本文選題：谷氨酰胺 + 氧化損傷��； 參考：《動(dòng)物營養(yǎng)學(xué)報(bào)》2017年08期

【摘要】：本試驗(yàn)旨在通過研究谷氨酰胺對(duì)過氧化氫(H_2O_2)誘導(dǎo)氧化應(yīng)激人結(jié)腸癌HT-29細(xì)胞損傷和凋亡的影響,闡明谷氨酰胺的抗氧化效果和作用機(jī)理。HT-29細(xì)胞經(jīng)不同濃度[0(對(duì)照組)、0.5、2.0、10.0 mmol/L]谷氨酰胺和0.35 mmol/L H_2O_2分別處理12、24、32 h后,測(cè)定細(xì)胞的超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量,并采用熒光定量PCR方法分析谷氨酰胺對(duì)H_2O_2誘導(dǎo)的細(xì)胞凋亡相關(guān)基因的mRNA相對(duì)表達(dá)量,以及采用膜聯(lián)蛋白V-異硫氰酸熒光素/碘化丙啶雙染法對(duì)HT-29細(xì)胞染色,并用流式細(xì)胞儀檢測(cè)細(xì)胞的凋亡情況。結(jié)果表明:1)處理24 h后,0.5、2.0 mmol/L Gln組SOD活性顯著高于對(duì)照組(P0.05);處理32 h后,0.5、2.0 mmol/L Gln組SOD活性顯著高于對(duì)照組(P0.05),MDA含量顯著低于對(duì)照組(P0.05)。2)處理12 h后,各組天冬氨酸蛋白水解酶-3(Caspase-3)和B淋巴細(xì)胞瘤-2相關(guān)X蛋白(Bax)mRNA相對(duì)表達(dá)量無顯著差異(P0.05)。與對(duì)照組比較,0.5 mmol/L Gln組核轉(zhuǎn)錄因子κB(NF-κB)mRNA相對(duì)表達(dá)量顯著降低(P0.05),0.5與2.0 mmol/L Gln組B淋巴細(xì)胞瘤-2(Bcl-2)mRNA相對(duì)表達(dá)量顯著提高(P0.05)。處理24 h后,與對(duì)照組比較,0.5、2.0和10.0 mmol/L Gln組Caspase-3、NF-κB和Bax mRNA相對(duì)表達(dá)量均顯著降低(P0.05),Bcl-2mRNA相對(duì)表達(dá)量顯著升高(P0.05)。處理32 h后,與對(duì)照組比較,2.0 mmol/L Gln組細(xì)胞表面誘導(dǎo)凋亡分子(FAS)、Caspase-3、NF-κB、Bax mRNA相對(duì)表達(dá)量均顯著降低(P0.05),Bcl-2mRNA相對(duì)表達(dá)量顯著升高(P0.05);10.0 mmol/L Gln組FAS、Caspase-3、NF-κB、Bax mRNA相對(duì)表達(dá)量均顯著升高(P0.05)。3)處理24 h后,與對(duì)照組比較,Gln處理使活細(xì)胞數(shù)量提高了5.32%~11.97%,壞死細(xì)胞數(shù)量降低了6.75%~12.66%。處理32 h后,與對(duì)照組比較,Gln處理使活細(xì)胞數(shù)量提高了1.39%~7.63%,壞死細(xì)胞數(shù)量降低了3.40%~4.57%。由此可見,谷氨酰胺可抑制氧化應(yīng)激反應(yīng),降低H_2O_2誘導(dǎo)的HT-29細(xì)胞凋亡。
[Abstract]:The purpose of this study was to investigate the effects of glutamine on oxidative stress induced HT-29 cell damage and apoptosis in human colon cancer cells induced by H _ 2O _ 2 / H _ 2O _ 2 / H _ 2O _ 2. To elucidate the antioxidation effect and mechanism of glutamine. HT-29 cells were treated with glutamine and 0.35 mmol/L H_2O_2 at different concentration [0 (control group) 0.52.0 ~ 10.0 mmol/L] for 32 h respectively. The activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in HT-29 cells were measured. The relative expression of glutamine on apoptosis-related genes induced by H_2O_2 was analyzed by fluorescence quantitative PCR, and HT-29 cells were stained by fluorescein isothiocyanate / pyridine iodide double staining. Apoptosis was detected by flow cytometry. The results showed that the activity of SOD was significantly higher in 0.5 mmol/L Gln group than that in control group after 24 h treatment, and significantly higher in 0.55 mmol/L Gln group than that in control group after 32 h treatment. There was no significant difference in the relative expression of aspartate proteolytic enzyme-3 (caspase-3) and B lymphocytoma-related X protein (Baxo) mRNA in each group (P 0. 05). Compared with the control group, the relative expression of NF-魏 B)mRNA in the 0.5 mmol/L Gln group was significantly lower than that in the 2. 0 mmol/L Gln group. The relative expression of NF-魏 B)mRNA in the B lymphocytoma of group 2. 0 was significantly higher than that in the group of 2. 0 mmol/L Gln. After treatment for 24 h, the relative expression of Caspase-3 NF- 魏 B and Bax mRNA in the 0. 5 mmol/L Gln group was significantly lower than that in the control group (P 0. 05) and the relative expression of Bcl-2 mRNA was significantly increased (P 0. 05). After treatment for 32 h, the relative expression of apoptosis-inducing Caspase-3NF- 魏 BnBax mRNA was significantly decreased in the 2. 0 mmol/L Gln group compared with the control group. The relative expression of Bcl-2 mRNA in the P0. 05 mmol/L Gln group was significantly higher than that in the control group. The relative expression of Caspase-3 NF- 魏 B Bax mRNA in the P0. 05 mmol/L Gln group was significantly higher than that in the P0. 05 mmol/L Gln group (P 0. 05. 3) after treatment, the relative expression of Caspase-3NF- 魏 B Bax mRNA was significantly increased at 24 h after treatment. Compared with the control group, GLN treatment increased the number of living cells by 5.32 and 11.97, and decreased the number of necrotic cells by 6.75 and 12.66. After 32 hours of treatment, compared with the control group, GLN treatment increased the number of living cells by 1.39 and 7.63, and the number of necrotic cells decreased by 3.40 and 4.57. Therefore, glutamine can inhibit oxidative stress reaction and reduce H_2O_2 induced apoptosis of HT-29 cells.
【作者單位】： 浙江省農(nóng)業(yè)科學(xué)院農(nóng)產(chǎn)品質(zhì)量標(biāo)準(zhǔn)研究所;浙江省植物有害生物防控省部共建國家重點(diǎn)實(shí)驗(yàn)室培育基地;貴州大學(xué)動(dòng)物科學(xué)學(xué)院;浙江大學(xué)動(dòng)物科學(xué)學(xué)院;
【基金】：國家自然科學(xué)基金(31402083) 浙江省自然科學(xué)基金重點(diǎn)項(xiàng)目(LZ15C170001)
【分類號(hào)】：S816

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