摘要
目的 研究姜黄素对小鼠肾脏缺血再灌注后细胞自噬及炎症反应的影响.方法 选择健康、雄性的C57BL/6小鼠40只,随机数字表法平均分为4组.假手术组小鼠仅接受中线开腹、双侧肾蒂游离及关腹操作;再灌注组、姜黄素组及3-甲基腺嘌呤组小鼠均建立肾脏再灌注模型,并于肾脏缺血同时经尾静脉分别注射生理盐水、姜黄素（10 mg/kg）以及姜黄素（10 mg/kg）＋自噬抑制剂3-甲基腺嘌呤（15 mg/kg）.再灌注6、24 h检测各组血清肌、尿素氮水平,观察肾脏组织形态学变化;实时定量逆转录聚合酶链（RT-PCR）反应及蛋白印迹法检测肾脏组织内自噬相关分子兔抗小鼠微管相关蛋白1轻链3（LC3）、Beelin 1、RAS相关蛋白7（Rab7）、溶酶体相关膜蛋白2（LAMP2）的mRNA及蛋白表达,同时采用RT-PCR以及酶联免疫吸附实验检测肾组织及血清内白细胞介素6（IL-6）、IL-17、肿瘤坏死因子α（TNF-α）以及IL-10表达水平.结果 与再灌注组相比,姜黄素组小鼠再灌注6及24 h血清肌酐、尿素氮水平明显降低（P＜0.01）,肾小管上皮细胞损伤情况明显改善（P＜0.01）;肾脏组织内LC3、Beclin 1、Rab7以及LAMP2的mRNA及蛋白表达明显增加,肾脏组织及血清内炎症细胞因子IL-6、IL-17、TNF-α的表达降低,而IL-10的表达升高（P＜0.01）.而与姜黄素组相比,3-甲基腺嘌呤组肾功能及肾组织损害加重,自噬相关分子表达减少,炎症细胞因子表达增强而抗炎症细胞因子表达降低（P＜0.01）.结论 姜黄素对小鼠肾脏再灌注损伤的保护作用可能与促进再灌注后细胞自噬进而减轻炎症反应有关.
Objective To investigate the effect of curcumin on autophagy and inflammatory response in mice subject to renal ischemia reperfusion.Method Forty male C57BL/6 mice were randomly divided into four groups （n =10 each）：sham-operated （SO） group （Abdominal incision was made to expose the kidneys,bilateral renal pedicle dissociated and the abdomen sewed）,ischemia reperfusion （IR） group,curcumin （CM） group （given CM 10 mg/kg） and 3-methyladenine （3-MA） group （given 3-MA 15 mg/kg）.Six and 24 h after reperfusion,renal function was tested by determining the serum creatinine （Scr） and blood urea nitrogen （BUN） levels,and the morphological changes in the kidney tissue were observed.The expression of LC3,Beclin 1,Rab7 and LAMP2 in kidney tissue was detected by fluorescence quantitative RT-PCR and Western blotting.Fluorescence quantitative reverse transcription PCR and enzyme-linked immunosorbent assay were used to examine the expression of IL-6,IL-10,IL-17 and TNF-α in kidney tissue and serum.Result As compared with IR group and 3-MA group,the Scr and BUN levels in the CM group were significantly decreased （P〈0.01）,and the renal morphological changes were improved significantly （P〈0.01）.The mRNA and protein expression of LC3,Beclin 1,Rab7 and LAMP2 was significantly increased in kidney tissue,and the expression of IL-6,IL-17 and TNF-α was reduced,while IL-10 was increased （P〈 0.01） in the CM group as compared with IR group and 3-MA group.Conclusion Curcumin possesses a protective effect against renal ischemia reperfusion injury in mice,which is probably mediated by promoting autophagy and subsequently inhibiting inflammatory response.
出处
《中华器官移植杂志》
CAS
CSCD
2015年第12期724-730,共7页
Chinese Journal of Organ Transplantation
基金
基金项目：国家自然科学基金（81270550）
