Bcl-2在染氟大鼠肾小管上皮细胞氧化应激态中的表达  

Expression of Bcl-2 on the oxidative stress of renal tubular cells treated by NaF

作　　者：徐辉[1] 张静敏[1] 常明[1] 崔伦文[1] 李广生[1] 

机构地区：[1]吉林大学医学院再生医学科学研究所,长春130021

出　　处：《中国地方病学杂志》2005年第1期17-20,共4页Chinese Jouranl of Endemiology

基　　金：国家自然科学基金资助项目(39730390)

摘　　要：目的观察不同染氟(NaF,F-)条件下大鼠肾小管上皮细胞氧化应激能力变化及Bcl-2在氧化应激时的表达。方法采用体外原代细胞培养方法,相差显微镜观察大鼠肾小管上皮细胞形态结构变化;测定大鼠肾小管上皮细胞在染氟(F-)0.050、0.250、0.750、1.250mmol/L和不同氟暴露时间条件下,肾小管上皮细胞的增殖活性和丙二醛(MDA)水平、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性;利用半定量RT-PCR方法分析大鼠肾小管上皮细胞在氟暴露0.050～0.625mmol/L48hBcl-2的表达。结果相差显微镜下,染氟0.750、1.250mmol/L组部分肾小管上皮细胞缩为圆形和有细胞脱落现象。在24h,0.005mmol/L组(0.48±0.10)、0.025mmol/L组(0.46±0.12)、0.250mmol/L组(0.36±0.04)肾小管上皮细胞的增殖活性较对照组(0.29±0.09)有不同程度升高,但在72h,1.250mmol/L组(0.16±0.02)细胞的增殖活性明显降低。肾小管上皮细胞MDA水平在染氟48h内,0.25mmol/L组(626.15±124.02)、0.75mmol/L组(498.74±119.88)、1.250mmol/L组(441.99±53.57)与对照组(232.62±21.00)比较,升高显著;SOD酶活性在48h内,随着染氟量的增加而有升高,在0.750mmol/L组(1298.66±248.53)、1.250mmol/L组(1664.70±277.68),酶活性明显高于对照组(741.67±265.56);CAT酶活性,在0.Objective To investigate the role of oxidative stress on renal injury induced by fluoride and the expression of Bcl-2 on the oxidative stress of renal tubular cells exposed to sodium fluoride (NaF,F-) at different concentration. Methods The renal tubular cells were cultured in vitro and treated by NaF(F-). The MTT method and the level of malondialdehyde (MDA), superoxide dismutase (SOD), catalase(CAT) were tested by biochemical techniques. The expression of Bcl-2 was studied by reverse transcription polymerase chain reaction (RT-PCR). Results It was observed that renal tubular cells fell off in the 0.750, 1.250 mmol/L group. The proliferating state dramatically intensified in 0.005 mmol/L group (0.48 ± 0.10), 0.025 mol/L group (0.46 ± 0.12),0.250 mmol/L group (0.36 ± 0.04) compared to the control (0.29 ± 0.09), cells proliferation obviously decreased in the 1.250 mmol/L group (0.16 ± 0.02). The level of MDA in cells exposed to fluoride 48 h was markedly increased in 0.250 mmol /L group (626.15 ± 124.02)?0.75 mmol /L group(498.74 ± 119.88)?1.250 mmol/L group(441.99 ± 53.57) contrasted to the control (232.62 ± 21.00). The activity of SOD gradually enhanced accompanied with the elevation of fluoride, and the activity of cells exposed to fluoride 48 h dramatically enhanced in 0.750 mmol/L group(1 298.66 ± 248.53)?1.250 mmol/L group (1 664.70 ± 277.68) compared to the control (741.67 ± 265.56). The activity of CAT in cells exposed to 0.750 mmol /L obviously elevated at 24 h (6.86 ± 1.08)?48 h(4.93 ± 0.16)? 72 h(4.83 ± 0.35) compared to the control. The mRNA level of bcl-2 markedly descended in 0.250 mmol/L group(0.80 ± 0.15), 0.375 mmol/L group(0.71 ± 0.16) compared to the control group(1.19 ± 0.03). Conclusions Oxidative stress may play a role in the development of renal injury induced by NaF. The lower level of Bcl-2 may descend the ability of anti-oxidative stress in renal tubule induced by NaF.

关 键 词：氟化物 氧化性应激 BCL-2基因 细胞培养 氟中毒 

分 类 号：R114[医药卫生—卫生毒理学]