牛磺酸对缺氧所致神经胶质细胞凋亡的抑制作用
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摘要
目的探讨牛磺酸对缺氧引起的神经胶质细胞凋亡的抑制作用方法原代培养神经胶质细胞并制作细胞缺氧模型,实验分为空白对照组、缺氧模型组、缺氧+牛磺酸组。细胞缺氧6 h后加入牛磺酸3 mmol/L继续培养24 h和48 h。采用Annexin V-FITC/PI双染色流式细胞术检测细胞凋亡率,RT-PCR检测凋亡相关基因表达水平。结果缺氧组48 h早期凋亡率为(2.48±0.92)%,中晚期凋亡率为(2.97±0.75)%,均显著高于对照组的(1.02±0.44)%(P<0.05);牛磺酸48 h组早期凋亡率为(1.39±1.03)%,中晚期凋亡率为(1.62±0.1)%,均显著低于缺氧组(P<0.05)。缺氧组Bax mRNA相对表达量均显著高于对照组和牛磺酸组(P<0.01);牛磺酸48 h组Bcl-2 mRNA相对表达量比缺氧组升高2倍(P<0.01);牛磺酸组HIF-1αmRNA表达量均比缺氧组升高显著(P<0.01)。结论牛磺酸有助于抑制缺氧所致的神经胶质细胞早期和中晚期凋亡率。
Objective To study the anti-apoptosis effect of taurine on hypoxic glial cells. Methods The glial cells were primarily cultured and divided into three groups,the control group,hypoxia group,and hypoxia + taurine group. The hypoxia model was established by putting cells in a tank filled with oxygen-free mixed gas( 95% N2,5%CO2) with oxygen concentration less than 1%. When the hypoxia time( 6 hours) was finished,taurine( 3 mmol / L) was added to incubate for hypoxia + taurine group. Finally,the glial cells of all groups were collected for FCM and RT-PCR tests after 24 and 48 hours,respectively. Results The earlier apoptosis rate was( 2. 48 ± 0. 92) %,the middle-late apoptosis rate was( 2. 97 ± 0. 75) % for 48 h hypoxia group, they were significantly higher than the control group( 1. 02 ± 0. 44) %( P < 0. 05). The earlier apoptosis rate( 1. 39 ± 1. 03) % and middle-late apoptosis rate( 1. 62 ± 0. 1) % for 48 h taurine group were significantly lower than the hypoxia group( P < 0. 05). The relative expression level of Bax mRNA for hypoxia group increased by five times higher than the control group,and two times than the hypoxia + taurine group( P < 0. 05),and that of Bcl-2 mRNA for 48 h hypoxia + taurine group was two times higher than the hypoxia group( P < 0. 01). The relative expression levels of HIF-1α mRNA for 24 h and 48 h hypoxia +taurine groups were significantly higher than the hypoxia group( P < 0. 01). Conclusion Taurine could obviously inhibited the apoptosis of glial cells induced by hypoxia.
Objective To study the anti-apoptosis effect of taurine on hypoxic glial cells. Methods The glial cells were primarily cultured and divided into three groups,the control group,hypoxia group,and hypoxia + taurine group. The hypoxia model was established by putting cells in a tank filled with oxygen-free mixed gas( 95% N2,5%CO2) with oxygen concentration less than 1%. When the hypoxia time( 6 hours) was finished,taurine( 3 mmol / L) was added to incubate for hypoxia + taurine group. Finally,the glial cells of all groups were collected for FCM and RT-PCR tests after 24 and 48 hours,respectively. Results The earlier apoptosis rate was( 2. 48 ± 0. 92) %,the middle-late apoptosis rate was( 2. 97 ± 0. 75) % for 48 h hypoxia group, they were significantly higher than the control group( 1. 02 ± 0. 44) %( P < 0. 05). The earlier apoptosis rate( 1. 39 ± 1. 03) % and middle-late apoptosis rate( 1. 62 ± 0. 1) % for 48 h taurine group were significantly lower than the hypoxia group( P < 0. 05). The relative expression level of Bax mRNA for hypoxia group increased by five times higher than the control group,and two times than the hypoxia + taurine group( P < 0. 05),and that of Bcl-2 mRNA for 48 h hypoxia + taurine group was two times higher than the hypoxia group( P < 0. 01). The relative expression levels of HIF-1α mRNA for 24 h and 48 h hypoxia +taurine groups were significantly higher than the hypoxia group( P < 0. 01). Conclusion Taurine could obviously inhibited the apoptosis of glial cells induced by hypoxia.
引文
[1]CHEN K,ZHANG Q,WANG J,et al.Taurine protects transformed rat retinal ganglion cells from hypoxia-induced apoptosis by preventing mitochondrial dysfunction[J].Brain Res,2009(1279):131-138.
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[5]ZHANG X,LU H,WANG Y,et al.Taurine induces the apoptosis of breast cancer cells by regulating apoptosis-related proteins of mitochondria[J].Int J Mol Med,2015,35(1):218-226.
[6]常小丽,黄慧玲,王琴,等.牛磺酸对大鼠脑创伤凝血状态影响的研究[J].创伤外科杂志,2010,12(4):355-357.
[7]莫丽冬,黄慧玲,张学斌,等.牛磺酸对急性重型颅脑创伤大鼠脑血流的影响[J].山东医药,2012,52(9):4-6.
[8]LIU J,WANG X F,LIU Y,et al.Antenatal taurine reduces cerebral cell apoptosis in fetal rats with intrauterine growth restriction[J].Neural Regen Res,2013,8(23):2190-2197.
[9]SETYARANI M,ZINELLU A,CARRU C,et al.High dietary taurine inhibits myocardial apoptosis during an atherogenic diet:association with increased myocardial HSP70 and HSF-1 but not caspase 3[J].Eur J Nutr,2014,53(3):929-937.
[2]王琴,黄慧玲.牛磺酸对神经细胞损伤保护作用的研究[J].国际检验医学杂志,2008,29(5):443-445.
[3]ZENG K,XU H,MI M,et al.Dietary taurine supplementation prevents glial alterations in retina of diabetic rats[J].Neurochem Res,2009,34(2):244-254.
[4]ZENG K,MING J,YANG N,et al.Taurine prevents high glucose-induced angiopoietin-2/tie-2 system alterations and apoptosis in retinal microvascular pericytes[J].Mol Cell Biochem,2014,396(1-2):239-248.
[5]ZHANG X,LU H,WANG Y,et al.Taurine induces the apoptosis of breast cancer cells by regulating apoptosis-related proteins of mitochondria[J].Int J Mol Med,2015,35(1):218-226.
[6]常小丽,黄慧玲,王琴,等.牛磺酸对大鼠脑创伤凝血状态影响的研究[J].创伤外科杂志,2010,12(4):355-357.
[7]莫丽冬,黄慧玲,张学斌,等.牛磺酸对急性重型颅脑创伤大鼠脑血流的影响[J].山东医药,2012,52(9):4-6.
[8]LIU J,WANG X F,LIU Y,et al.Antenatal taurine reduces cerebral cell apoptosis in fetal rats with intrauterine growth restriction[J].Neural Regen Res,2013,8(23):2190-2197.
[9]SETYARANI M,ZINELLU A,CARRU C,et al.High dietary taurine inhibits myocardial apoptosis during an atherogenic diet:association with increased myocardial HSP70 and HSF-1 but not caspase 3[J].Eur J Nutr,2014,53(3):929-937.