时钟基因Bmal1调控过氧化物酶体增殖物激活受体α/氧化物酶体增殖物活化受体协同刺激因子1α通路减轻移植肾缺血再灌注损伤的研究  

Clock gene Bmal1 alleviate ischemia-reperfusion injury in transplanted kidney by regulate peroxisome proliferator activated receptorα/peroxisome proliferator activated receptor coactivator-1αpathway

作　　者：李维[1] 夏中元[1] 李文远 熊永红 冷燕[1] 赵胜[2] 程帆[2] Li Wei;Xia Zhongyuan;Li Wenyuan;Xiong Yonghong;Leng Yan;Zhao Sheng;Cheng Fan(Department of Anesthesiology,Renmin Hospital of Wuhan University,Wuhan 430060,China;Department of Urology,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院麻醉科,430060 [2]武汉大学人民医院泌尿外科,430060

出　　处：《中华实验外科杂志》2019年第11期2078-2081,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81671891)。

摘　　要：目的探讨时钟基因Bmal1调控过氧化物酶体增殖物激活受体α(PPARα)/氧化物酶体增殖物活化受体协同刺激因子1α(PGC-1α)通路对移植肾(TK)缺血再灌注损伤(IRI)的作用。方法体内实验验证Bmal13条短发卡RNA(shRNA)序列敲低Bmal1的效果,选取干预效果最好的第二条序列用于后续实验。将24只雄性SD大鼠随机分为4组:假手术组,TK组,TK+Bmal1 shRNA组,TK+Bmal1 shRNA+Fenofibrate(PPARα激动剂)组。各组给予相应的干预,血清检测肌酐(Cr)和尿素氮(BUN)含量;苏木精-伊红(HE)染色观察肾脏组织病理学变化;蛋白质印迹法(Western blot)检测肾组织Bmal1、PPARα、PGC-1α、B细胞淋巴瘤/白血病-2(bcl-2)/bcl-2相关X蛋白(bax)蛋白表达量;比色法检测肾组织三磷酸腺苷(ATP)生成量。使用SPSS 17.0软件进行统计分析。结果与假手术组(73.5±6.7、8.2±0.7、2.16±0.20、3.15±0.29、6.28±0.42、2.09±0.22、29.33±3.14)比较,TK组血清Cr(211±17.4)、BUN(44±3.8)含量及肾组织损伤程度均增加(t=18.063、22.695,P<0.05),而肾组织Bmal1(1.63±0.17)、PPARα(2.52±0.20)、PGC-1α(4.65±0.37)、bcl-2/bax(1.22±0.14)及ATP(15.34±1.02)生成量降低(t=4.946、4.450、7.133、8.172、10.380,P<0.05);与TK组比较,TK+Bmal1 shRNA组血清Cr(307.0±28.9)、BUN(53.0±4.2)含量及肾组织损伤程度均增加(t=6.971、3.892,P<0.05),而肾组织Bmal1(1.30±0.11)、PPARα(2.01±0.23)、PGC-1α(3.86±0.20)、bcl-2/bax(0.76±0.06)及ATP(10.15±0.96)生成量进一步降低(t=3.992、4.099、4.601、7.398、9.076,P<0.05);与TK+Bmal1 shRNA组比较,TK+Bmal1 shRNA+Fenofibrate组血清Cr(241.3±20.2)、BUN(47.0±4.5)含量及肾组织损伤程度均降低(t=4.564、2.388,P<0.05),而肾组织Bmal1(1.55±0.10)、PPARα(2.48±0.17)、PGC-1α(4.36±0.25)、bcl-2/bax(1.10±0.11)及ATP(14.71±1.21)生成量增加(t=4.119、4.025、3.826、6.647、29.572,P<0.05)。结论时钟基因Bmal1可通过活化PPARα/PGC-1α通路减轻移植肾缺血再灌注损伤。Objective This study was to investigate the effect of clock gene Bmal1 on ischemia-reperfusion injury in transplant kidney(TK)and relationship with peroxisome proliferator activated receptorα(PPARα)/peroxisome proliferator activated receptor coactivator-1α(PGC-1α)pathway.Methods Twenty-four male Sprague-Dawley rats were randomly divided into 4 groups:sham operation group,TK group,Bmal1 short hairpin RNA(shRNA)group,Bmal1 shRNA+fenofibrate group.Each group was given appropriate intervention and took specimens.The levels of serum creatinine(Cr)and urea nitrogen(BUN)were measured.The histopathological changes of kidneys in each group were estimated by hematoxylin and eosin(HE)staining.Western blotting was used to detect the expression of Bmal1,PPARα,PGC-1α,bcl-2/bax in kidney tissue.The amount of adenosine triphosphate(ATP)produced was determined by using colorimetric kit.SPSS 17.0 software was used for statistical analysis.The measurement data were expressed as mean±standard deviation.The comparison between two groups was analyzed with Student’s t test.The comparison among three or more groups was performed by one-way ANOVA test.Results Compared with the sham operation group(73.5±6.7,8.2±0.7,2.16±0.20,3.15±0.29,6.28±0.42,2.09±0.22,29.33±3.14),the serum Cr(211.0±17.4)and BUN levels(44.0±3.8)and renal tissue damage were increased(t=18.063,22.695,P<0.05),while the expression of Bmal1(1.63±0.17),PPARα(2.52±0.20),PGC-1α(4.65±0.37),bcl-2/bax(1.22±0.14)and ATP production(15.34±1.02)were decreased(t=4.946,4.450,7.133,8.172,10.380,P<0.05)in the TK group.Compared with the TK group,Bmal1 shRNA group showed increased Cr(307.0±28.9)and BUN(53.0±4.2)levels and renal tissue damage was increased(t=6.971,3.892,P<0.05),but decreased expression of Bmal1(1.30±0.11),PPARα(2.01±0.23),PGC-1α(3.86±0.20),bcl-2/bax(0.76±0.06)and ATP production(10.15±0.96)(t=3.992,4.099,4.601,7.398,9.076,P<0.05).Compared with Bmal1 shRNA group,the serum Cr(241.3±20.2)and BUN levels(47.0±4.5)and renal tissue damage were dec

关 键 词：移植肾 肾脏缺血再灌注损伤 能量代谢 BMAL1 过氧化物酶体增殖物激活受体α/氧化物酶体增殖物活化受体协同刺激因子1α 

分 类 号：R[医药卫生]