Abstract：
To explore the change of clock gene rhythm under renal denervation (RDN) and its effect on renal function and oxidative stress during renal ischemia-reperfusion (IR) injury.
C57/BL6 mice were randomly divided into 4 groups at daytime 7 A M (zeitgeber time [ZT] 0) or at nighttime 7 P M (ZT12) in respectively: Sham (S) group, RDN group, IR group and RDN + IR (DIR) group. Renal pathological and functional changes were assessed by H&E staining, and serum creatinine, urea nitrogen and neutrophil gelatinase-associated lipocalin levels. Renal oxidative stress was detected by SOD and MDA levels, and renal inflammation was measured by IL-6, IL-17 A F and TNF-ɑ levels. BMAL1, CLOCK, Nrf2 and HO-1 mRNA and protein expressions were tested by qPCR and Western Blot.
Compared with S groups, the rhythm of BMAL1, CLOCK and Nrf2 genes in the kidney were disordered in RDN groups, while renal pathological and functional indexes did not change significantly. Compared with IR groups, renal pathological and functional indexes were significantly higher in the DIR groups, as well as oxidative stress and inflammation in renal tissues. The nocturnal IR injury in the RDN kidney was the worst while the BMAL1, Nrf2 and HO-1 expressions were the highest. In DIR groups, renal injury was aggravated after the Brusatol treatment, but there was no significant improvement after the t-BHQ treatment at night, which might be consistent with the changes of Nrf2 and HO-1 protein expressions.
RDN lead to the disruption of BMAL1-mediated Nrf2 rhythm accumulation in the kidney, which reduced the renal ability to resist oxidative stress and inflammation, due to the impaired effect of activating Nrf2/ARE pathway in renal IR injury at nighttime.
C57/BL6 mice were randomly divided into 4 groups at daytime 7 A M (zeitgeber time [ZT] 0) or at nighttime 7 P M (ZT12) in respectively: Sham (S) group, RDN group, IR group and RDN + IR (DIR) group. Renal pathological and functional changes were assessed by H&E staining, and serum creatinine, urea nitrogen and neutrophil gelatinase-associated lipocalin levels. Renal oxidative stress was detected by SOD and MDA levels, and renal inflammation was measured by IL-6, IL-17 A F and TNF-ɑ levels. BMAL1, CLOCK, Nrf2 and HO-1 mRNA and protein expressions were tested by qPCR and Western Blot.
Compared with S groups, the rhythm of BMAL1, CLOCK and Nrf2 genes in the kidney were disordered in RDN groups, while renal pathological and functional indexes did not change significantly. Compared with IR groups, renal pathological and functional indexes were significantly higher in the DIR groups, as well as oxidative stress and inflammation in renal tissues. The nocturnal IR injury in the RDN kidney was the worst while the BMAL1, Nrf2 and HO-1 expressions were the highest. In DIR groups, renal injury was aggravated after the Brusatol treatment, but there was no significant improvement after the t-BHQ treatment at night, which might be consistent with the changes of Nrf2 and HO-1 protein expressions.
RDN lead to the disruption of BMAL1-mediated Nrf2 rhythm accumulation in the kidney, which reduced the renal ability to resist oxidative stress and inflammation, due to the impaired effect of activating Nrf2/ARE pathway in renal IR injury at nighttime.
摘要：
目的：探讨肾缺血再灌注损伤时去神经(RDN)时钟基因节律的变化及其对肾功能和氧化应激的影响。
方法：C57/BL6小鼠随机分为4组，分别在白天7AM（zitgeber时间[ZT]0）或夜间7PM（ZT12）：假（S）组，RDN组,IR组和RDN+IR(DIR)组。通过H&E染色评估肾脏病理和功能改变，和血清肌酐,尿素氮和中性粒细胞明胶酶相关脂质运载蛋白水平。肾脏氧化应激检测SOD和MDA含量,检测IL-6、IL-17AF和TNF-α水平。BMAL1,时钟,通过qPCR和WesternBlot检测Nrf2和HO-1的mRNA和蛋白表达。
结果：与S组相比，RDN组肾脏BMAL1、CLOCK和Nrf2基因节律紊乱,而肾脏病理和功能指标无明显变化。与IR组相比，肾脏病理和功能指标在DIR组明显增高,以及肾组织中的氧化应激和炎症。RDN肾夜间IR损伤最严重,BMAL1、Nrf2和HO-1表达最高。在DIR组中，Brusatol治疗后肾损伤加重，但是在夜间t-BHQ治疗后没有显着改善，这可能与Nrf2和HO-1蛋白表达的变化相一致。
结论：RDN导致肾脏中BMAL1介导的Nrf2节律积累的破坏，降低了肾脏抵抗氧化应激和炎症的能力，由于激活Nrf2/ARE通路在夜间肾IR损伤中的作用受损。
方法：C57/BL6小鼠随机分为4组，分别在白天7AM（zitgeber时间[ZT]0）或夜间7PM（ZT12）：假（S）组，RDN组,IR组和RDN+IR(DIR)组。通过H&E染色评估肾脏病理和功能改变，和血清肌酐,尿素氮和中性粒细胞明胶酶相关脂质运载蛋白水平。肾脏氧化应激检测SOD和MDA含量,检测IL-6、IL-17AF和TNF-α水平。BMAL1,时钟,通过qPCR和WesternBlot检测Nrf2和HO-1的mRNA和蛋白表达。
结果：与S组相比，RDN组肾脏BMAL1、CLOCK和Nrf2基因节律紊乱,而肾脏病理和功能指标无明显变化。与IR组相比，肾脏病理和功能指标在DIR组明显增高,以及肾组织中的氧化应激和炎症。RDN肾夜间IR损伤最严重,BMAL1、Nrf2和HO-1表达最高。在DIR组中，Brusatol治疗后肾损伤加重，但是在夜间t-BHQ治疗后没有显着改善，这可能与Nrf2和HO-1蛋白表达的变化相一致。
结论：RDN导致肾脏中BMAL1介导的Nrf2节律积累的破坏，降低了肾脏抵抗氧化应激和炎症的能力，由于激活Nrf2/ARE通路在夜间肾IR损伤中的作用受损。
