羟基酪醇保护造影剂诱导的肾脏损伤并抑制内质网应激

The protective effect of hydroxytyrosol on contrast-induced nephropathy and endoplasmic reticulum stress

目的 研究造影剂肾病大鼠肾脏中葡萄糖调节蛋白78（GRP78）及半胱氨酸天冬氨酸蛋白酶12（Caspase-12）的表达情况,探讨内质网应激在造影剂肾病发病中的作用及羟基酪醇的干预作用.方法 84只大鼠随机分为3组：对照组（n=28）、模型组（n=28）和羟基酪醇干预组（n=28）.分别于造模后12 h、24 h、48 h、72 h处死部分大鼠,检测各组大鼠血尿素氮（BUN）、血肌酐（Scr）;ELISA法检测血清中丙二醛（MDA）、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）的含量;HE染色观察肾脏病理改变;原位末端转移酶标记法（TUNEL）检测肾小管上皮细胞凋亡;实时荧光定量PCR法检测肾组织GRP78 mRNA及Caspase-12mRNA的表达;免疫组化和Western印迹法检测各组大鼠肾组织GRP78及Caspase-12蛋白的表达.结果 对照组大鼠肾组织无明显病理损伤,GRP78、Caspase-12在mRNA水平及蛋白水平均无明显表达.与对照组相比,模型组大鼠肾组织病理损伤明显,细胞凋亡严重,且以上指标均显著升高（P＜0.05）.羟基酪醇干预组大鼠肾组织病理损伤、细胞凋亡较模型组减轻,且GRP78、Caspase-12在mRNA水平及蛋白水平表达均较模型组明显降低（P＜0.05）,但仍高于对照组（P＜0.05）.结论 GRP78及Caspase-12介导的内质网应激可能参与大鼠造影剂肾病的发生发展;羟基酪醇在造影剂诱导的肾脏损伤中发挥保护作用,这可能与其减轻内质网应激有关.

Objective To investigate the expression of glucose-regulated protein 78（GRP78）and cysteine aspartic acid protease 12（Caspase-12） and evaluate the endoplasmic reticulum stress （ERS） in rats with contrast-induced nephropathy （CIN）,and observe the protective effects of hydroxytyrosol on CIN rats.Methods Eighty-four Wistar rats,（220±20） g,were randomly divided into control group,CIN group,hydroxytyrosol treated group （group C＋H）.At 12th,24th,48th,72th day after the rats model were established,BUN and Scr were detected.ELISA were used to detect the expression of methane dicarboxylic aldehyde （MDA）,superoxide dismutase （SOD） and glutathione peroxidase （GSH-Px）.HE staining were used to evaluate the pathological change of kidney.TUNEL were used to detect the apoptosis of tubular ceils.Real-time PCR were used to detect the expression of GRP78 mRNA and Caspase-12 mRNA in tubular cells.Immunohistochemistry and Western blotting were used to detect the expression of GRP78 and Caspase-12 protein in tubular cells.Results BUN,Scr,the mRNA and protein expression of GRP78,Caspase-12 in hydroxytyrosol treated group were higher than that in control group（P 〈 0.05）,but were significantly lower than that in CIN group （P 〈 0.05）.Pathological changes and the apoptosis of tubular cells in CIN group were more serious than that in hydroxytyrosol treated group （P 〈 0.05）.Conclusions Endoplasmic reticulum stress may be associated with contrast-induced nephropathy.Hydroxytyrosol can protect kidney from contrast medium via reducing the endoplasmic reticulum stress.