J];細(xì)胞與分子免疫學(xué)雜志;2016年03期

7 袁宗祥;莫玉煥;李少軍;區(qū)仕燕;姜岳明;陳海濱;;PAS-Na對(duì)染錳大鼠血清脂質(zhì)過氧化、血和骨骼金屬元素水平的影響[J];毒理學(xué)雜志;2015年04期

8 Howe CL;Kaptzan T;Magaa SM;Ayers-Ringler JR;LaFrance-Corey RG;Lucchinetti CF;;視神經(jīng)脊髓炎IgG刺激原代培養(yǎng)大鼠星形膠質(zhì)細(xì)胞免疫反應(yīng)[J];神經(jīng)損傷與功能重建;2014年03期

9 毛葉挺;;錳對(duì)接觸工人神經(jīng)和心血管系統(tǒng)的影響[J];中國(guó)工業(yè)醫(yī)學(xué)雜志;2013年02期

10 張星虎;;多發(fā)性硬化的診斷[J];中國(guó)神經(jīng)免疫學(xué)和神經(jīng)病學(xué)雜志;2013年02期

，

本文編號(hào)：2200760