【摘要】：目的:觀察氧糖剝奪(oxygen-glucose deprivation,OGD)后,神經(jīng)膠質(zhì)瘤細(xì)胞U251的乳酸轉(zhuǎn)運(yùn)和單羧酸轉(zhuǎn)運(yùn)蛋白(monocarboxylate transporter,MCT)1、4的表達(dá)變化,以探討膠質(zhì)瘤增殖生長(zhǎng)的分子機(jī)制。方法:U251細(xì)胞隨機(jī)分為對(duì)照組和模型組(OGD組),模型組的干預(yù)時(shí)間點(diǎn)為1、3、6 h。四甲基偶氮唑鹽比色法(methyl thiazolyl tetrazolium,MTT)檢測(cè)細(xì)胞增殖生長(zhǎng)情況,HE染色觀察細(xì)胞的形態(tài)變化,乳酸試劑盒檢測(cè)細(xì)胞培養(yǎng)液中的乳酸含量,免疫熒光和免疫印跡觀察MCT1和MCT4的表達(dá)變化情況。結(jié)果:與對(duì)照組相比,隨著氧糖剝奪時(shí)間延長(zhǎng),模型組細(xì)胞增殖能力下降(對(duì)照組、OGD 1 h、OGD 3 h、OGD 6 h組的光密度值分別為0.584±0.031、0.537±0.046、0.441±0.079和0.200±0.011),細(xì)胞培養(yǎng)液中的乳酸濃度增高(對(duì)照組、OGD 1 h、OGD 3 h、OGD 6 h組乳酸濃度值分別為0.150±0.029、0.707±0.022、1.167±0.024和1.325±0.023)。MCT1、4的表達(dá)上調(diào),并隨氧糖剝奪時(shí)間延長(zhǎng)其表達(dá)逐漸增強(qiáng)(MCT1:F=46.256,P=0.000;MCT4:F=247.400,P=0.000)。細(xì)胞培養(yǎng)液中加入MCTs的抑制劑4-CIN后,其乳酸濃度降低(對(duì)照組、150μmol/L組、425μmol/L組的乳酸濃度值分別為0.249±0.012、0.222±0.016、0.167±0.117)。結(jié)論:U251細(xì)胞氧糖剝奪后,隨著細(xì)胞培養(yǎng)液中乳酸濃度增加,MCT1、4亦表達(dá)上調(diào),而細(xì)胞的增殖生長(zhǎng)能力下降;抑制MCT的表達(dá),細(xì)胞培養(yǎng)液中的乳酸濃度降低,提示MCT1、4可能通過(guò)參與神經(jīng)膠質(zhì)瘤缺血缺氧微環(huán)境中乳酸的轉(zhuǎn)運(yùn),進(jìn)而調(diào)控細(xì)胞的增殖生長(zhǎng)過(guò)程。
[Abstract]:Aim: to investigate the changes of lactate transport and monocarboxylate transporter (monocarboxylate transporter,MCT) 1 / 4 expression in glioma cell line U251 after oxygen glucose deprivation (oxygen-glucose deprivation,OGD) in order to explore the molecular mechanism of glioma proliferation and growth. Methods: U251 cells were randomly divided into control group and model group (OGD group). (methyl thiazolyl tetrazolium,MTT) was used to detect cell proliferation and growth, HE staining was used to observe the morphological changes of cells, and lactic acid kit was used to detect the content of lactic acid in cell culture medium. Immunofluorescence and Western blot were used to observe the expression of MCT1 and MCT4. Results: compared with the control group, the proliferation ability of the model group decreased with the prolongation of oxygen glucose deprivation time (control group, OGD 1 h, OGD 3 h). The optical density of OGD for 6 h was 0.584 鹵0.031 鹵0.046 鹵0.441 鹵0.079 and 0.200 鹵0.011, respectively. The concentration of lactic acid in cell culture medium was increased (control group, OGD 1 h, OGD 3 h). The concentration of lactic acid in OGD 6 h group was 0.150 鹵0.029 鹵0.707 鹵0.022 鹵1.167 鹵0.024 and 1.325 鹵0.023, respectively. The expression of MCT1,4 was up-regulated and gradually increased with the prolongation of oxygen glucose deprivation time (MCT1:F=46.256,P=0.000;). MCT4:F=247.400,P=0.000) The concentration of lactic acid decreased after the addition of 4-CIN, an inhibitor of MCTs (control group, 150 渭 mol/L group, 425 渭 mol/L group), the lactate concentration was 0.249 鹵0.012 鹵0.222 鹵0.016 渭 mol/L 0.167 鹵0.117, respectively. Conclusion: after U251 cells were deprived of oxygen sugar, the expression of MCT1,4 was up-regulated with the increase of lactic acid concentration in the culture medium, but the proliferation and growth ability of U251 cells decreased. The inhibition of MCT expression and the decrease of lactic acid concentration in cell culture medium suggest that MCT1,4 may regulate the proliferation and growth of glioma cells by participating in the transport of lactic acid in hypoxia-ischemia microenvironment.
【作者單位】： 重慶醫(yī)科大學(xué)神經(jīng)科學(xué)中心重慶醫(yī)科大學(xué)基礎(chǔ)醫(yī)學(xué)院解剖教研室;
【基金】：國(guó)家自然科學(xué)基金資助項(xiàng)目(編號(hào):81671312、81000566) 重慶市基礎(chǔ)與前沿研究計(jì)劃資助項(xiàng)目(編號(hào):cstc2015jcyj A10018)
【分類(lèi)號(hào)】：R739.41

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