牛磺酸对草酸和草酸钙晶体诱导的肾小管上皮细胞损伤的保护作用  

Protective effect of taurine on HK-2 cells exposed to oxalate and calcium oxalate monohydrate crystalin vivo

作　　者：黎承杨[1] 戚剑烽[1] 王翔[1] 陶芝伟[1] 邓耀良[1] 

机构地区：[1]广西医科大学第一附属医院泌尿外科,南宁530021

出　　处：《中华泌尿外科杂志》2013年第4期263-267,共5页Chinese Journal of Urology

基　　金：国家自然科学基金（0860280）

摘　　要：目的探讨牛磺酸在体外细胞培养中对草酸和草酸钙晶体诱导的肾小管上皮细胞损伤的保护作用及机制。方法将体外培养的人远端肾小管上皮细胞（HK-2）随机分为5组：第1组，单纯HK-2细胞；第2组，HK-2细胞中加入草酸和-水草酸钙（COM）；第3组，HK-2细胞中加入草酸、COM和牛磺酸；第4组，HK-2细胞中加入草酸、COM和夹竹桃麻素；第5组，HK-2细胞中加入草酸、COM和过氧化氢酶。培养6h后收集各组细胞上清液检测乳酸脱氢酶（LDH）、过氧化氢（H2O2）、8-异前列腺素（8-IP）含量及单核细胞趋化蛋白质-1（MCP-1）蛋白水平，RT—PCR法检测细胞内MCP-1mRNA、P47phoxmRNA表达水平；MTT法测定培养24h后的细胞存活率。结果培养24h后MTT法检测第1组细胞的吸光度A值为0．996±0．044，第2组为0．626±0．011，两组比较差异有统计学意义（P〈0．05）。培养6h后第1组细胞上清液H2O2、8-IP、LDH含量分别为（18．68±0．70）mmol／L、（12．72±1．69）ng／ml、（194．05±7．91）U／L，第2组分别为（53．27±1．88）mmol／L、（57．53±3．11）ng／ml、（484．34±29．44）U／L，两组比较差异均有统计学意义（P〈0．05）。第1组MCP-1蛋白水平为（17．32±2．32）pg／ml，第2组为（69．21±3．39）pg／ml，两组比较差异有统计学意义（P〈0．05）。第1组与第2组细胞的MCP．1mRNA、P47phoxmRNA表达水平倍增比值分别为1／3．51和1／1．38，两组比较差异均有统计学意义（P〈0．05）。MTT法检测第3、4、5组细胞的吸光度A值分别为0．748±0．033、0．825±0．078、0．815±0．037，与第2组比较差异均有统计学意义（P〈0．05）。第3、4、5组细胞上清液H2O2、8-IP、LDH含量均明显少于第2组。第3、4、5组细胞上清液MCP-1蛋白水平分别为（60．13±1．83）、（28．53±1．41）、（56．44±1．13）pg／ml，与第2组比较差异均有统计学意义（P〈0．05）Objective To investigate the protective effect of taurine on HK-2 cells exposed to ox- alate （Ox） and calcium oxalate monohydrate crystal （COM） in vivo. Methods HK-2 cells, a proximal tubular epithelial cell line, were cultured. Five groups were divided in this study： control group （only HK- 2 ceils）; Ox and COM group （HK-2 cells ＋ Ox ＋ COM） ; Taurine group （HK-2 ceils ＋ Ox ＋ COM ＋ Tau- rine） ; Apocynin group （HK-2 cells ＋ Ox ＋ COM ＋ Apocynin） ; Catalase group （ HK-2 cells ＋ Ox ＋ COM ＋Catalase）. After 6 hrs, the cultures medias from each group were tested for LDH, H2 O2 , 8-isoprostane, and MCP-1 protein. Cellular expression of MCP-1 mRNA and P47phox mRNA were determined by reverse tran- scriptase-polymerase chain reaction. After 24 hrs, cells livability was investigated by MTT. Results Compared with the control, cells livability was reduced when exposed to Ox and COM （P 〈 0.05 ） , Treat- ment with Taurine, Apocynin and Catalase significantly increased the cells livability （P 〈 0.05）. Compared with the control, the expression of LDH, H2O2 , 8-isoprostane, and cellular expression of MCP-1 mRNA and P47phox mRNA were increased following exposure to Ox and COM （P〈0.01, P〈0.01, P〈0.01, P〈 0.01, P 〈0.05 ）. Treatment with Taurine, Apocynin and Catalase significantly reduced the expression of LDH, H2O2, 8-isoprostane, as well as the cellular expression of MCP-1 mRNA. Expression of P47phox mRNA in Taurine group was not reduced significantly （P 〉 0.05 ）. Conclusions This study showed that Taurine protected the HK-2 cells from oxidative injury exposed to Ox and COM by the pathway that may not be in relation to the inhibition of P47phox mRNA expression.

关 键 词：肾小管上皮细胞 草酸 一水草酸钙 氧化应激 牛磺酸 

分 类 号：R692[医药卫生—泌尿科学]